Effect of Water Stress on Performance of Maize Inoculated With Glomus Sp

8/14/2019 Effect of Water Stress on Performance of Maize Inoculated With Glomus Sp

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9/21/13 Effect of water stress on performance of maize inoculated with Glomus sp

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Effect of water stress on performance of maize inoculated with Glomussp. isolated from tea garden o

Keonjhar , Orissa

N. Gupta* and S. Routaray

Microbiology Laboratory, Division of Biotechnology

Regional Plant Resource Centre, Bhubaneswar 751 015, Orissa , India* Corresponding Author Email :[email protected]

Abstract :

A study was conducted to determine the effects of arbuscular mycorrhizal (AM) fungi inoculation on growth omaize grown under well watered and water stressed pot culture conditions. 15 days old maize seedlings were

planted in earthen pots after treatment with or without the AM fungi. Roots were sampled after 75 days ofgrowth stages to quantify AM fungi. Mycorrhizal colonization was higher in water-stressed plants than well-watered plants. Biomass was higher in mycorrhizal than nonmycorrhizal plants irrespective of water treatments.However the plants irrigated with alternate watering schedule had shown higher biomass and than those treated

with daily watering. The improved growth in maize plants reported here demonstrate the potential of mycorrhizainoculation to reduce the effects of drought stress and useful for the better performance under stress conditions.

Key words : Mycorrhiza, stress, drought, tea, Glomus

Introduction:

Many microorganisms including the AM fungi facilitate uptake of mineral nutrients from soil.

Enhancement of growth and yield of the host plants has been reported due to AM inoculations (Freitas et al.

2004 ). These fungi not only play a vital role in uptake kinetics of the host plants but also act as carrier and

being symbiont, use to transport the nutrients into the host plant roots ( Gupta and Baig, 2001, VogelMikus et al.

2005). It is also worthwhile to note that the AM fungi help the host plants to grow under stressed environment

(Mc Millen et al., 1998; Thaker and Jasrai, 2002, Gupta and Routaray, 2005). Water stress is the majo

edaