ResearchArticle Heuchera Creme Brulee and Mahogany...
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Research Article Heuchera Creme Brulee and Mahogany Medicinal Value under Water Stress and Oligosaccharide (COS) Treatment Hosam O. Elansary , 1,2,3 Amal M. E. Abdel-Hamid, 4 Eman A. Mahmoud, 5 Fahed A. Al-Mana, 1 Diaa O. El-Ansary, 6 and Tarek K. Zin El-Abedin 7 1 Plant Production Department, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia 2 Floriculture, Ornamental Horticulture and Garden Design Department, Faculty of Agriculture (El-Shatby), Alexandria University, Alexandria, Egypt 3 Department of Geography, Environmental Management and Energy Studies, University of Johannesburg, APK Campus, 2006, South Africa 4 Department of Biological and Geological Sciences, Faculty of Education, Ain Shams University, Cairo, Egypt 5 Department of Food Industries, Damietta University, Damietta, Egypt 6 Precision Agriculture Laboratory, Department of Pomology, Faculty of Agriculture (El-Shatby), Alexandria University, Alexandria, Egypt 7 Department of Agricultural Engineering, College of Food and Agriculture Sciences, King Saud University, Riyadh, Saudi Arabia Correspondence should be addressed to Hosam O. Elansary; [email protected] Received 24 October 2018; Accepted 28 January 2019; Published 17 February 2019 Academic Editor: Letizia Angiolella Copyright © 2019 Hosam O. Elansary et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Food borne pathogens cause serious human illnesses and diseases and their control using natural bioactive compounds becomes essential for the progress of agricultural and food industries. Developing novel tools to enhance the medicinal values of traditional horticultural medicinal crops is one of the promising methods for achieving food borne pathogens control. In this study, oligosaccharide water solutions were applied to Heuchera Creme Brulee and Mahogany subjected to a normal irrigation interval (2 days) or to prolonged irrigation intervals (6 days) for 6 weeks. Plant morphological, physiological, and metabolic markers associated with the bioactivity of leaf extracts against selected microbes. Oligosaccharide-treated plants showed significant increases in all morphological parameters during normal and prolonged irrigation intervals as compared to those of the controls. Morphological improvement associated with a significant increase in chlorophyll, carbohydrates, proline, K, Ca, phenols, and free and total ascorbate and antioxidants. Superoxide dismutase, catalase, and ascorbate peroxidase activities were higher, while H 2 O 2 accumulated to a lower extent in oligosaccharide-treated plants. ese morphological and metabolic changes associated with increased antibacterial and antifungal activities of leaf extracts and their activities were comparable to antibiotics and antifungal agents (minimum inhibitory concentrations values were 0.5 -0.20 mg −1 mL for bacteria and 0.08 -0.20 mg −1 mL for fungi in Mahogany). e application of oligosaccharide and/or water stress might be of great value for producing natural bioactive compounds for food borne pathogens control. 1. Introduction Food borne illnesses such as diarrheal and emetic symptoms are of great importance in the agricultural industry including milk processing worldwide [1, 2]. ese illnesses are caused by several microbes such as the bacterial including Bacillus cereus [3] and Listeria monocytogenes [4, 5] and the severity of the relevant diseases may cause human death. Fungi cause massive agricultural losses and threatens human food storage facilities and produce mycotoxins that cause cancer diseases and neurological disorders. ese fungi include Aspergillus niger that causes black mold on several horticultural crops and Aspergillus ochraceus that contaminate human foods [6–8] and both species developed resistance to antifungal Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2019, Article ID 4242359, 13 pages https://doi.org/10.1155/2019/4242359
Transcript of ResearchArticle Heuchera Creme Brulee and Mahogany...
Research ArticleHeuchera Creme Brulee and Mahogany Medicinal Value underWater Stress and Oligosaccharide (COS) Treatment
HosamO Elansary 123 Amal M E Abdel-Hamid4 Eman A Mahmoud5
Fahed A Al-Mana1 Diaa O El-Ansary6 and Tarek K Zin El-Abedin7
1Plant Production Department College of Food and Agriculture Sciences King Saud University PO Box 2460Riyadh 11451 Saudi Arabia
2Floriculture Ornamental Horticulture and Garden Design Department Faculty of Agriculture (El-Shatby)Alexandria University Alexandria Egypt
3Department of Geography Environmental Management and Energy Studies University of JohannesburgAPK Campus 2006 South Africa
4Department of Biological and Geological Sciences Faculty of Education Ain Shams University Cairo Egypt5Department of Food Industries Damietta University Damietta Egypt6Precision Agriculture Laboratory Department of Pomology Faculty of Agriculture (El-Shatby)Alexandria University Alexandria Egypt
7Department of Agricultural Engineering College of Food and Agriculture Sciences King Saud University Riyadh Saudi Arabia
Correspondence should be addressed to Hosam O Elansary helansaryksuedusa
Received 24 October 2018 Accepted 28 January 2019 Published 17 February 2019
Academic Editor Letizia Angiolella
Copyright copy 2019 Hosam O Elansary et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited
Food borne pathogens cause serious human illnesses and diseases and their control using natural bioactive compounds becomesessential for the progress of agricultural and food industries Developing novel tools to enhance the medicinal values oftraditional horticultural medicinal crops is one of the promising methods for achieving food borne pathogens control In thisstudy oligosaccharide water solutions were applied to Heuchera Creme Brulee and Mahogany subjected to a normal irrigationinterval (2 days) or to prolonged irrigation intervals (6 days) for 6 weeks Plant morphological physiological and metabolicmarkers associatedwith the bioactivity of leaf extracts against selectedmicrobes Oligosaccharide-treated plants showed significantincreases in all morphological parameters during normal and prolonged irrigation intervals as compared to those of the controlsMorphological improvement associated with a significant increase in chlorophyll carbohydrates proline K Ca phenols andfree and total ascorbate and antioxidants Superoxide dismutase catalase and ascorbate peroxidase activities were higher whileH2O2accumulated to a lower extent in oligosaccharide-treated plants These morphological and metabolic changes associated
with increased antibacterial and antifungal activities of leaf extracts and their activities were comparable to antibiotics andantifungal agents (minimum inhibitory concentrations values were 05 -020 mgminus1mL for bacteria and 008 -020 mgminus1mL forfungi in Mahogany) The application of oligosaccharide andor water stress might be of great value for producing natural bioactivecompounds for food borne pathogens control
1 Introduction
Food borne illnesses such as diarrheal and emetic symptomsare of great importance in the agricultural industry includingmilk processing worldwide [1 2] These illnesses are causedby several microbes such as the bacterial including Bacilluscereus [3] and Listeria monocytogenes [4 5] and the severity
of the relevant diseases may cause human death Fungi causemassive agricultural losses and threatens human food storagefacilities and produce mycotoxins that cause cancer diseasesand neurological disorders These fungi include Aspergillusniger that causes black mold on several horticultural cropsand Aspergillus ochraceus that contaminate human foods[6ndash8] and both species developed resistance to antifungal
HindawiEvidence-Based Complementary and Alternative MedicineVolume 2019 Article ID 4242359 13 pageshttpsdoiorg10115520194242359
2 Evidence-Based Complementary and Alternative Medicine
agents [9] In the same trend food borne bacteria developedsignificant resistance to antibiotics [10] which steamed thesearch for natural alternatives that have more ability tocontrol food borne pathogens To reduce the losses in thefood industry and to maintain the food security the useof synthetic food preservatives was introduced to the foodindustry although these preservatives had severe side effectson the human health on the long run [11] These conditionsoriented the search for natural bioactive compounds thathave the capabilities to control food borne pathogens
Horticultural crops tend to produce secondary metabo-lites during stress conditions such as water stressWater stressis one of the major limiting factors for agricultural industryespecially in view of the rapidly increasing world populationglobal climate change and the increasing worldwide indus-trial demand for water [12] Water stress may have severalmorphological (eg leaf number and leaf area) physiological(eg carbohydrate and ion composition) metabolic (egSOD activity and composition) and molecular (eg freeradical scavenging gene products) effects on plants leadingto reduced yields as well as increased accumulation of severalcompounds Plant metabolic responses to water stress mayinclude the accumulation of carbohydrates [13] increasedsynthesis of specific proteins increased stress related nutrientuptake (eg K) and accumulation of specific antioxidantssuch as the phenolic compounds and others that neutralizereactive oxygen species (ROS) [14ndash17]
Efforts to develop novel tools to enable horticultural cropsto cope with water stress on plants are a growing concernworldwide such as the use of biochar [18] 120573-aminobutyricacid [19] trinexapac-ethyl [20] seaweed extracts [21]nanoparticles [22] and oligosaccharide Oligosaccharide isa biostimulant produced commercially by subjecting chitinto high temperature followed by deacetylation using alka-line conditions to remove proteins and calcium [23 24]Oligosaccharides may be formulated as a solution or aswater-soluble powder They are widely used as plant elicitorsof the production of secondary metabolites [25] particu-larly polyphenols [26] Oligosaccharides have also strongantimicrobial activities and may stimulate the growth ofbeneficiary microbes [27] Additionally several studies sug-gest that it may improve crop yield [23] and enhancestress tolerance [28] However little is known regarding themechanism whereby oligosaccharides enhance water stresstolerance and effect secondary metabolites in horticulturalcrops
Saxifragaceae includes 30 genera of herbaceous peren-nials such as Heuchera which are known to be geneticallydiverse due to hybridization [29] Heuchera contains about50 species One of these is Heuchera which accommodatesperennial herbaceous ornamental shade plants widely usedin North America Europe North Africa and South Asia[30] Dozens of colored hybrid cultivars varying in leaf andflower color have been recently introduced in the marketInterestingly although native people of Europe have usedHeuchera and other genera of Saxifragaceae as traditionalmedicinal plants [31] for centuries the medicinal propertiesresponses of this species to oligosaccharide elicitors underwater stress have not been investigated
In the present study our objective was to explore thepossible effects of oligosaccharides onHeuchera grown undernormal and prolonged irrigation intervals by using morpho-logical physiological and metabolic markers We hypothe-sized that stress conditions and oligosaccharides treatmentmay enhance antimicrobial properties of Heuchera plantsThe information obtained from this study will contribute toour understanding of oligosaccharides andor water stressaction in plant metabolic responses that may help in thediscovery and use of natural bioactive compounds controlfood spoilage microorganisms
2 Material and Methods
21 Plant Material and Treatments Young plants 10 cmhigh of Heuchera cultivars Creme Brulee and Mahoganywere obtained from local commercial nurseries on January7th 2017 and 2018 Plants were grown in a polyethylene-covered greenhouse located on the Alexandria-Cairo desertroad Egypt All plants were identified by Hosam Elansaryand registered at the Faculty of Agriculture AlexandriaUniversity prior to transplanting onto 21 L pots containinga mixture of brown peat and perlite (31 ww) supplementedwith Crystalon (20 N 20 P 20 K 2 gL media) Plantswere grown for three weeks under temperatures rangingbetween 151∘C and 275∘C relative humidity between 58and 67 photosynthetically active radiation around 1000mminus2 at 1200 pm and daily watering of 38-50mLplant Plantswere divided into two groups one of which was watered at2-day intervals (2DWI) while the other was watered at 6-dayintervals (6DWI) for 6weeks Oligosaccharide (deacetylationgt 95 MW501486 gmol powder Aldebeiky Group CoCairo Egypt) water solution was sprayed at concentrations of50 200 or 500 ppm until drop off 2 weeks prior to extendingthe watering interval untreated plants were considered asthe control treatment The experiment was laid out in asplit-plot design Irrigation intervals were considered as themain plot and oligosaccharides treatments the subplot Plantswere grouped into three blocksrepetitions (n=3) containing5 replicates per treatment for a total of 40 plants per cultivarper season in Randomized Complete Block Design (RCBD)
22 Morphological and Physiological Parameters Plants wereharvested after 6 weeks of stress treatment At that pointplant height and leaf number were registered Leaf area wascalculated immediately using a scanner and the AutoCADprogram Total dry weight was determined by drying cleanedplants to constant weight in an oven at 70∘C Total carbohy-drates K+ Ca2+ and proline were determined in plant leavesat the end of the experiment Following freeze-drying ofsamples they were ground and sieved and then kept at -20∘Cuntil further analysis Total carbohydrates were quantifiedafter Dubios et al [32] and expressed on a percent basis Onegram of frozen leaves was used to obtain the cell sap thena dilution (1100 vv) was used for the determination of K+andCa2+ concentrations using an inductively coupled plasmaspectrophotometer [33] Proline leaf content was determinedin the Department of Plant Production King Saud Universityusing a spectrophotometer at 520 nm [34 35]
Evidence-Based Complementary and Alternative Medicine 3
23 Antioxidants Chlorophyll Phenols andEnzymeActivitiesAir dried leaves were ground into fine powder 025 g of thisfrom each sample was dissolved into 3 mL methanol (99)while stirring on a magnetic agitator at low speed in thedark for 24 h at room temperature Methanolic extracts werecentrifuged for 5 min under cooling at 10000 RPM (7000 timesg) the supernatant (sim27mL)was dried in a rotary evaporatorto produce a semisolid extract which was stored for laterantioxidant analysis Antioxidant activities of all sampleswere determined in the Department of Plant ProducitonKing Saud University using the 221015840-diphenylpicrylhydrazyl(DPPH) and 120573-carotene-linoleic acid methods which mea-sure OHminus scavenging activities according to Elansary et al[21] For the DPPH method samples were incubated for 30min after which absorbance was measured at 517 nm Forthe 120573-carotene-linoleic acid assay absorbance was measuredat 470 nm The sample concentration required to scavenge50 of DPPH 120573-carotene-linoleic acid (IC
50in 120583gmL)
was determined by plotting the inhibition percentage againstextract concentration Butylated hydroxytoluene (BHT) wasused as a positive control and experiments were repeatedtwice in triplicate Total phenolic content in methanolicleaf extracts were performed using the Folin-Ciocalteaucolorimetric method using gallic acid as the reference andexpressing the results as gallic acid equivalents (mg GAEgminus1 ext) [36 37] Total chlorophyll content was quantified infresh leaves according to Moran and Porath [38]
Ground-frozen leaves were used to quantify total andfree ascorbate after Elansary et al [21] Briefly 05 g ofground-frozen leaf tissues were homogenized in 8 mL cooledtrichloroacetic acid (TCA 5 wv) next the mixture wascentrifuged for 10 min (10000 times g) at 4∘C The supernatantwas incubated with a mixture of PBS (200 mM pH 74)and dithiothreitol (DTT 15 mM) for 50 min excess DTTwas removed by adding N-ethylmaleimide (NEM 200 120583L05 wv) The solution was then mixed with TCA (1 mL10 wv) o-phosphoric acid (800 120583L 42 wv) and 22-dipyridyl in 70 (vv) ethanol (800 120583L 65 mM) and iron(III)chloride (400 120583L 3 wv) and incubated for 1 h at 42∘CAbsorbance by the mixture was measured at 525 nm Freeascorbate was determined using the same procedure exceptDTT and NEM were replaced with 400 120583L deionized waterwhile free and total ascorbate contents were determined usingstandard curves
Catalase (CAT) ascorbate peroxidase (APX) and super-oxide dismutase (SOD) activities as well as H
2O2accumula-
tion were quantified in leaves tissues following Elansary et al[21]
24Microorganisms andMedicinal Properties Themedicinalproperties of methanolic leaf extracts were studied againstselected pathogenic bacteria and fungi The selected bacteriawere Listeria monocytogenes (clinical isolate) Bacillus cereus(ATCC 14579) Staphylococcus aureus (ATCC 6538) Micro-coccus flavus (ATCC 10240) Pseudomonas aeruginosa (ATCC27853) and Escherichia coli (ATCC 35210)The selected fungiwere Aspergillus niger (ATCC 6275) A ochraceus (ATCC12066) A flavus (ATCC 9643) Penicillium ochrochloron(ATCC 48663) and Candida albicans (ATCC 12066) The
microdilution method [39] was used to determine theantibacterial and antifungal activities In the antibacterialassay the minimum inhibitory bactericidal concentration(MIC) was defined as the lowest concentration resultingin growth stop of the bacteria at the binocular level Theminimum bactericidal concentration (MBC) was definedas the lowest concentration resulting in killing 995 ofthe original inoculum Also the MBC was determined byserial subcultivation of the bacterial using 01-02 mgmL ofbacterial solution added to 100 120583L of TSB and incubatedfor one day In the antifungal activity assay the minimuminhibitory concentration (MIC) was defined as the lowestconcentration inhibiting the fungal growth at the binocularlevel while the minimum fungicidal concentration (MFC)was determined using subcultivations of the fungi (01-40mgmL) and was defined as the concentration killing 995of the original inoculum Experiments were performed twiceand negative controls (5 DMSO) as well as positive con-trols [antibacterial assay streptomycin and ampicillin 001-10 mgmL antifungal Fluconazole (FLZ) and ketoconazole(KLZ)] were used Experiments were repeated twice
25 Statistical Analyses The data obtained during the twogrowing seasons in 2017 and 2018 were expressed as meansand Least Significant Difference (LSD) was determined usingthe one way ANOVA test in SPSS (PASWVer 21) at P le 005
3 Results
31 Morphological and Physiological Responses to IrrigationIntervals and Oligosaccharide Increasing watering intervalsfrom 2 to 6 days significantly reduced morphological param-eters in bothHeuchera cultivars tested including leaf numberleaf area plant dry weight and plant height (Table 1) Inter-estingly under the normal irrigation interval (2DWI) theapplication of the oligosaccharide at 50 and 200 ppm signifi-cantly increased leaf number and area plant dry weight andplant height in both cultivars treated plants in both seasonscompared to untreated plants Further under prolongedirrigation interval (6DWI) there were significant increasesin both Creme Brulee and Mahogany in all morphologicalparameters measure in plants treated with oligosaccharide at50 and 200 ppm compared to oligosaccharide at 500 ppmand control treatment Prolonged irrigation interval (6DWI)significantly reduced total carbohydrates K Ca and prolinecontents in plants of both Creme Brulee and Mahoganycompared to the normal irrigation interval (2DWI) as shownin Table 2 Under 2DWI as well as 6DWI total carbohydratesK Ca and proline contents increased significantly in theleaves of oligosaccharides -treated plants at 50 and 200ppm compared to controls and 500 ppm oligosaccharidetreatment in both growing seasons
32 General Antioxidants Phenolics and ChlorophyllsExtension of irrigation interval from 2 to 6 days caused asignificant increase in DPPH free radical scavenging activityin both Heuchera cultivars (Table 3) The DPPH (IC
50) of
Creme Brulee plants decreased in the first season (2017)which indicates an increase in scavenging activity a similar
4 Evidence-Based Complementary and Alternative Medicine
Table1Eff
ecto
fwater
deficitandoligosaccharides
treatmento
nleafnu
mberleafareaplant
drywe
ightand
planth
eigh
tintwoHeucheracultivarsaft
ersix
weekso
ftreatmentinitia
tion
Values
aree
xpressed
asmeans
(plusmnsd)
Waterinterval
Oligosaccharides
treatment(pp
m)
Leafnu
mber(leafplantminus1)
Leafarea
(cm2plantminus1)
Plantd
rywe
ight
(gplantminus1)
Planth
eigh
t(cm
)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
156plusmn02blowast
152plusmn01b
6512plusmn151b
6482plusmn111b
112plusmn01b
112plusmn02b
291plusmn01b
288plusmn02b
50161plusmn01ab
161plusmn02a
6901plusmn
131a
6863plusmn145a
123plusmn01a
121plusmn02a
332plusmn01a
308plusmn03a
200
171plusmn
04a
172plusmn01a
7031plusmn
143a
6992plusmn151a
124plusmn01a
122plusmn02a
324plusmn02a
307plusmn02a
500
156plusmn01b
153plusmn02b
6392plusmn111b
6505plusmn
223b
113plusmn01b
112plusmn02b
301plusmn01b
294plusmn01b
6DWI
070plusmn01d
71plusmn02d
3031plusmn
103d
3111plusmn131d
55plusmn01d
56plusmn01d
173plusmn
03d
168plusmn01d
5086plusmn00cd
85plusmn03cd
3509plusmn151c
3581plusmn
112c
63plusmn01c
62plusmn01c
194plusmn01c
188plusmn01c
200
90plusmn01c
91plusmn02c
3613plusmn141c
3516plusmn175c
61plusmn
02c
62plusmn01c
195plusmn01c
190plusmn01c
500
73plusmn00d
72plusmn01d
3112plusmn131d
3061plusmn
141d
53plusmn01d
53plusmn01d
174plusmn02cd
172plusmn01d
2DWI
0Mahogany
132plusmn01b
128plusmn01b
5161plusmn
221b
5125plusmn101b
108plusmn03b
107plusmn01b
308plusmn03b
311plusmn04b
50144plusmn01a
142plusmn02a
5631plusmn
201a
5675plusmn112a
116plusmn01a
117plusmn02a
333plusmn01a
329plusmn03a
200
147plusmn03a
142plusmn03a
5731plusmn
103a
5781plusmn
121a
116plusmn01a
117plusmn01a
342plusmn04a
332plusmn03a
500
132plusmn01b
131plusmn01b
5103plusmn113b
5021plusmn
167b
108plusmn01b
108plusmn02b
314plusmn01b
307plusmn01b
6DWI
061plusmn
02e
62plusmn02e
2201plusmn
121d
2153plusmn152d
54plusmn01d
55plusmn01d
181plusmn01d
183plusmn03d
5074plusmn01d
71plusmn01d
2611plusmn131c
2683plusmn
113c
63plusmn01c
62plusmn01c
212plusmn03c
207plusmn01c
200
81plusmn
01 c
82plusmn02c
2713plusmn101c
2773plusmn111c
63plusmn01c
63plusmn01c
205plusmn02c
209plusmn01c
500
62plusmn01e
61plusmn
01e
2102plusmn115d
2215plusmn151d
55plusmn01d
54plusmn01d
183plusmn01d
187plusmn03d
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 5
Table2Eff
ectofirrigationintervalsand
oligosaccharidestre
atmentontotalcarbo
hydrateK
Caandprolinec
ontent
intheleaveso
ftwoHeucheracultivarsin
twosuccessiv
eseasonsV
alues
arem
eans
(plusmnsd)
Water
interval
Oligosaccharides
treatment
(ppm
)
Totalcarbo
hydrates
(DW)
K(m
ggminus1DW)
Ca(
mggminus1DW)
Proline(
mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
1345plusmn01blowast
1337plusmn01b
197plusmn01d
195plusmn05d
376plusmn005b
363plusmn004
b13
5plusmn005c
132plusmn001c
501433plusmn01a
1422plusmn01ab
248plusmn01b
239plusmn01b
412plusmn00a
411plusmn02a
144plusmn003b
141plusmn
000
cb200
1453plusmn02a
1467plusmn01a
258plusmn02b
249plusmn01b
415plusmn009a
409plusmn003a
147plusmn001b
144plusmn003b
500
1353plusmn01b
1349plusmn01b
199plusmn00d
194plusmn01d
385plusmn001b
379plusmn005b
137plusmn001c
135plusmn002c
6DWI
01219plusmn02c
1205plusmn01c
215plusmn01b
213plusmn03c
363plusmn006
b365plusmn006
b14
8plusmn002b
146plusmn001ab
501289plusmn01cb
1251plusmn
01c
273plusmn03a
268plusmn01a
405plusmn003a
397plusmn001a
156plusmn003a
153plusmn003a
200
1287plusmn01cb
1298plusmn01bc
279plusmn01a
277plusmn01a
419plusmn007a
411plusmn004
a15
8plusmn002a
155plusmn
002a
500
1232plusmn02c
1231plusmn01c
222plusmn02c
218plusmn02c
378plusmn006
b370plusmn005b
149plusmn003ab
147plusmn001ab
2DWI
0Mahogany
1532plusmn01b
a1505plusmn01b
213plusmn01c
217plusmn03c
361plusmn004
b367plusmn008b
142plusmn002c
138plusmn001c
501599plusmn01ab
1591plusmn
00a
264plusmn01b
261plusmn01b
397plusmn003a
394plusmn005a
155plusmn004
b14
6plusmn001b
200
1630plusmn01a
1613plusmn03a
268plusmn01b
266plusmn03b
406plusmn006
a401plusmn006
a15
8plusmn008b
149plusmn004
b500
1551plusmn
01b
1527plusmn03b
233plusmn03d
226plusmn01c
373plusmn004
b375plusmn003b
147plusmn001c
141plusmn
001c
6DWI
01441plusmn
02c
1412plusmn03c
255plusmn01b
247plusmn01b
358plusmn002b
368plusmn002b
156plusmn003b
151plusmn
004
b50
1545plusmn01b
1513plusmn01b
299plusmn01a
289plusmn03a
393plusmn001a
395plusmn001a
170plusmn001a
169plusmn002a
200
1555plusmn01b
1525plusmn05b
3 03plusmn01a
297plusmn02a
393plusmn001a
396plusmn004
a174plusmn003a
171plusmn
002a
500
1437plusmn01c
1424plusmn03c
264plusmn01a
258plusmn03b
363plusmn005b
367plusmn002b
159plusmn002b
154plusmn001b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
6 Evidence-Based Complementary and Alternative Medicine
Table3Antioxidant
activ
ityin
leafmethano
licextracts
totalpheno
licandtotalchlorop
hyllcontento
ftwo
HeucheracultivarsV
aluesa
remeans
oftriplicated
eterminationsplusmnsd
Water
interval
Oligosaccharides
treatment
(ppm
)
DPP
Hfre
eradical
scavenging
activ
ity(IC
50120583
gmlminus1
)
120573-C
arotene-lin
oleica
cid
assay
(IC50120583
gmlminus1
)
Totalp
heno
liccontent
(mgGAEgminus1)
Totalchlorop
hyllcontent
(mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
103plusmn001alowast
111plusmn
007a
112plusmn001a
115plusmn001a
104plusmn01c
97plusmn01c
065plusmn004
b063plusmn001bc
5093plusmn006
b99plusmn001b
103plusmn003b
104plusmn002b
109plusmn01b
104plusmn03b
069plusmn002a
067plusmn003a
200
91plusmn005b
99plusmn002b
103plusmn003b
107plusmn003b
108plusmn00b
105plusmn02b
070plusmn002a
068plusmn001a
500
96plusmn003a
104plusmn003a
112plusmn002a
115plusmn004
a104plusmn02c
100plusmn01c
068plusmn001ab
065plusmn002ab
6DWI
082plusmn004
c89plusmn005c
93plusmn001c
101plusmn003c
109plusmn04b
104plusmn02b
061plusmn002c
060plusmn003c
5063plusmn002d
68plusmn000
d74plusmn002d
82plusmn002d
116plusmn03a
112plusmn02a
065plusmn001b
064plusmn002b
200
62plusmn001d
68plusmn001d
72plusmn003d
82plusmn002d
116plusmn01a
113plusmn02a
065plusmn001b
065plusmn003ab
500
78plusmn003c
83plusmn001c
92plusmn004
c99plusmn001c
111plusmn
02b
105plusmn04b
062plusmn002c
061plusmn001c
2DWI
0Mahogany
89plusmn04a
96plusmn01a
101plusmn03a
107plusmn02a
123plusmn02c
117plusmn03c
071plusmn001b
070plusmn002b
5074plusmn002b
79plusmn001b
85plusmn001b
94plusmn003b
127plusmn03b
124plusmn01b
075plusmn002a
074plusmn001a
200
71plusmn001b
78plusmn004
b84plusmn000
b93plusmn003b
129plusmn04b
125plusmn02b
076plusmn001a
075plusmn001a
500
82plusmn002a
88plusmn003a
95plusmn003a
104plusmn002a
122plusmn01c
121plusmn03b
c072plusmn001b
070plusmn002b
6DWI
071plusmn003b
77plusmn002b
84plusmn001b
89plusmn003b
131plusmn01b
124plusmn02b
066plusmn002c
063plusmn001c
5054plusmn001c
57plusmn002c
66plusmn003c
72plusmn003c
138plusmn02a
132plusmn00a
070plusmn001c
068plusmn003c
200
48plusmn00 3c
55plusmn007c
62plusmn002c
71plusmn002c
139plusmn03a
134plusmn01a
071plusmn001a
068plusmn002a
500
68plusmn001b
74plusmn003b
79plusmn003b
82plusmn001b
134plusmn02ab
125plusmn01b
068plusmn002b
064plusmn003b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
different
basedon
LSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
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Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
2 Evidence-Based Complementary and Alternative Medicine
agents [9] In the same trend food borne bacteria developedsignificant resistance to antibiotics [10] which steamed thesearch for natural alternatives that have more ability tocontrol food borne pathogens To reduce the losses in thefood industry and to maintain the food security the useof synthetic food preservatives was introduced to the foodindustry although these preservatives had severe side effectson the human health on the long run [11] These conditionsoriented the search for natural bioactive compounds thathave the capabilities to control food borne pathogens
Horticultural crops tend to produce secondary metabo-lites during stress conditions such as water stressWater stressis one of the major limiting factors for agricultural industryespecially in view of the rapidly increasing world populationglobal climate change and the increasing worldwide indus-trial demand for water [12] Water stress may have severalmorphological (eg leaf number and leaf area) physiological(eg carbohydrate and ion composition) metabolic (egSOD activity and composition) and molecular (eg freeradical scavenging gene products) effects on plants leadingto reduced yields as well as increased accumulation of severalcompounds Plant metabolic responses to water stress mayinclude the accumulation of carbohydrates [13] increasedsynthesis of specific proteins increased stress related nutrientuptake (eg K) and accumulation of specific antioxidantssuch as the phenolic compounds and others that neutralizereactive oxygen species (ROS) [14ndash17]
Efforts to develop novel tools to enable horticultural cropsto cope with water stress on plants are a growing concernworldwide such as the use of biochar [18] 120573-aminobutyricacid [19] trinexapac-ethyl [20] seaweed extracts [21]nanoparticles [22] and oligosaccharide Oligosaccharide isa biostimulant produced commercially by subjecting chitinto high temperature followed by deacetylation using alka-line conditions to remove proteins and calcium [23 24]Oligosaccharides may be formulated as a solution or aswater-soluble powder They are widely used as plant elicitorsof the production of secondary metabolites [25] particu-larly polyphenols [26] Oligosaccharides have also strongantimicrobial activities and may stimulate the growth ofbeneficiary microbes [27] Additionally several studies sug-gest that it may improve crop yield [23] and enhancestress tolerance [28] However little is known regarding themechanism whereby oligosaccharides enhance water stresstolerance and effect secondary metabolites in horticulturalcrops
Saxifragaceae includes 30 genera of herbaceous peren-nials such as Heuchera which are known to be geneticallydiverse due to hybridization [29] Heuchera contains about50 species One of these is Heuchera which accommodatesperennial herbaceous ornamental shade plants widely usedin North America Europe North Africa and South Asia[30] Dozens of colored hybrid cultivars varying in leaf andflower color have been recently introduced in the marketInterestingly although native people of Europe have usedHeuchera and other genera of Saxifragaceae as traditionalmedicinal plants [31] for centuries the medicinal propertiesresponses of this species to oligosaccharide elicitors underwater stress have not been investigated
In the present study our objective was to explore thepossible effects of oligosaccharides onHeuchera grown undernormal and prolonged irrigation intervals by using morpho-logical physiological and metabolic markers We hypothe-sized that stress conditions and oligosaccharides treatmentmay enhance antimicrobial properties of Heuchera plantsThe information obtained from this study will contribute toour understanding of oligosaccharides andor water stressaction in plant metabolic responses that may help in thediscovery and use of natural bioactive compounds controlfood spoilage microorganisms
2 Material and Methods
21 Plant Material and Treatments Young plants 10 cmhigh of Heuchera cultivars Creme Brulee and Mahoganywere obtained from local commercial nurseries on January7th 2017 and 2018 Plants were grown in a polyethylene-covered greenhouse located on the Alexandria-Cairo desertroad Egypt All plants were identified by Hosam Elansaryand registered at the Faculty of Agriculture AlexandriaUniversity prior to transplanting onto 21 L pots containinga mixture of brown peat and perlite (31 ww) supplementedwith Crystalon (20 N 20 P 20 K 2 gL media) Plantswere grown for three weeks under temperatures rangingbetween 151∘C and 275∘C relative humidity between 58and 67 photosynthetically active radiation around 1000mminus2 at 1200 pm and daily watering of 38-50mLplant Plantswere divided into two groups one of which was watered at2-day intervals (2DWI) while the other was watered at 6-dayintervals (6DWI) for 6weeks Oligosaccharide (deacetylationgt 95 MW501486 gmol powder Aldebeiky Group CoCairo Egypt) water solution was sprayed at concentrations of50 200 or 500 ppm until drop off 2 weeks prior to extendingthe watering interval untreated plants were considered asthe control treatment The experiment was laid out in asplit-plot design Irrigation intervals were considered as themain plot and oligosaccharides treatments the subplot Plantswere grouped into three blocksrepetitions (n=3) containing5 replicates per treatment for a total of 40 plants per cultivarper season in Randomized Complete Block Design (RCBD)
22 Morphological and Physiological Parameters Plants wereharvested after 6 weeks of stress treatment At that pointplant height and leaf number were registered Leaf area wascalculated immediately using a scanner and the AutoCADprogram Total dry weight was determined by drying cleanedplants to constant weight in an oven at 70∘C Total carbohy-drates K+ Ca2+ and proline were determined in plant leavesat the end of the experiment Following freeze-drying ofsamples they were ground and sieved and then kept at -20∘Cuntil further analysis Total carbohydrates were quantifiedafter Dubios et al [32] and expressed on a percent basis Onegram of frozen leaves was used to obtain the cell sap thena dilution (1100 vv) was used for the determination of K+andCa2+ concentrations using an inductively coupled plasmaspectrophotometer [33] Proline leaf content was determinedin the Department of Plant Production King Saud Universityusing a spectrophotometer at 520 nm [34 35]
Evidence-Based Complementary and Alternative Medicine 3
23 Antioxidants Chlorophyll Phenols andEnzymeActivitiesAir dried leaves were ground into fine powder 025 g of thisfrom each sample was dissolved into 3 mL methanol (99)while stirring on a magnetic agitator at low speed in thedark for 24 h at room temperature Methanolic extracts werecentrifuged for 5 min under cooling at 10000 RPM (7000 timesg) the supernatant (sim27mL)was dried in a rotary evaporatorto produce a semisolid extract which was stored for laterantioxidant analysis Antioxidant activities of all sampleswere determined in the Department of Plant ProducitonKing Saud University using the 221015840-diphenylpicrylhydrazyl(DPPH) and 120573-carotene-linoleic acid methods which mea-sure OHminus scavenging activities according to Elansary et al[21] For the DPPH method samples were incubated for 30min after which absorbance was measured at 517 nm Forthe 120573-carotene-linoleic acid assay absorbance was measuredat 470 nm The sample concentration required to scavenge50 of DPPH 120573-carotene-linoleic acid (IC
50in 120583gmL)
was determined by plotting the inhibition percentage againstextract concentration Butylated hydroxytoluene (BHT) wasused as a positive control and experiments were repeatedtwice in triplicate Total phenolic content in methanolicleaf extracts were performed using the Folin-Ciocalteaucolorimetric method using gallic acid as the reference andexpressing the results as gallic acid equivalents (mg GAEgminus1 ext) [36 37] Total chlorophyll content was quantified infresh leaves according to Moran and Porath [38]
Ground-frozen leaves were used to quantify total andfree ascorbate after Elansary et al [21] Briefly 05 g ofground-frozen leaf tissues were homogenized in 8 mL cooledtrichloroacetic acid (TCA 5 wv) next the mixture wascentrifuged for 10 min (10000 times g) at 4∘C The supernatantwas incubated with a mixture of PBS (200 mM pH 74)and dithiothreitol (DTT 15 mM) for 50 min excess DTTwas removed by adding N-ethylmaleimide (NEM 200 120583L05 wv) The solution was then mixed with TCA (1 mL10 wv) o-phosphoric acid (800 120583L 42 wv) and 22-dipyridyl in 70 (vv) ethanol (800 120583L 65 mM) and iron(III)chloride (400 120583L 3 wv) and incubated for 1 h at 42∘CAbsorbance by the mixture was measured at 525 nm Freeascorbate was determined using the same procedure exceptDTT and NEM were replaced with 400 120583L deionized waterwhile free and total ascorbate contents were determined usingstandard curves
Catalase (CAT) ascorbate peroxidase (APX) and super-oxide dismutase (SOD) activities as well as H
2O2accumula-
tion were quantified in leaves tissues following Elansary et al[21]
24Microorganisms andMedicinal Properties Themedicinalproperties of methanolic leaf extracts were studied againstselected pathogenic bacteria and fungi The selected bacteriawere Listeria monocytogenes (clinical isolate) Bacillus cereus(ATCC 14579) Staphylococcus aureus (ATCC 6538) Micro-coccus flavus (ATCC 10240) Pseudomonas aeruginosa (ATCC27853) and Escherichia coli (ATCC 35210)The selected fungiwere Aspergillus niger (ATCC 6275) A ochraceus (ATCC12066) A flavus (ATCC 9643) Penicillium ochrochloron(ATCC 48663) and Candida albicans (ATCC 12066) The
microdilution method [39] was used to determine theantibacterial and antifungal activities In the antibacterialassay the minimum inhibitory bactericidal concentration(MIC) was defined as the lowest concentration resultingin growth stop of the bacteria at the binocular level Theminimum bactericidal concentration (MBC) was definedas the lowest concentration resulting in killing 995 ofthe original inoculum Also the MBC was determined byserial subcultivation of the bacterial using 01-02 mgmL ofbacterial solution added to 100 120583L of TSB and incubatedfor one day In the antifungal activity assay the minimuminhibitory concentration (MIC) was defined as the lowestconcentration inhibiting the fungal growth at the binocularlevel while the minimum fungicidal concentration (MFC)was determined using subcultivations of the fungi (01-40mgmL) and was defined as the concentration killing 995of the original inoculum Experiments were performed twiceand negative controls (5 DMSO) as well as positive con-trols [antibacterial assay streptomycin and ampicillin 001-10 mgmL antifungal Fluconazole (FLZ) and ketoconazole(KLZ)] were used Experiments were repeated twice
25 Statistical Analyses The data obtained during the twogrowing seasons in 2017 and 2018 were expressed as meansand Least Significant Difference (LSD) was determined usingthe one way ANOVA test in SPSS (PASWVer 21) at P le 005
3 Results
31 Morphological and Physiological Responses to IrrigationIntervals and Oligosaccharide Increasing watering intervalsfrom 2 to 6 days significantly reduced morphological param-eters in bothHeuchera cultivars tested including leaf numberleaf area plant dry weight and plant height (Table 1) Inter-estingly under the normal irrigation interval (2DWI) theapplication of the oligosaccharide at 50 and 200 ppm signifi-cantly increased leaf number and area plant dry weight andplant height in both cultivars treated plants in both seasonscompared to untreated plants Further under prolongedirrigation interval (6DWI) there were significant increasesin both Creme Brulee and Mahogany in all morphologicalparameters measure in plants treated with oligosaccharide at50 and 200 ppm compared to oligosaccharide at 500 ppmand control treatment Prolonged irrigation interval (6DWI)significantly reduced total carbohydrates K Ca and prolinecontents in plants of both Creme Brulee and Mahoganycompared to the normal irrigation interval (2DWI) as shownin Table 2 Under 2DWI as well as 6DWI total carbohydratesK Ca and proline contents increased significantly in theleaves of oligosaccharides -treated plants at 50 and 200ppm compared to controls and 500 ppm oligosaccharidetreatment in both growing seasons
32 General Antioxidants Phenolics and ChlorophyllsExtension of irrigation interval from 2 to 6 days caused asignificant increase in DPPH free radical scavenging activityin both Heuchera cultivars (Table 3) The DPPH (IC
50) of
Creme Brulee plants decreased in the first season (2017)which indicates an increase in scavenging activity a similar
4 Evidence-Based Complementary and Alternative Medicine
Table1Eff
ecto
fwater
deficitandoligosaccharides
treatmento
nleafnu
mberleafareaplant
drywe
ightand
planth
eigh
tintwoHeucheracultivarsaft
ersix
weekso
ftreatmentinitia
tion
Values
aree
xpressed
asmeans
(plusmnsd)
Waterinterval
Oligosaccharides
treatment(pp
m)
Leafnu
mber(leafplantminus1)
Leafarea
(cm2plantminus1)
Plantd
rywe
ight
(gplantminus1)
Planth
eigh
t(cm
)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
156plusmn02blowast
152plusmn01b
6512plusmn151b
6482plusmn111b
112plusmn01b
112plusmn02b
291plusmn01b
288plusmn02b
50161plusmn01ab
161plusmn02a
6901plusmn
131a
6863plusmn145a
123plusmn01a
121plusmn02a
332plusmn01a
308plusmn03a
200
171plusmn
04a
172plusmn01a
7031plusmn
143a
6992plusmn151a
124plusmn01a
122plusmn02a
324plusmn02a
307plusmn02a
500
156plusmn01b
153plusmn02b
6392plusmn111b
6505plusmn
223b
113plusmn01b
112plusmn02b
301plusmn01b
294plusmn01b
6DWI
070plusmn01d
71plusmn02d
3031plusmn
103d
3111plusmn131d
55plusmn01d
56plusmn01d
173plusmn
03d
168plusmn01d
5086plusmn00cd
85plusmn03cd
3509plusmn151c
3581plusmn
112c
63plusmn01c
62plusmn01c
194plusmn01c
188plusmn01c
200
90plusmn01c
91plusmn02c
3613plusmn141c
3516plusmn175c
61plusmn
02c
62plusmn01c
195plusmn01c
190plusmn01c
500
73plusmn00d
72plusmn01d
3112plusmn131d
3061plusmn
141d
53plusmn01d
53plusmn01d
174plusmn02cd
172plusmn01d
2DWI
0Mahogany
132plusmn01b
128plusmn01b
5161plusmn
221b
5125plusmn101b
108plusmn03b
107plusmn01b
308plusmn03b
311plusmn04b
50144plusmn01a
142plusmn02a
5631plusmn
201a
5675plusmn112a
116plusmn01a
117plusmn02a
333plusmn01a
329plusmn03a
200
147plusmn03a
142plusmn03a
5731plusmn
103a
5781plusmn
121a
116plusmn01a
117plusmn01a
342plusmn04a
332plusmn03a
500
132plusmn01b
131plusmn01b
5103plusmn113b
5021plusmn
167b
108plusmn01b
108plusmn02b
314plusmn01b
307plusmn01b
6DWI
061plusmn
02e
62plusmn02e
2201plusmn
121d
2153plusmn152d
54plusmn01d
55plusmn01d
181plusmn01d
183plusmn03d
5074plusmn01d
71plusmn01d
2611plusmn131c
2683plusmn
113c
63plusmn01c
62plusmn01c
212plusmn03c
207plusmn01c
200
81plusmn
01 c
82plusmn02c
2713plusmn101c
2773plusmn111c
63plusmn01c
63plusmn01c
205plusmn02c
209plusmn01c
500
62plusmn01e
61plusmn
01e
2102plusmn115d
2215plusmn151d
55plusmn01d
54plusmn01d
183plusmn01d
187plusmn03d
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 5
Table2Eff
ectofirrigationintervalsand
oligosaccharidestre
atmentontotalcarbo
hydrateK
Caandprolinec
ontent
intheleaveso
ftwoHeucheracultivarsin
twosuccessiv
eseasonsV
alues
arem
eans
(plusmnsd)
Water
interval
Oligosaccharides
treatment
(ppm
)
Totalcarbo
hydrates
(DW)
K(m
ggminus1DW)
Ca(
mggminus1DW)
Proline(
mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
1345plusmn01blowast
1337plusmn01b
197plusmn01d
195plusmn05d
376plusmn005b
363plusmn004
b13
5plusmn005c
132plusmn001c
501433plusmn01a
1422plusmn01ab
248plusmn01b
239plusmn01b
412plusmn00a
411plusmn02a
144plusmn003b
141plusmn
000
cb200
1453plusmn02a
1467plusmn01a
258plusmn02b
249plusmn01b
415plusmn009a
409plusmn003a
147plusmn001b
144plusmn003b
500
1353plusmn01b
1349plusmn01b
199plusmn00d
194plusmn01d
385plusmn001b
379plusmn005b
137plusmn001c
135plusmn002c
6DWI
01219plusmn02c
1205plusmn01c
215plusmn01b
213plusmn03c
363plusmn006
b365plusmn006
b14
8plusmn002b
146plusmn001ab
501289plusmn01cb
1251plusmn
01c
273plusmn03a
268plusmn01a
405plusmn003a
397plusmn001a
156plusmn003a
153plusmn003a
200
1287plusmn01cb
1298plusmn01bc
279plusmn01a
277plusmn01a
419plusmn007a
411plusmn004
a15
8plusmn002a
155plusmn
002a
500
1232plusmn02c
1231plusmn01c
222plusmn02c
218plusmn02c
378plusmn006
b370plusmn005b
149plusmn003ab
147plusmn001ab
2DWI
0Mahogany
1532plusmn01b
a1505plusmn01b
213plusmn01c
217plusmn03c
361plusmn004
b367plusmn008b
142plusmn002c
138plusmn001c
501599plusmn01ab
1591plusmn
00a
264plusmn01b
261plusmn01b
397plusmn003a
394plusmn005a
155plusmn004
b14
6plusmn001b
200
1630plusmn01a
1613plusmn03a
268plusmn01b
266plusmn03b
406plusmn006
a401plusmn006
a15
8plusmn008b
149plusmn004
b500
1551plusmn
01b
1527plusmn03b
233plusmn03d
226plusmn01c
373plusmn004
b375plusmn003b
147plusmn001c
141plusmn
001c
6DWI
01441plusmn
02c
1412plusmn03c
255plusmn01b
247plusmn01b
358plusmn002b
368plusmn002b
156plusmn003b
151plusmn
004
b50
1545plusmn01b
1513plusmn01b
299plusmn01a
289plusmn03a
393plusmn001a
395plusmn001a
170plusmn001a
169plusmn002a
200
1555plusmn01b
1525plusmn05b
3 03plusmn01a
297plusmn02a
393plusmn001a
396plusmn004
a174plusmn003a
171plusmn
002a
500
1437plusmn01c
1424plusmn03c
264plusmn01a
258plusmn03b
363plusmn005b
367plusmn002b
159plusmn002b
154plusmn001b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
6 Evidence-Based Complementary and Alternative Medicine
Table3Antioxidant
activ
ityin
leafmethano
licextracts
totalpheno
licandtotalchlorop
hyllcontento
ftwo
HeucheracultivarsV
aluesa
remeans
oftriplicated
eterminationsplusmnsd
Water
interval
Oligosaccharides
treatment
(ppm
)
DPP
Hfre
eradical
scavenging
activ
ity(IC
50120583
gmlminus1
)
120573-C
arotene-lin
oleica
cid
assay
(IC50120583
gmlminus1
)
Totalp
heno
liccontent
(mgGAEgminus1)
Totalchlorop
hyllcontent
(mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
103plusmn001alowast
111plusmn
007a
112plusmn001a
115plusmn001a
104plusmn01c
97plusmn01c
065plusmn004
b063plusmn001bc
5093plusmn006
b99plusmn001b
103plusmn003b
104plusmn002b
109plusmn01b
104plusmn03b
069plusmn002a
067plusmn003a
200
91plusmn005b
99plusmn002b
103plusmn003b
107plusmn003b
108plusmn00b
105plusmn02b
070plusmn002a
068plusmn001a
500
96plusmn003a
104plusmn003a
112plusmn002a
115plusmn004
a104plusmn02c
100plusmn01c
068plusmn001ab
065plusmn002ab
6DWI
082plusmn004
c89plusmn005c
93plusmn001c
101plusmn003c
109plusmn04b
104plusmn02b
061plusmn002c
060plusmn003c
5063plusmn002d
68plusmn000
d74plusmn002d
82plusmn002d
116plusmn03a
112plusmn02a
065plusmn001b
064plusmn002b
200
62plusmn001d
68plusmn001d
72plusmn003d
82plusmn002d
116plusmn01a
113plusmn02a
065plusmn001b
065plusmn003ab
500
78plusmn003c
83plusmn001c
92plusmn004
c99plusmn001c
111plusmn
02b
105plusmn04b
062plusmn002c
061plusmn001c
2DWI
0Mahogany
89plusmn04a
96plusmn01a
101plusmn03a
107plusmn02a
123plusmn02c
117plusmn03c
071plusmn001b
070plusmn002b
5074plusmn002b
79plusmn001b
85plusmn001b
94plusmn003b
127plusmn03b
124plusmn01b
075plusmn002a
074plusmn001a
200
71plusmn001b
78plusmn004
b84plusmn000
b93plusmn003b
129plusmn04b
125plusmn02b
076plusmn001a
075plusmn001a
500
82plusmn002a
88plusmn003a
95plusmn003a
104plusmn002a
122plusmn01c
121plusmn03b
c072plusmn001b
070plusmn002b
6DWI
071plusmn003b
77plusmn002b
84plusmn001b
89plusmn003b
131plusmn01b
124plusmn02b
066plusmn002c
063plusmn001c
5054plusmn001c
57plusmn002c
66plusmn003c
72plusmn003c
138plusmn02a
132plusmn00a
070plusmn001c
068plusmn003c
200
48plusmn00 3c
55plusmn007c
62plusmn002c
71plusmn002c
139plusmn03a
134plusmn01a
071plusmn001a
068plusmn002a
500
68plusmn001b
74plusmn003b
79plusmn003b
82plusmn001b
134plusmn02ab
125plusmn01b
068plusmn002b
064plusmn003b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
different
basedon
LSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
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Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 3
23 Antioxidants Chlorophyll Phenols andEnzymeActivitiesAir dried leaves were ground into fine powder 025 g of thisfrom each sample was dissolved into 3 mL methanol (99)while stirring on a magnetic agitator at low speed in thedark for 24 h at room temperature Methanolic extracts werecentrifuged for 5 min under cooling at 10000 RPM (7000 timesg) the supernatant (sim27mL)was dried in a rotary evaporatorto produce a semisolid extract which was stored for laterantioxidant analysis Antioxidant activities of all sampleswere determined in the Department of Plant ProducitonKing Saud University using the 221015840-diphenylpicrylhydrazyl(DPPH) and 120573-carotene-linoleic acid methods which mea-sure OHminus scavenging activities according to Elansary et al[21] For the DPPH method samples were incubated for 30min after which absorbance was measured at 517 nm Forthe 120573-carotene-linoleic acid assay absorbance was measuredat 470 nm The sample concentration required to scavenge50 of DPPH 120573-carotene-linoleic acid (IC
50in 120583gmL)
was determined by plotting the inhibition percentage againstextract concentration Butylated hydroxytoluene (BHT) wasused as a positive control and experiments were repeatedtwice in triplicate Total phenolic content in methanolicleaf extracts were performed using the Folin-Ciocalteaucolorimetric method using gallic acid as the reference andexpressing the results as gallic acid equivalents (mg GAEgminus1 ext) [36 37] Total chlorophyll content was quantified infresh leaves according to Moran and Porath [38]
Ground-frozen leaves were used to quantify total andfree ascorbate after Elansary et al [21] Briefly 05 g ofground-frozen leaf tissues were homogenized in 8 mL cooledtrichloroacetic acid (TCA 5 wv) next the mixture wascentrifuged for 10 min (10000 times g) at 4∘C The supernatantwas incubated with a mixture of PBS (200 mM pH 74)and dithiothreitol (DTT 15 mM) for 50 min excess DTTwas removed by adding N-ethylmaleimide (NEM 200 120583L05 wv) The solution was then mixed with TCA (1 mL10 wv) o-phosphoric acid (800 120583L 42 wv) and 22-dipyridyl in 70 (vv) ethanol (800 120583L 65 mM) and iron(III)chloride (400 120583L 3 wv) and incubated for 1 h at 42∘CAbsorbance by the mixture was measured at 525 nm Freeascorbate was determined using the same procedure exceptDTT and NEM were replaced with 400 120583L deionized waterwhile free and total ascorbate contents were determined usingstandard curves
Catalase (CAT) ascorbate peroxidase (APX) and super-oxide dismutase (SOD) activities as well as H
2O2accumula-
tion were quantified in leaves tissues following Elansary et al[21]
24Microorganisms andMedicinal Properties Themedicinalproperties of methanolic leaf extracts were studied againstselected pathogenic bacteria and fungi The selected bacteriawere Listeria monocytogenes (clinical isolate) Bacillus cereus(ATCC 14579) Staphylococcus aureus (ATCC 6538) Micro-coccus flavus (ATCC 10240) Pseudomonas aeruginosa (ATCC27853) and Escherichia coli (ATCC 35210)The selected fungiwere Aspergillus niger (ATCC 6275) A ochraceus (ATCC12066) A flavus (ATCC 9643) Penicillium ochrochloron(ATCC 48663) and Candida albicans (ATCC 12066) The
microdilution method [39] was used to determine theantibacterial and antifungal activities In the antibacterialassay the minimum inhibitory bactericidal concentration(MIC) was defined as the lowest concentration resultingin growth stop of the bacteria at the binocular level Theminimum bactericidal concentration (MBC) was definedas the lowest concentration resulting in killing 995 ofthe original inoculum Also the MBC was determined byserial subcultivation of the bacterial using 01-02 mgmL ofbacterial solution added to 100 120583L of TSB and incubatedfor one day In the antifungal activity assay the minimuminhibitory concentration (MIC) was defined as the lowestconcentration inhibiting the fungal growth at the binocularlevel while the minimum fungicidal concentration (MFC)was determined using subcultivations of the fungi (01-40mgmL) and was defined as the concentration killing 995of the original inoculum Experiments were performed twiceand negative controls (5 DMSO) as well as positive con-trols [antibacterial assay streptomycin and ampicillin 001-10 mgmL antifungal Fluconazole (FLZ) and ketoconazole(KLZ)] were used Experiments were repeated twice
25 Statistical Analyses The data obtained during the twogrowing seasons in 2017 and 2018 were expressed as meansand Least Significant Difference (LSD) was determined usingthe one way ANOVA test in SPSS (PASWVer 21) at P le 005
3 Results
31 Morphological and Physiological Responses to IrrigationIntervals and Oligosaccharide Increasing watering intervalsfrom 2 to 6 days significantly reduced morphological param-eters in bothHeuchera cultivars tested including leaf numberleaf area plant dry weight and plant height (Table 1) Inter-estingly under the normal irrigation interval (2DWI) theapplication of the oligosaccharide at 50 and 200 ppm signifi-cantly increased leaf number and area plant dry weight andplant height in both cultivars treated plants in both seasonscompared to untreated plants Further under prolongedirrigation interval (6DWI) there were significant increasesin both Creme Brulee and Mahogany in all morphologicalparameters measure in plants treated with oligosaccharide at50 and 200 ppm compared to oligosaccharide at 500 ppmand control treatment Prolonged irrigation interval (6DWI)significantly reduced total carbohydrates K Ca and prolinecontents in plants of both Creme Brulee and Mahoganycompared to the normal irrigation interval (2DWI) as shownin Table 2 Under 2DWI as well as 6DWI total carbohydratesK Ca and proline contents increased significantly in theleaves of oligosaccharides -treated plants at 50 and 200ppm compared to controls and 500 ppm oligosaccharidetreatment in both growing seasons
32 General Antioxidants Phenolics and ChlorophyllsExtension of irrigation interval from 2 to 6 days caused asignificant increase in DPPH free radical scavenging activityin both Heuchera cultivars (Table 3) The DPPH (IC
50) of
Creme Brulee plants decreased in the first season (2017)which indicates an increase in scavenging activity a similar
4 Evidence-Based Complementary and Alternative Medicine
Table1Eff
ecto
fwater
deficitandoligosaccharides
treatmento
nleafnu
mberleafareaplant
drywe
ightand
planth
eigh
tintwoHeucheracultivarsaft
ersix
weekso
ftreatmentinitia
tion
Values
aree
xpressed
asmeans
(plusmnsd)
Waterinterval
Oligosaccharides
treatment(pp
m)
Leafnu
mber(leafplantminus1)
Leafarea
(cm2plantminus1)
Plantd
rywe
ight
(gplantminus1)
Planth
eigh
t(cm
)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
156plusmn02blowast
152plusmn01b
6512plusmn151b
6482plusmn111b
112plusmn01b
112plusmn02b
291plusmn01b
288plusmn02b
50161plusmn01ab
161plusmn02a
6901plusmn
131a
6863plusmn145a
123plusmn01a
121plusmn02a
332plusmn01a
308plusmn03a
200
171plusmn
04a
172plusmn01a
7031plusmn
143a
6992plusmn151a
124plusmn01a
122plusmn02a
324plusmn02a
307plusmn02a
500
156plusmn01b
153plusmn02b
6392plusmn111b
6505plusmn
223b
113plusmn01b
112plusmn02b
301plusmn01b
294plusmn01b
6DWI
070plusmn01d
71plusmn02d
3031plusmn
103d
3111plusmn131d
55plusmn01d
56plusmn01d
173plusmn
03d
168plusmn01d
5086plusmn00cd
85plusmn03cd
3509plusmn151c
3581plusmn
112c
63plusmn01c
62plusmn01c
194plusmn01c
188plusmn01c
200
90plusmn01c
91plusmn02c
3613plusmn141c
3516plusmn175c
61plusmn
02c
62plusmn01c
195plusmn01c
190plusmn01c
500
73plusmn00d
72plusmn01d
3112plusmn131d
3061plusmn
141d
53plusmn01d
53plusmn01d
174plusmn02cd
172plusmn01d
2DWI
0Mahogany
132plusmn01b
128plusmn01b
5161plusmn
221b
5125plusmn101b
108plusmn03b
107plusmn01b
308plusmn03b
311plusmn04b
50144plusmn01a
142plusmn02a
5631plusmn
201a
5675plusmn112a
116plusmn01a
117plusmn02a
333plusmn01a
329plusmn03a
200
147plusmn03a
142plusmn03a
5731plusmn
103a
5781plusmn
121a
116plusmn01a
117plusmn01a
342plusmn04a
332plusmn03a
500
132plusmn01b
131plusmn01b
5103plusmn113b
5021plusmn
167b
108plusmn01b
108plusmn02b
314plusmn01b
307plusmn01b
6DWI
061plusmn
02e
62plusmn02e
2201plusmn
121d
2153plusmn152d
54plusmn01d
55plusmn01d
181plusmn01d
183plusmn03d
5074plusmn01d
71plusmn01d
2611plusmn131c
2683plusmn
113c
63plusmn01c
62plusmn01c
212plusmn03c
207plusmn01c
200
81plusmn
01 c
82plusmn02c
2713plusmn101c
2773plusmn111c
63plusmn01c
63plusmn01c
205plusmn02c
209plusmn01c
500
62plusmn01e
61plusmn
01e
2102plusmn115d
2215plusmn151d
55plusmn01d
54plusmn01d
183plusmn01d
187plusmn03d
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 5
Table2Eff
ectofirrigationintervalsand
oligosaccharidestre
atmentontotalcarbo
hydrateK
Caandprolinec
ontent
intheleaveso
ftwoHeucheracultivarsin
twosuccessiv
eseasonsV
alues
arem
eans
(plusmnsd)
Water
interval
Oligosaccharides
treatment
(ppm
)
Totalcarbo
hydrates
(DW)
K(m
ggminus1DW)
Ca(
mggminus1DW)
Proline(
mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
1345plusmn01blowast
1337plusmn01b
197plusmn01d
195plusmn05d
376plusmn005b
363plusmn004
b13
5plusmn005c
132plusmn001c
501433plusmn01a
1422plusmn01ab
248plusmn01b
239plusmn01b
412plusmn00a
411plusmn02a
144plusmn003b
141plusmn
000
cb200
1453plusmn02a
1467plusmn01a
258plusmn02b
249plusmn01b
415plusmn009a
409plusmn003a
147plusmn001b
144plusmn003b
500
1353plusmn01b
1349plusmn01b
199plusmn00d
194plusmn01d
385plusmn001b
379plusmn005b
137plusmn001c
135plusmn002c
6DWI
01219plusmn02c
1205plusmn01c
215plusmn01b
213plusmn03c
363plusmn006
b365plusmn006
b14
8plusmn002b
146plusmn001ab
501289plusmn01cb
1251plusmn
01c
273plusmn03a
268plusmn01a
405plusmn003a
397plusmn001a
156plusmn003a
153plusmn003a
200
1287plusmn01cb
1298plusmn01bc
279plusmn01a
277plusmn01a
419plusmn007a
411plusmn004
a15
8plusmn002a
155plusmn
002a
500
1232plusmn02c
1231plusmn01c
222plusmn02c
218plusmn02c
378plusmn006
b370plusmn005b
149plusmn003ab
147plusmn001ab
2DWI
0Mahogany
1532plusmn01b
a1505plusmn01b
213plusmn01c
217plusmn03c
361plusmn004
b367plusmn008b
142plusmn002c
138plusmn001c
501599plusmn01ab
1591plusmn
00a
264plusmn01b
261plusmn01b
397plusmn003a
394plusmn005a
155plusmn004
b14
6plusmn001b
200
1630plusmn01a
1613plusmn03a
268plusmn01b
266plusmn03b
406plusmn006
a401plusmn006
a15
8plusmn008b
149plusmn004
b500
1551plusmn
01b
1527plusmn03b
233plusmn03d
226plusmn01c
373plusmn004
b375plusmn003b
147plusmn001c
141plusmn
001c
6DWI
01441plusmn
02c
1412plusmn03c
255plusmn01b
247plusmn01b
358plusmn002b
368plusmn002b
156plusmn003b
151plusmn
004
b50
1545plusmn01b
1513plusmn01b
299plusmn01a
289plusmn03a
393plusmn001a
395plusmn001a
170plusmn001a
169plusmn002a
200
1555plusmn01b
1525plusmn05b
3 03plusmn01a
297plusmn02a
393plusmn001a
396plusmn004
a174plusmn003a
171plusmn
002a
500
1437plusmn01c
1424plusmn03c
264plusmn01a
258plusmn03b
363plusmn005b
367plusmn002b
159plusmn002b
154plusmn001b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
6 Evidence-Based Complementary and Alternative Medicine
Table3Antioxidant
activ
ityin
leafmethano
licextracts
totalpheno
licandtotalchlorop
hyllcontento
ftwo
HeucheracultivarsV
aluesa
remeans
oftriplicated
eterminationsplusmnsd
Water
interval
Oligosaccharides
treatment
(ppm
)
DPP
Hfre
eradical
scavenging
activ
ity(IC
50120583
gmlminus1
)
120573-C
arotene-lin
oleica
cid
assay
(IC50120583
gmlminus1
)
Totalp
heno
liccontent
(mgGAEgminus1)
Totalchlorop
hyllcontent
(mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
103plusmn001alowast
111plusmn
007a
112plusmn001a
115plusmn001a
104plusmn01c
97plusmn01c
065plusmn004
b063plusmn001bc
5093plusmn006
b99plusmn001b
103plusmn003b
104plusmn002b
109plusmn01b
104plusmn03b
069plusmn002a
067plusmn003a
200
91plusmn005b
99plusmn002b
103plusmn003b
107plusmn003b
108plusmn00b
105plusmn02b
070plusmn002a
068plusmn001a
500
96plusmn003a
104plusmn003a
112plusmn002a
115plusmn004
a104plusmn02c
100plusmn01c
068plusmn001ab
065plusmn002ab
6DWI
082plusmn004
c89plusmn005c
93plusmn001c
101plusmn003c
109plusmn04b
104plusmn02b
061plusmn002c
060plusmn003c
5063plusmn002d
68plusmn000
d74plusmn002d
82plusmn002d
116plusmn03a
112plusmn02a
065plusmn001b
064plusmn002b
200
62plusmn001d
68plusmn001d
72plusmn003d
82plusmn002d
116plusmn01a
113plusmn02a
065plusmn001b
065plusmn003ab
500
78plusmn003c
83plusmn001c
92plusmn004
c99plusmn001c
111plusmn
02b
105plusmn04b
062plusmn002c
061plusmn001c
2DWI
0Mahogany
89plusmn04a
96plusmn01a
101plusmn03a
107plusmn02a
123plusmn02c
117plusmn03c
071plusmn001b
070plusmn002b
5074plusmn002b
79plusmn001b
85plusmn001b
94plusmn003b
127plusmn03b
124plusmn01b
075plusmn002a
074plusmn001a
200
71plusmn001b
78plusmn004
b84plusmn000
b93plusmn003b
129plusmn04b
125plusmn02b
076plusmn001a
075plusmn001a
500
82plusmn002a
88plusmn003a
95plusmn003a
104plusmn002a
122plusmn01c
121plusmn03b
c072plusmn001b
070plusmn002b
6DWI
071plusmn003b
77plusmn002b
84plusmn001b
89plusmn003b
131plusmn01b
124plusmn02b
066plusmn002c
063plusmn001c
5054plusmn001c
57plusmn002c
66plusmn003c
72plusmn003c
138plusmn02a
132plusmn00a
070plusmn001c
068plusmn003c
200
48plusmn00 3c
55plusmn007c
62plusmn002c
71plusmn002c
139plusmn03a
134plusmn01a
071plusmn001a
068plusmn002a
500
68plusmn001b
74plusmn003b
79plusmn003b
82plusmn001b
134plusmn02ab
125plusmn01b
068plusmn002b
064plusmn003b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
different
basedon
LSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
4 Evidence-Based Complementary and Alternative Medicine
Table1Eff
ecto
fwater
deficitandoligosaccharides
treatmento
nleafnu
mberleafareaplant
drywe
ightand
planth
eigh
tintwoHeucheracultivarsaft
ersix
weekso
ftreatmentinitia
tion
Values
aree
xpressed
asmeans
(plusmnsd)
Waterinterval
Oligosaccharides
treatment(pp
m)
Leafnu
mber(leafplantminus1)
Leafarea
(cm2plantminus1)
Plantd
rywe
ight
(gplantminus1)
Planth
eigh
t(cm
)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
156plusmn02blowast
152plusmn01b
6512plusmn151b
6482plusmn111b
112plusmn01b
112plusmn02b
291plusmn01b
288plusmn02b
50161plusmn01ab
161plusmn02a
6901plusmn
131a
6863plusmn145a
123plusmn01a
121plusmn02a
332plusmn01a
308plusmn03a
200
171plusmn
04a
172plusmn01a
7031plusmn
143a
6992plusmn151a
124plusmn01a
122plusmn02a
324plusmn02a
307plusmn02a
500
156plusmn01b
153plusmn02b
6392plusmn111b
6505plusmn
223b
113plusmn01b
112plusmn02b
301plusmn01b
294plusmn01b
6DWI
070plusmn01d
71plusmn02d
3031plusmn
103d
3111plusmn131d
55plusmn01d
56plusmn01d
173plusmn
03d
168plusmn01d
5086plusmn00cd
85plusmn03cd
3509plusmn151c
3581plusmn
112c
63plusmn01c
62plusmn01c
194plusmn01c
188plusmn01c
200
90plusmn01c
91plusmn02c
3613plusmn141c
3516plusmn175c
61plusmn
02c
62plusmn01c
195plusmn01c
190plusmn01c
500
73plusmn00d
72plusmn01d
3112plusmn131d
3061plusmn
141d
53plusmn01d
53plusmn01d
174plusmn02cd
172plusmn01d
2DWI
0Mahogany
132plusmn01b
128plusmn01b
5161plusmn
221b
5125plusmn101b
108plusmn03b
107plusmn01b
308plusmn03b
311plusmn04b
50144plusmn01a
142plusmn02a
5631plusmn
201a
5675plusmn112a
116plusmn01a
117plusmn02a
333plusmn01a
329plusmn03a
200
147plusmn03a
142plusmn03a
5731plusmn
103a
5781plusmn
121a
116plusmn01a
117plusmn01a
342plusmn04a
332plusmn03a
500
132plusmn01b
131plusmn01b
5103plusmn113b
5021plusmn
167b
108plusmn01b
108plusmn02b
314plusmn01b
307plusmn01b
6DWI
061plusmn
02e
62plusmn02e
2201plusmn
121d
2153plusmn152d
54plusmn01d
55plusmn01d
181plusmn01d
183plusmn03d
5074plusmn01d
71plusmn01d
2611plusmn131c
2683plusmn
113c
63plusmn01c
62plusmn01c
212plusmn03c
207plusmn01c
200
81plusmn
01 c
82plusmn02c
2713plusmn101c
2773plusmn111c
63plusmn01c
63plusmn01c
205plusmn02c
209plusmn01c
500
62plusmn01e
61plusmn
01e
2102plusmn115d
2215plusmn151d
55plusmn01d
54plusmn01d
183plusmn01d
187plusmn03d
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 5
Table2Eff
ectofirrigationintervalsand
oligosaccharidestre
atmentontotalcarbo
hydrateK
Caandprolinec
ontent
intheleaveso
ftwoHeucheracultivarsin
twosuccessiv
eseasonsV
alues
arem
eans
(plusmnsd)
Water
interval
Oligosaccharides
treatment
(ppm
)
Totalcarbo
hydrates
(DW)
K(m
ggminus1DW)
Ca(
mggminus1DW)
Proline(
mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
1345plusmn01blowast
1337plusmn01b
197plusmn01d
195plusmn05d
376plusmn005b
363plusmn004
b13
5plusmn005c
132plusmn001c
501433plusmn01a
1422plusmn01ab
248plusmn01b
239plusmn01b
412plusmn00a
411plusmn02a
144plusmn003b
141plusmn
000
cb200
1453plusmn02a
1467plusmn01a
258plusmn02b
249plusmn01b
415plusmn009a
409plusmn003a
147plusmn001b
144plusmn003b
500
1353plusmn01b
1349plusmn01b
199plusmn00d
194plusmn01d
385plusmn001b
379plusmn005b
137plusmn001c
135plusmn002c
6DWI
01219plusmn02c
1205plusmn01c
215plusmn01b
213plusmn03c
363plusmn006
b365plusmn006
b14
8plusmn002b
146plusmn001ab
501289plusmn01cb
1251plusmn
01c
273plusmn03a
268plusmn01a
405plusmn003a
397plusmn001a
156plusmn003a
153plusmn003a
200
1287plusmn01cb
1298plusmn01bc
279plusmn01a
277plusmn01a
419plusmn007a
411plusmn004
a15
8plusmn002a
155plusmn
002a
500
1232plusmn02c
1231plusmn01c
222plusmn02c
218plusmn02c
378plusmn006
b370plusmn005b
149plusmn003ab
147plusmn001ab
2DWI
0Mahogany
1532plusmn01b
a1505plusmn01b
213plusmn01c
217plusmn03c
361plusmn004
b367plusmn008b
142plusmn002c
138plusmn001c
501599plusmn01ab
1591plusmn
00a
264plusmn01b
261plusmn01b
397plusmn003a
394plusmn005a
155plusmn004
b14
6plusmn001b
200
1630plusmn01a
1613plusmn03a
268plusmn01b
266plusmn03b
406plusmn006
a401plusmn006
a15
8plusmn008b
149plusmn004
b500
1551plusmn
01b
1527plusmn03b
233plusmn03d
226plusmn01c
373plusmn004
b375plusmn003b
147plusmn001c
141plusmn
001c
6DWI
01441plusmn
02c
1412plusmn03c
255plusmn01b
247plusmn01b
358plusmn002b
368plusmn002b
156plusmn003b
151plusmn
004
b50
1545plusmn01b
1513plusmn01b
299plusmn01a
289plusmn03a
393plusmn001a
395plusmn001a
170plusmn001a
169plusmn002a
200
1555plusmn01b
1525plusmn05b
3 03plusmn01a
297plusmn02a
393plusmn001a
396plusmn004
a174plusmn003a
171plusmn
002a
500
1437plusmn01c
1424plusmn03c
264plusmn01a
258plusmn03b
363plusmn005b
367plusmn002b
159plusmn002b
154plusmn001b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
6 Evidence-Based Complementary and Alternative Medicine
Table3Antioxidant
activ
ityin
leafmethano
licextracts
totalpheno
licandtotalchlorop
hyllcontento
ftwo
HeucheracultivarsV
aluesa
remeans
oftriplicated
eterminationsplusmnsd
Water
interval
Oligosaccharides
treatment
(ppm
)
DPP
Hfre
eradical
scavenging
activ
ity(IC
50120583
gmlminus1
)
120573-C
arotene-lin
oleica
cid
assay
(IC50120583
gmlminus1
)
Totalp
heno
liccontent
(mgGAEgminus1)
Totalchlorop
hyllcontent
(mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
103plusmn001alowast
111plusmn
007a
112plusmn001a
115plusmn001a
104plusmn01c
97plusmn01c
065plusmn004
b063plusmn001bc
5093plusmn006
b99plusmn001b
103plusmn003b
104plusmn002b
109plusmn01b
104plusmn03b
069plusmn002a
067plusmn003a
200
91plusmn005b
99plusmn002b
103plusmn003b
107plusmn003b
108plusmn00b
105plusmn02b
070plusmn002a
068plusmn001a
500
96plusmn003a
104plusmn003a
112plusmn002a
115plusmn004
a104plusmn02c
100plusmn01c
068plusmn001ab
065plusmn002ab
6DWI
082plusmn004
c89plusmn005c
93plusmn001c
101plusmn003c
109plusmn04b
104plusmn02b
061plusmn002c
060plusmn003c
5063plusmn002d
68plusmn000
d74plusmn002d
82plusmn002d
116plusmn03a
112plusmn02a
065plusmn001b
064plusmn002b
200
62plusmn001d
68plusmn001d
72plusmn003d
82plusmn002d
116plusmn01a
113plusmn02a
065plusmn001b
065plusmn003ab
500
78plusmn003c
83plusmn001c
92plusmn004
c99plusmn001c
111plusmn
02b
105plusmn04b
062plusmn002c
061plusmn001c
2DWI
0Mahogany
89plusmn04a
96plusmn01a
101plusmn03a
107plusmn02a
123plusmn02c
117plusmn03c
071plusmn001b
070plusmn002b
5074plusmn002b
79plusmn001b
85plusmn001b
94plusmn003b
127plusmn03b
124plusmn01b
075plusmn002a
074plusmn001a
200
71plusmn001b
78plusmn004
b84plusmn000
b93plusmn003b
129plusmn04b
125plusmn02b
076plusmn001a
075plusmn001a
500
82plusmn002a
88plusmn003a
95plusmn003a
104plusmn002a
122plusmn01c
121plusmn03b
c072plusmn001b
070plusmn002b
6DWI
071plusmn003b
77plusmn002b
84plusmn001b
89plusmn003b
131plusmn01b
124plusmn02b
066plusmn002c
063plusmn001c
5054plusmn001c
57plusmn002c
66plusmn003c
72plusmn003c
138plusmn02a
132plusmn00a
070plusmn001c
068plusmn003c
200
48plusmn00 3c
55plusmn007c
62plusmn002c
71plusmn002c
139plusmn03a
134plusmn01a
071plusmn001a
068plusmn002a
500
68plusmn001b
74plusmn003b
79plusmn003b
82plusmn001b
134plusmn02ab
125plusmn01b
068plusmn002b
064plusmn003b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
different
basedon
LSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 5
Table2Eff
ectofirrigationintervalsand
oligosaccharidestre
atmentontotalcarbo
hydrateK
Caandprolinec
ontent
intheleaveso
ftwoHeucheracultivarsin
twosuccessiv
eseasonsV
alues
arem
eans
(plusmnsd)
Water
interval
Oligosaccharides
treatment
(ppm
)
Totalcarbo
hydrates
(DW)
K(m
ggminus1DW)
Ca(
mggminus1DW)
Proline(
mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
1345plusmn01blowast
1337plusmn01b
197plusmn01d
195plusmn05d
376plusmn005b
363plusmn004
b13
5plusmn005c
132plusmn001c
501433plusmn01a
1422plusmn01ab
248plusmn01b
239plusmn01b
412plusmn00a
411plusmn02a
144plusmn003b
141plusmn
000
cb200
1453plusmn02a
1467plusmn01a
258plusmn02b
249plusmn01b
415plusmn009a
409plusmn003a
147plusmn001b
144plusmn003b
500
1353plusmn01b
1349plusmn01b
199plusmn00d
194plusmn01d
385plusmn001b
379plusmn005b
137plusmn001c
135plusmn002c
6DWI
01219plusmn02c
1205plusmn01c
215plusmn01b
213plusmn03c
363plusmn006
b365plusmn006
b14
8plusmn002b
146plusmn001ab
501289plusmn01cb
1251plusmn
01c
273plusmn03a
268plusmn01a
405plusmn003a
397plusmn001a
156plusmn003a
153plusmn003a
200
1287plusmn01cb
1298plusmn01bc
279plusmn01a
277plusmn01a
419plusmn007a
411plusmn004
a15
8plusmn002a
155plusmn
002a
500
1232plusmn02c
1231plusmn01c
222plusmn02c
218plusmn02c
378plusmn006
b370plusmn005b
149plusmn003ab
147plusmn001ab
2DWI
0Mahogany
1532plusmn01b
a1505plusmn01b
213plusmn01c
217plusmn03c
361plusmn004
b367plusmn008b
142plusmn002c
138plusmn001c
501599plusmn01ab
1591plusmn
00a
264plusmn01b
261plusmn01b
397plusmn003a
394plusmn005a
155plusmn004
b14
6plusmn001b
200
1630plusmn01a
1613plusmn03a
268plusmn01b
266plusmn03b
406plusmn006
a401plusmn006
a15
8plusmn008b
149plusmn004
b500
1551plusmn
01b
1527plusmn03b
233plusmn03d
226plusmn01c
373plusmn004
b375plusmn003b
147plusmn001c
141plusmn
001c
6DWI
01441plusmn
02c
1412plusmn03c
255plusmn01b
247plusmn01b
358plusmn002b
368plusmn002b
156plusmn003b
151plusmn
004
b50
1545plusmn01b
1513plusmn01b
299plusmn01a
289plusmn03a
393plusmn001a
395plusmn001a
170plusmn001a
169plusmn002a
200
1555plusmn01b
1525plusmn05b
3 03plusmn01a
297plusmn02a
393plusmn001a
396plusmn004
a174plusmn003a
171plusmn
002a
500
1437plusmn01c
1424plusmn03c
264plusmn01a
258plusmn03b
363plusmn005b
367plusmn002b
159plusmn002b
154plusmn001b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
differentbased
onLSDtest(Ple005)
6 Evidence-Based Complementary and Alternative Medicine
Table3Antioxidant
activ
ityin
leafmethano
licextracts
totalpheno
licandtotalchlorop
hyllcontento
ftwo
HeucheracultivarsV
aluesa
remeans
oftriplicated
eterminationsplusmnsd
Water
interval
Oligosaccharides
treatment
(ppm
)
DPP
Hfre
eradical
scavenging
activ
ity(IC
50120583
gmlminus1
)
120573-C
arotene-lin
oleica
cid
assay
(IC50120583
gmlminus1
)
Totalp
heno
liccontent
(mgGAEgminus1)
Totalchlorop
hyllcontent
(mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
103plusmn001alowast
111plusmn
007a
112plusmn001a
115plusmn001a
104plusmn01c
97plusmn01c
065plusmn004
b063plusmn001bc
5093plusmn006
b99plusmn001b
103plusmn003b
104plusmn002b
109plusmn01b
104plusmn03b
069plusmn002a
067plusmn003a
200
91plusmn005b
99plusmn002b
103plusmn003b
107plusmn003b
108plusmn00b
105plusmn02b
070plusmn002a
068plusmn001a
500
96plusmn003a
104plusmn003a
112plusmn002a
115plusmn004
a104plusmn02c
100plusmn01c
068plusmn001ab
065plusmn002ab
6DWI
082plusmn004
c89plusmn005c
93plusmn001c
101plusmn003c
109plusmn04b
104plusmn02b
061plusmn002c
060plusmn003c
5063plusmn002d
68plusmn000
d74plusmn002d
82plusmn002d
116plusmn03a
112plusmn02a
065plusmn001b
064plusmn002b
200
62plusmn001d
68plusmn001d
72plusmn003d
82plusmn002d
116plusmn01a
113plusmn02a
065plusmn001b
065plusmn003ab
500
78plusmn003c
83plusmn001c
92plusmn004
c99plusmn001c
111plusmn
02b
105plusmn04b
062plusmn002c
061plusmn001c
2DWI
0Mahogany
89plusmn04a
96plusmn01a
101plusmn03a
107plusmn02a
123plusmn02c
117plusmn03c
071plusmn001b
070plusmn002b
5074plusmn002b
79plusmn001b
85plusmn001b
94plusmn003b
127plusmn03b
124plusmn01b
075plusmn002a
074plusmn001a
200
71plusmn001b
78plusmn004
b84plusmn000
b93plusmn003b
129plusmn04b
125plusmn02b
076plusmn001a
075plusmn001a
500
82plusmn002a
88plusmn003a
95plusmn003a
104plusmn002a
122plusmn01c
121plusmn03b
c072plusmn001b
070plusmn002b
6DWI
071plusmn003b
77plusmn002b
84plusmn001b
89plusmn003b
131plusmn01b
124plusmn02b
066plusmn002c
063plusmn001c
5054plusmn001c
57plusmn002c
66plusmn003c
72plusmn003c
138plusmn02a
132plusmn00a
070plusmn001c
068plusmn003c
200
48plusmn00 3c
55plusmn007c
62plusmn002c
71plusmn002c
139plusmn03a
134plusmn01a
071plusmn001a
068plusmn002a
500
68plusmn001b
74plusmn003b
79plusmn003b
82plusmn001b
134plusmn02ab
125plusmn01b
068plusmn002b
064plusmn003b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
different
basedon
LSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
6 Evidence-Based Complementary and Alternative Medicine
Table3Antioxidant
activ
ityin
leafmethano
licextracts
totalpheno
licandtotalchlorop
hyllcontento
ftwo
HeucheracultivarsV
aluesa
remeans
oftriplicated
eterminationsplusmnsd
Water
interval
Oligosaccharides
treatment
(ppm
)
DPP
Hfre
eradical
scavenging
activ
ity(IC
50120583
gmlminus1
)
120573-C
arotene-lin
oleica
cid
assay
(IC50120583
gmlminus1
)
Totalp
heno
liccontent
(mgGAEgminus1)
Totalchlorop
hyllcontent
(mggminus1DW)
2017
2018
2017
2018
2017
2018
2017
2018
2DWI
0Cr
emeB
rulee
103plusmn001alowast
111plusmn
007a
112plusmn001a
115plusmn001a
104plusmn01c
97plusmn01c
065plusmn004
b063plusmn001bc
5093plusmn006
b99plusmn001b
103plusmn003b
104plusmn002b
109plusmn01b
104plusmn03b
069plusmn002a
067plusmn003a
200
91plusmn005b
99plusmn002b
103plusmn003b
107plusmn003b
108plusmn00b
105plusmn02b
070plusmn002a
068plusmn001a
500
96plusmn003a
104plusmn003a
112plusmn002a
115plusmn004
a104plusmn02c
100plusmn01c
068plusmn001ab
065plusmn002ab
6DWI
082plusmn004
c89plusmn005c
93plusmn001c
101plusmn003c
109plusmn04b
104plusmn02b
061plusmn002c
060plusmn003c
5063plusmn002d
68plusmn000
d74plusmn002d
82plusmn002d
116plusmn03a
112plusmn02a
065plusmn001b
064plusmn002b
200
62plusmn001d
68plusmn001d
72plusmn003d
82plusmn002d
116plusmn01a
113plusmn02a
065plusmn001b
065plusmn003ab
500
78plusmn003c
83plusmn001c
92plusmn004
c99plusmn001c
111plusmn
02b
105plusmn04b
062plusmn002c
061plusmn001c
2DWI
0Mahogany
89plusmn04a
96plusmn01a
101plusmn03a
107plusmn02a
123plusmn02c
117plusmn03c
071plusmn001b
070plusmn002b
5074plusmn002b
79plusmn001b
85plusmn001b
94plusmn003b
127plusmn03b
124plusmn01b
075plusmn002a
074plusmn001a
200
71plusmn001b
78plusmn004
b84plusmn000
b93plusmn003b
129plusmn04b
125plusmn02b
076plusmn001a
075plusmn001a
500
82plusmn002a
88plusmn003a
95plusmn003a
104plusmn002a
122plusmn01c
121plusmn03b
c072plusmn001b
070plusmn002b
6DWI
071plusmn003b
77plusmn002b
84plusmn001b
89plusmn003b
131plusmn01b
124plusmn02b
066plusmn002c
063plusmn001c
5054plusmn001c
57plusmn002c
66plusmn003c
72plusmn003c
138plusmn02a
132plusmn00a
070plusmn001c
068plusmn003c
200
48plusmn00 3c
55plusmn007c
62plusmn002c
71plusmn002c
139plusmn03a
134plusmn01a
071plusmn001a
068plusmn002a
500
68plusmn001b
74plusmn003b
79plusmn003b
82plusmn001b
134plusmn02ab
125plusmn01b
068plusmn002b
064plusmn003b
lowastMeans
follo
wedby
different
lette
rswith
incolumns
ares
ignificantly
different
basedon
LSDtest(Ple005)
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
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Hindawiwwwhindawicom Volume 2018
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Disease Markers
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Hindawiwwwhindawicom Volume 2013
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Volume 2018
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ObesityJournal of
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 7
pattern was observed in the second season Furthermorethere was a significant increase in scavenging activity of leafextracts following water stress conditions as revealed bythe 120573-Carotene-linoleic acid assay Heuchera plants (CremeBrulee andMahogany) growing under normal irrigation con-ditions (2DWI) as well as prolonged irrigation (6DWI)showed a significant increase in scavenging activity by leafextracts following application of oligosaccharides at 50 and200 ppm compared to controls and 500 ppm oligosaccharidetreatment in both the 2017 and 2018 years Creme Bruleeplants treated with 200 ppm oligosaccharide showedincreased DPPH (IC
50) free radical scavenging activity in
plants subjected to 2 and 6 days irrigation intervals in the2017 season
Similarly there was a significant increase in total phenoliccontent in plants of both cultivars tested upon widen-ing of the irrigation interval in the two growing seasonsunder study (Table 3) Interestingly oligosaccharide treat-ments boosted phenolic content particularly in plants of bothcultivars treated with 50 and 200 ppm In 2017 Creme Bruleeleaf extracts showed an increase in phenolic content in plantssubjected to 2DWI and 6DWI respectively Similarly thesame year Mahogany leaf extracts showed an increase in phe-nolic content in plants subjected to 2DWI and 6DWI respec-tively Total phenolic content increased significantly in plantstreated with 50 and 200 ppm oligosaccharide compared tothe control and 500 ppm oligosaccharide treatments Totalchlorophyll content in Creme Brulee and Mahogany wassignificantly reduced in control plants subjected to 6DWIIn contrast application of oligosaccharide showed significantincrease in chlorophyll content of treated plants at 50 and200 ppm compared to control and 500 ppm oligosaccharideunder both watering intervals in both cultivars and inthe two growth seasons evaluated In summary antioxidantactivity and phenolic and chlorophyll contents were higherinMahogany than in Creme Brulee in the two seasons understudy
33 Enzymatic and Nonenzymatic Antioxidants Majorantioxidant SOD CAT and APX enzyme activities showedsignificant increases in Creme Brulee and Mahogany plantssubjected to oligosaccharide treatments at 50 and 200 ppmcompared to oligosaccharides at 500 ppm and controltreatments under normal and prolonged irrigation intervals(Figure 1) In both cultivars application of oligosaccharide at200 ppm resulted in the highest SOD CAT and APX enzymeactivities recorded both under 2DWI and 6DWI and in bothseasons studied Mahogany plants showed slightly highervalues of SOD CAT and APX enzymes activities comparedto Creme Brulee
Free and total ascorbate (nonenzymatic antioxidants)showed a significant increase in oligosaccharides-treatedplants at 50 and 200 ppm compared to oligosaccharide at 500ppm and control treatments under normal and prolongedirrigation intervals (Figure 2) Concomitantly there weresignificant reductions in H
2O2content in oligosaccharides-
treated plants at 50 and 200 ppm compared to the 500 ppmdose as well as the control treatment in both cultivars and inboth seasons (Figure 2)
025
02
015
01
005
02DWI Brulee 6DWI
Mahogany2DWI Brulee 6DWI
Mahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aab b cc
abc
aab c
c
c
c c
SOD
activ
ity (U
nit m
g-1
prot
ein)
CAT
activ
ity (
mol
g-1
pro
tein
)A
PX ac
tivity
(m
ol g
-1 p
rote
in)
0
01
02
03
04
05
06
07
08
09
0
1
2
3
4
5
6
7
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
aaaa
b
b
aaaa b
bb
bb
aaa
ab b
baa
bb
aabb
c
Figure 1 SOD CAT and APX activities in Heuchera subjected toprolonged irrigation intervals and different oligosaccharides (OL)concentrations
34 Antibacterial and Antifungal Activities Heuchera CremeBrulee leaf extracts showed antibacterial activities againstscreened bacteria as shown in Table 4 The highest antibac-terial activities were found in plants subjected to prolongedirrigation intervals and 200500 ppm oligosaccharide InMahogany plants there were higher antibacterial activitiesof leaf extracts against the same collection of bacteria Thehighest antibacterial activities were against B cereus and Mflavus in plants treated with prolonged irrigation intervalsand 500 ppm oligosaccharide Both cultivars leaf extractsshowed comparable antibacterial activities to antibioticsunder stress and oligosaccharides treatments
The antifungal activities of Heuchera cultivars leafextracts were investigated as shown in Table 5 Creme Bruleeshowed antifungal activities as well as Mahogany In both
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
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Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
8 Evidence-Based Complementary and Alternative Medicine
Table4Minim
uminhibitory
(MIC)a
ndbactericidalconcentration(M
BC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(m
gminus1mL)
forthe
2018
grow
ingseason
Water
interval
Oligosaccharides
treatment(pp
m)
Escherich
iacoli
Staphylococcus
aureus
Bacillus
cereus
Micr
ococcus
flavus
Pseudomonas
aerugin
osa
Liste
riamonocyto-
genes
2DWI
0Cr
emeB
rulee
023plusmn001
014plusmn001
010plusmn002
011plusmn001
013plusmn002
020plusmn001
045plusmn001
033plusmn003
021plusmn001
022plusmn002
027plusmn001
040plusmn001
200
021plusmn003
013plusmn002
09plusmn004
010plusmn001
012plusmn001
019plusmn002
042plusmn001
031plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
500
019plusmn005
012plusmn003
08plusmn002
09plusmn003
011plusmn002
018plusmn001
040plusmn001
030plusmn001
017plusmn001
019plusmn001
023plusmn001
035plusmn001
6DWI
0020plusmn005
012plusmn006
09plusmn001
010plusmn001
011plusmn001
019plusmn002
040plusmn001
029plusmn001
018plusmn001
020plusmn001
024plusmn001
037plusmn001
200
01 8plusmn001
011plusmn001
07plusmn003
09plusmn002
010plusmn001
017plusmn002
039plusmn001
027plusmn001
017plusmn001
019plusmn001
021plusmn001
033plusmn001
500
017plusmn001
010plusmn004
06plusmn004
08plusmn001
09plusmn002
015plusmn004
038plusmn001
023plusmn001
015plusmn001
016plusmn000
018plusmn001
030plusmn001
2DWI
0Mahogany
020plusmn04
012plusmn03
09plusmn03
010plusmn001
011plusmn002
017plusmn003
040plusmn001
028plusmn001
017plusmn001
020plusmn001
024plusmn001
033plusmn001
200
018plusmn001
011plusmn004
08plusmn000
09plusmn003
010plusmn002
016plusmn002
038plusmn001
025plusmn001
016plusmn001
019plusmn001
021plusmn001
031plusmn001
500
017plusmn001
810plusmn003
07plusmn003
08plusmn002
009plusmn001
015plusmn003
036plusmn001
023plusmn001
014plusmn001
016plusmn001
018plusmn001
030plusmn001
6DWI
0018plusmn001
010plusmn002
08plusmn001
08plusmn004
010plusmn001
016plusmn002
038plusmn001
023plusmn001
016plusmn001
016plusmn001
020plusmn001
031plusmn001
200
01 6plusmn003
09plusmn007
07plusmn002
07plusmn002
09plusmn003
015plusmn001
035plusmn001
020plusmn001
014plusmn001
014plusmn002
018plusmn001
030plusmn001
500
015plusmn001
07plusmn003
05plusmn003
06plusmn001
08plusmn002
013plusmn001
033plusmn001
018plusmn001
012plusmn001
012plusmn001
016plusmn001
027plusmn001
Streptom
ycin
09plusmn001
020plusmn001
005plusmn001
010plusmn000
5007plusmn000
016plusmn001
042plusmn001
043plusmn001
014plusmn001
019plusmn000
5014plusmn001
033plusmn001
Ampicillin
024plusmn001
010plusmn003
010plusmn000
5010plusmn0002
014plusmn001
016plusmn001
044plusmn001
015plusmn001
018plusmn000
5016plusmn000
5022plusmn001
028plusmn001
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
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MEDIATORSINFLAMMATION
of
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Hindawiwwwhindawicom Volume 2018
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Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
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Oxidative Medicine and Cellular Longevity
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PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
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Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
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OphthalmologyJournal of
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Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
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Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 9
Table5Minim
uminhibitory
(MIC)and
fung
icidalconcentration(M
FC)o
fHeucheraCr
emeB
ruleea
ndMahoganyleafextracts(mgminus1mL)
Water
interval
Oligosaccharides
treatment(pp
m)
Aspergillus
niger
MIC
MFC
Aspergillus
ochraceus
MIC
MFC
Aspergillus
flavus
MIC
MFC
Penicilliu
mochrochloron
MIC
MFC
Cand
ida
albicans
MIC
MFC
2DWI
0Cr
emeB
rulee
020plusmn001
021plusmn001
013plusmn002
025plusmn001
014plusmn002
042plusmn001
043plusmn003
027plusmn001
053plusmn002
027plusmn001
200
020plusmn003
019plusmn002
012plusmn001
023plusmn001
012plusmn001
041plusmn001
040plusmn001
025plusmn001
050plusmn001
024plusmn001
500
019plusmn003
017plusmn003
011plusmn002
021plusmn003
011plusmn002
040plusmn001
035plusmn001
023plusmn001
048plusmn001
023plusmn001
6DWI
0018plusmn005
018plusmn001
012plusmn001
022plusmn001
011plusmn001
039plusmn001
037plusmn001
026plusmn001
049plusmn001
024plusmn001
200
016plusmn001
017plusmn001
011plusmn001
020plusmn002
010plusmn001
035plusmn001
036plusmn001
022plusmn001
045plusmn001
021plusmn001
500
015plusmn001
015plusmn003
010plusmn001
019plusmn001
09plusmn002
033plusmn001
033plusmn001
021plusmn001
043plusmn001
018plusmn001
2DWI
0Mahogany
017plusmn001
016plusmn03
012plusmn001
021plusmn001
011plusmn002
033plusmn001
036plusmn001
025plusmn001
044plusmn001
024plusmn001
200
016plusmn001
015plusmn002
011plusmn000
020plusmn003
010plusmn002
031plusmn001
034plusmn001
026plusmn001
041plusmn001
021plusmn001
500
015plusmn001
814plusmn003
010plusmn003
019plusmn002
009plusmn001
030plusmn001
029plusmn001
020plusmn001
039plusmn001
018plusmn001
6DWI
0016plusmn001
015plusmn002
011plusmn001
020plusmn004
010plusmn001
032plusmn001
032plusmn001
025plusmn001
040plusmn001
020plusmn001
200
014plusmn003
013plusmn001
010plusmn002
019plusmn002
09plusmn003
030plusmn001
027plusmn001
020plusmn001
038plusmn00 2
018plusmn001
500
012plusmn001
012plusmn003
09plusmn003
017plusmn001
08plusmn002
025plusmn001
025plusmn001
019plusmn001
035plusmn001
016plusmn001
FLZ
015plusmn001
020plusmn001
013plusmn001
021plusmn001
010plusmn001
028plusmn003
033plusmn001
022plusmn003
033plusmn001
021plusmn001
KTZ
010plusmn001
021plusmn001
021plusmn001
019plusmn001
020plusmn001
020plusmn001
040plusmn001
040plusmn001
042plusmn001
040plusmn001
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
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Hindawiwwwhindawicom Volume 2018
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Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
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Oxidative Medicine and Cellular Longevity
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PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
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Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
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Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
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OphthalmologyJournal of
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Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
10 Evidence-Based Complementary and Alternative MedicineFr
ee as
corb
ate (
To
tal a
scor
bate
(m
ol g
-1 D
W)
H2
O2
cont
ent (
m
ol g
-1 D
W)
0
005
01
015
02
025
03
035
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
50
100
150
200
250
300
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
0
20
40
60
80
100
120
2DWI Brulee 6DWIMahogany
2DWI Brulee 6DWIMahogany
ControlOL 50 ppm
OL 200 ppmOL 500 ppm
a a a a a a a a
aaa
a a a a a
abab ab
bb
b b b b b b c b
bbc
a aa ab a ab
a abb bb bb b
b b
m
ol g
-1 D
W)
Figure 2 Free and total ascorbate and H2O2content in Heuchera
plants subjected to prolonged irrigation intervals and differentoligosaccharides (OL) concentrations
cultivars prolonged irrigation and oligosaccharide treat-ments (500 and 200ppm) showed the highest antifungalactivities The antifungal activities of Mahogany leaf extractswere higher than Creme Brulee and were comparable toantibiotics
4 Discussion
A significant reduction in morphological parameters such asplant height number of leaves leaf area and plant dryweightdue to extension of the irrigation interval which is in agree-ment with previous studies [20 40ndash42]Thesemorphologicalchanges associated with major physiological alterations suchas changes in carbohydrate K Ca proline chlorophylls andantioxidants contents [15 21 42] Oligosaccharide sprays at
specific doses enhanced the growth of the two Heucheracultivars tested here during normal and extended irrigationintervals as reflected by increased vegetative growth Similarobservations have been described before for oligosaccharidetreatments on dry matter and essential oil yield in ThymusdaenensisCelak [28] In that study the authors suggested thatthe increase in dry matter and in the essential oil yield undermild stress might be attributed to increased proline contentand to lipid peroxidation
Accumulation of carbohydrates might be an importantindicator of stress tolerance in plants by means of osmoticadjustment and scavenging of ROS [43 44] Additionallythe accumulation of proline balances vacuolar ion osmoticpressure [20 40] and maintains water influx [45] Prolineaccumulation increased under an extended irrigation intervalin the present study an original contribution of the studyreported herein is that we report the increase in leaf prolinecontent at normal irrigation interval something not previ-ously reported using low doses of 50 and 200 ppm oligosac-charideThe accumulation of K andCa ions in plant leaves is awell-known mechanism of osmotic adjustment during stressconditions such as drought and salinity This accumulationof K and Ca is associated with carbohydrate accumula-tion in stressed plants which enhances plant performanceduring stress and improves cell turgor pressure [21 40]Interestingly K and Ca accumulation in plant during stressconditions enhance photosynthetic rate leading to increasedchlorophyll content (drought resistance mechanism) as wellas carbohydrate accumulation such as documented hereinwhich helped in improving plant performance during stressThe application of oligosaccharide at low rate significantlyincreased leaf K and Ca content and helped in attainingosmotic adjustment during water stress Such accumulationof K and Ca in plants might be associated with antifungalactivities [46ndash48]
Excess ROS eg H2O2 O2 and OHminus are produced
in plants under water stress conditions due to imbalancebetween production and utilization of electrons This condi-tion may cause damage and even cell death [49] if ROS arenot effectively removed An antioxidant defense mechanismin plants consists of enzymatic and nonenzymatic tools thatintervene to maintain the intracellular redox balance underconditions of stress Nonenzymatic tools include secondarymetabolites such as total and free ascorbate as well as phe-nols and their derivatives (eg flavanones and anthocyanins)[21 50 51] Enzymatic tools include many enzymes amongwhich the most common are SOD CAT and APX whichcontrol H
2O2production in plants [44 50] Further these
compounds including ascorbate (derivative of ascorbic acid)have well-known antibacterial and antifungal activities asfound in this study [52ndash55] In the current study we foundstrong antibacterial and antifungal activities in plants withaccumulated ascorbate as in plants subjected to prolongedand oligosaccharide treatments
We observed a significant increase in leaves phenoliccomposition following water stress conditions which becamehigher in oligosaccharides-treated plants This increase intotal phenolic content in leaves was reflected in an increasein antioxidant activity as determined by the DPPH and
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 11
linoleic acid assays Additionally oligosaccharide doses of50 and 200 ppm caused a significant increase in phenoliccontent in leaves compared to control plants these resultsare consistent with those reported by [26] who reportedhigher polyphenols in plants of Origanum vulgare subjectedto oligosaccharide treatment Phenols play an important rolein removing ROS in stressed plants and largely affect theantioxidant estimations of DPPH and linoleic acid assayswhich mainly measure OHminus free radical The higher antibac-terial and antifungal activities found in both cultivars leafextracts in plants subjected to prolonged irrigation intervalsand oligosaccharide treatments are associated with the accu-mulation of phenols in treated plants The accumulation ofphenols has important inhibiting effects on the growing ofbacteria and fungi [56 57] It was also clear that CremeBruleeand Mahogany differed from one another with respect tomorphological (eg plant height) physiological parameters(eg phenolic composition and antioxidant) and biochem-ical activities (antibacterial and antifungal activities) Suchvariation between cultivars has been documented for otherspecies in response to stress and genetics [58 59]
5 Conclusion
This is the first report on enhancing Heuchera plants medic-inal values by subjecting the plants to water stress andoligosaccharide treatments Morphological physiologicaland antimicrobial parameters were studied to documentthe interaction between stress tolerance and oligosaccha-ride applications in Heuchera plants subjected to waterstress and oligosaccharide treatments The study revealedthat oligosaccharide foliar application effectively amelioratedwater stress deleterious effects on plant growth enhancedthe phytochemical composition and improved themedicinalvalues of treated plants These findings suggest that waterstress accompanied by oligosaccharide sprays might be avaluable tool in improving the medicinal value in horti-cultural crops and the future development of novel toolsto control food borne pathogens and respective microbialdiseases
Data Availability
All data used to support the findings of this study are includedwithin the article
Conflicts of Interest
The authors declare that they have no conflicts of interest
Authorsrsquo Contributions
Hosam O Elansary Eman A Mahmoud and Amal M EAbdel-Hamid mainly designed the study and performedexperiments Fahed A Al-Mana Diaa O Elansary and TarekK Ali Zin El-Abedin were responsible mainly for fundingacquisition data analyses and final presentation All theauthors participated in the analyses writing revising andapproving the final version of the manuscript
Acknowledgments
The authors extend their appreciation to the Deanship ofScientific Research at King Saud University for funding thiswork through Research Group no RG-1440-12
References
[1] J L Schoeni and A C Lee Wong ldquoBacillus cereus food poi-soning and its toxinsrdquo Journal of Food Protection vol 68 no 3pp 636ndash648 2005
[2] T Li D Zhang T N Oo et al ldquoInvestigation on the antibac-terial and anti-T3SS Activity of traditional myanmar medic-inal plantsrdquo Evidence-Based Complementary and AlternativeMedicine vol 2018 Article ID 2812908 13 pages 2018
[3] W Van Schaik M H Zwietering W M De Vos and TAbee ldquoDeletion of the sigB gene in Bacillus cereus ATCC14579 leads to hydrogen peroxide hyperresistancerdquo Applied andEnvironmental Microbiology vol 71 no 10 pp 6427ndash64302005
[4] JM Farber and P I Peterkin ldquoListeria monocytogenes a food-borne pathogenrdquoMicrobiology Reviews vol 55 no 3 pp 476ndash511 1991
[5] V Toledo H C Den Bakker J C Hormazabal et al ldquoGenomicdiversity of listeria monocytogenes isolated from clinical andnon-clinical samples in Chilerdquo Gene vol 9 no 8 p 396 2018
[6] E M Mateo F M Valle-Algarra R Mateo-Castro and MJimenez ldquoImpact of non-selective fungicides on the growthand production of ochratoxin a by aspergillus ochraceus andA carbonarius in barley-based mediumrdquo Food Additives andContaminants - Part A Chemistry Analysis Control Exposureand Risk Assessment vol 28 no 1 pp 86ndash97 2011
[7] U B Kakde andHU Kakde ldquoIncidence of post-harvest diseaseand airborne fungal spores in a vegetablemarketrdquoActa BotanicaCroatica vol 71 no 1 pp 147ndash157 2012
[8] N Gemeda A Tadele H Lemma et al ldquoDevelopment charac-terization and evaluation of novel broad-spectrum antimicro-bial topical formulations fromrdquoEvidence-BasedComplementaryandAlternativeMedicine vol 2018 Article ID 9812093 16 pages2018
[9] J W M Van Der Linden A Warris and P E Verweij ldquoAsper-gillus species intrinsically resistant to antifungal agentsrdquoMedi-cal Mycology vol 49 no 1 pp S82ndashS89 2011
[10] M Canica V Manageiro H Abriouel J Moran-Giladde andC M A P Franz ldquoAntibiotic resistance in foodborne bacteriardquoTrends in Food Science amp Technology
[11] J R Beddington M Asaduzzaman M E Clark et al ldquoThe rolefor scientists in tackling food insecurity and climate changerdquoAgriculture amp Food Security vol 2012 pp 1ndash10 2012
[12] G Ondrasek ldquoWater scarcity and water stress in agriculturerdquoin Physiological Mechanisms and Adaptation Strategies in Plantsunder Changing Environment P Ahmad and M R Wani Edspp 74-75 Springer New York NY USA 2014
[13] B Pallas A Clement-Vidal M-C Rebolledo J-C Soulie andD Luquet ldquoUsing plant growth modeling to analyze C source-sink relations under drought Inter- and intraspecific compari-sonrdquo Frontiers in Plant Science vol 4 p 437 2013
[14] S Hussain F Khan W Cao L Wu and M Geng ldquoSeed prim-ing alters the production and detoxification of reactive oxygenintermediates in rice seedlings grown under sub-optimal tem-perature and nutrient supplyrdquo Frontiers in Plant Science vol 7p 439 2016
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
12 Evidence-Based Complementary and Alternative Medicine
[15] M A El-Esawi H O Elansary N A El-Shanhorey AM Abdel-Hamid H M Ali and M S Elshikh ldquoSalicylicacid-regulated antioxidant mechanisms and gene expressionenhance rosemary performance under saline conditionsrdquo Fron-tiers in Physiology vol 8 p 716 2017
[16] O Tebboub R Cotugno F Oke-Altuntas et al ldquoAntioxi-dant potential of herbal preparations and components fromGalactites elegans (All) Nyman ex Soldanordquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID9294358 7 pages 2018
[17] A Muniz E Garcia D Gonzalez and L Zuniga ldquoantioxidantactivity and in vitro antiglycation of the fruit of Spondias pur-pureardquo Evidence-Based Complementary and Alternative Medi-cine vol 2018 Article ID 5613704 7 pages 2018
[18] C J Atkinson J D Fitzgerald and N A Hipps ldquoPotentialmechanisms for achieving agricultural benefits from biocharapplication to temperate soils a reviewrdquo Plant and Soil vol 337no 1 pp 1ndash18 2010
[19] A Sos-Hegedus Z Juhasz P Poor et al ldquoSoil drench treat-ment with szlig-aminobutyric acid increases drought tolerance ofpotatordquo PLoS ONE vol 9 no 12 p e114297 2014
[20] H O Elansary and M Z M Salem ldquoMorphological andphysiological responses and drought resistance enhancementof ornamental shrubs by trinexapac-ethyl applicationrdquo ScientiaHorticulturae vol 189 pp 1ndash11 2015
[21] H O Elansary K Yessoufou A M E Abdel-Hamid M A El-Esawi H Ali and M S Elshikh ldquoSeaweed extracts enhancesalam turfgrass performance during prolonged irrigation inter-vals and saline shockrdquo Frontiers in Plant Science vol 8 p 8302017
[22] R Saxena R S Tomar andM Kumar ldquoExploring nanobiotech-nology to mitigate abiotic stress in crop plantsrdquo Journal ofPharmaceutical Sciences and Research vol 8 no 9 pp 974ndash9802015
[23] R Sharp ldquoA review of the applications of chitin and its deriva-tives in agriculture to modify plant-microbial interactions andimprove crop yieldsrdquo Journal of Agronomy vol 3 no 4 pp 757ndash793 2013
[24] R Pichyangkura and S Chadchawan ldquoBiostimulant activity ofchitosan in horticulturerdquo Scientia Horticulturae vol 196 pp49ndash65 2015
[25] S Karuppusamy ldquoA review on trends in production of sec-ondary metabolites from higher plants by in vitro tissue organand cell culturesrdquo Journal of Medicinal Plants Research vol 3no 13 pp 1222ndash1239 2009
[26] H Yin X C Frette L P Christensen and K Grevsen ldquoChito-san oligosaccharides promote the content of polyphenols inGreek oregano (Origanum vulgare ssp hirtum)rdquo Journal ofAgricultural and FoodChemistry vol 60 no 1 pp 136ndash143 2012
[27] M A Ramırez A T Rodriguez L Alfonso and C PenicheldquoChitin and its derivatives as biopolymers with potential agri-cultural applicationsrdquo Biotecnologıa Aplicada vol 27 pp 270ndash276 2010
[28] Z Emami Bistgani S A Siadat A Bakhshandeh A GhasemiPirbalouti and M Hashemi ldquoInteractive effects of droughtstress and chitosan application on physiological characteristicsand essential oil yield ofThymus daenensis CelakrdquoCrop Journalvol 5 no 5 pp 407ndash415 2017
[29] R A Folk J RMandel and J V F Reudenstein ldquoAncestral geneflow and parallel organellar genome capture result in extremephylogenomic discord in a lineage of angiospermsrdquo SystematicBiology vol 66 no 3 pp 320ndash337 2017
[30] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland MA USA 2nd edition 2002
[31] D E Soltis ldquoSaxifragaceaerdquo inThe Families and Genera of Vas-cular Plants K Kubitzki Ed vol 9 p 422 2007
[32] M Dubois K A Gilles J K Hamilton P A Rebers and FSmith ldquoColorimetric method for determination of sugars andrelated substancesrdquoAnalytical Chemistry vol 28 no 3 pp 350ndash356 1956
[33] Y Jiang and B Huang ldquoOsmotic adjustment and root growthassociated with drought preconditioning-enhanced heat toler-ance in Kentucky bluegrassrdquo Crop Science vol 41 no 4 pp1168ndash1173 2001
[34] W A Torello and L A Rice ldquoEffects of NaCl stress on pro-line and cation accumulation in salt sensitive and tolerant turf-grassesrdquo Plant and Soil vol 93 no 2 pp 241ndash247 1986
[35] L S Bates R P Waldren and I D Teare ldquoRapid determinationof free proline for water-stress studiesrdquo Plant and Soil vol 39no 1 pp 205ndash207 1973
[36] M A Amerine and C S Ough ldquoPhenolic compoundsrdquo inMethods for Analysis of Musts and Wines pp 196ndash219 JohnWiley and Sons New York NY USA 1988
[37] V L Singleton and J A Rossi ldquoColorimetry of total phenolicswith phosphomolybdic-phosphotungistic acid reagentsrdquoAmer-ican Journal of Enology andViticulture vol 16 pp 144ndash158 1965
[38] R Moran and D Porath ldquoChlorophyll determination in intacttissues using NN-dimethylformamiderdquo Plant Physiology vol65 no 3 pp 478-479 1980
[39] H O Elansary A Szopa P Kubica et al ldquoBioactivities oftraditional medicinal plants in Alexandriardquo Evidence-BasedComplementary and Alternative Medicine vol 2018 Article ID1463579 13 pages 2018
[40] F Ali A Bano and A Fazal ldquoRecent methods of drought stresstolerance in plantsrdquo Plant Growth Regulation vol 82 no 3 pp363ndash375 2017
[41] C G Goncalves A C da Silva Junior M R R Pereira E CGasparino and D Martins ldquoMorphological modifications insoybean in response to soil water managementrdquo Plant GrowthRegulation vol 83 no 1 pp 105ndash117 2017
[42] M Caser C Lovisolo and V Scariot ldquoThe influence of waterstress on growth ecophysiology and ornamental quality ofpotted Primula vulgaris lsquoHeidyrsquo plants New insights to increasewater use efficiency in plant productionrdquo Plant Growth Regula-tion vol 83 no 3 pp 361ndash373 2017
[43] Y-G Yin Y Kobayashi A Sanuki et al ldquoSalinity induces carbo-hydrate accumulation and sugar-regulated starch biosyntheticgenes in tomato (Solanum lycopersicum L cv rsquoMicro-Tomrsquo)fruits in an ABA-and osmotic stress-independent mannerrdquoJournal of Experimental Botany vol 61 no 2 pp 563ndash574 2010
[44] B Gupta and B Huang ldquoMechanism of salinity tolerance inplants physiological biochemical and molecular characteriza-tionrdquo International Journal of Genomics vol 2014 Article ID701596 18 pages 2014
[45] M A Hoque M N A Banu E Okuma et al ldquoExogenousproline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salttolerance more than glycinebetaine in tobacco Bright Yellow-2suspension-cultured cellsrdquo Journal of Plant Physiology vol 164no 11 pp 1457ndash1468 2007
[46] F-F Liu L Pu Q-Q Zheng et al ldquoCalcium signaling mediatesantifungal activity of triazole drugs in the Aspergillirdquo FungalGenetics and Biology vol 81 pp 182ndash190 2014
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Evidence-Based Complementary and Alternative Medicine 13
[47] H Jabnoun-Khiareddine R Abdallah R El-Mohamedy et alldquoComparative efficacy of potassium salts against soil-borne andair-borne fungi and their ability to suppress tomato wilt andfruit rotsrdquo Journal of Microbiology and Biochemical Technologyvol 8 pp 045ndash055 2016
[48] W Lee and D G Lee ldquoPotential role of potassium andchloride channels in regulation of silymarin-induced apoptosisin Candida albicansrdquo IUBMB Life vol 70 no 3 pp 197ndash2062018
[49] C M Cruz ldquoDrought stress and reactive oxygen species pro-duction scavenging and signalingrdquo Plant Signaling and Behav-ior vol 3 pp 156ndash165 2008
[50] Y Fang and L Xiong ldquoGeneral mechanisms of droughtresponse and their application in drought resistance improve-ment in plantsrdquo Cellular andMolecular Life Sciences vol 72 no4 pp 673ndash689 2015
[51] H AbdElgawad G Zinta M M Hegab R Pandey H Asardand W Abuelsoud ldquoHigh salinity induces different oxidativestress and antioxidant responses in maize seedlings organsrdquoFrontiers in Plant Science vol 7 p 276 2016
[52] A Belicova J Dobias L Ebringer and J Krajcovic ldquoEffect ofascorbic acid on the antimicrobial activity of selected antibioticsand synthetic chemotherapeutical agents in the in vitro condi-tionsrdquo Ceska a Slovenska Farmacie vol 49 no 3 pp 134ndash1382000
[53] M Tajkarimi and S A Ibrahim ldquoAntimicrobial activity ofascorbic acid alone or in combination with lactic acid onEscherichia coli O157H7 in laboratory medium and carrotjuicerdquo Food Control vol 22 no 6 pp 801ndash804 2011
[54] F Van Hauwenhuyse A Fiori P Van Dijck F Van Hauwen-huyse A Fiori and P Van Dijck ldquoAscorbic acid inhibition ofcandida albicans Hsp90-mediated morphogenesis occurs viathe transcriptional regulator Upc2rdquo Eukaryotic Cell vol 13 no10 pp 1278ndash1289 2014
[55] P Avci F Freire A Banvolgyi E Mylonakis N M Wikonkaland M R Hamblin ldquoSodium ascorbate kills Candida albicansin vitro via iron-catalyzed Fenton reaction Importance ofoxygenation and metabolismrdquo Future Microbiology vol 11 no12 pp 1535ndash1547 2016
[56] A Rao Y Zhang S Muend and R Rao ldquoMechanism of anti-fungal activity of terpenoid phenols resembles calcium stressand inhibition of the TOR pathwayrdquo Antimicrobial Agents andChemotherapy vol 54 no 12 pp 5062ndash5069 2010
[57] R P Pizzolitto C L Barberis J S Dambolena et al ldquoInhibitoreffect of natural phenolic compounds onAspergillus parasiticusgrowthrdquo Journal of Chemistry vol 2015 Article ID 547925 7pages 2015
[58] H O Elansary and E A Mahmoud ldquoBasil cultivar identifi-cation using chemotyping still favored over genotyping usingcore barcodes and possible resources of antioxidantsrdquo Journalof Essential Oil Research vol 27 no 1 pp 82ndash87 2015
[59] H O Elansary and K Yessoufou ldquoIn vitro antioxidant antifun-gal and antibacterial activities of five international Calibrachoacultivarsrdquo Natural Product Research (Formerly Natural ProductLetters) vol 30 no 11 pp 1339ndash1342 2016
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
Stem Cells International
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
MEDIATORSINFLAMMATION
of
EndocrinologyInternational Journal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Disease Markers
Hindawiwwwhindawicom Volume 2018
BioMed Research International
OncologyJournal of
Hindawiwwwhindawicom Volume 2013
Hindawiwwwhindawicom Volume 2018
Oxidative Medicine and Cellular Longevity
Hindawiwwwhindawicom Volume 2018
PPAR Research
Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom
The Scientific World Journal
Volume 2018
Immunology ResearchHindawiwwwhindawicom Volume 2018
Journal of
ObesityJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Computational and Mathematical Methods in Medicine
Hindawiwwwhindawicom Volume 2018
Behavioural Neurology
OphthalmologyJournal of
Hindawiwwwhindawicom Volume 2018
Diabetes ResearchJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
Research and TreatmentAIDS
Hindawiwwwhindawicom Volume 2018
Gastroenterology Research and Practice
Hindawiwwwhindawicom Volume 2018
Parkinsonrsquos Disease
Evidence-Based Complementary andAlternative Medicine
Volume 2018Hindawiwwwhindawicom
Submit your manuscripts atwwwhindawicom
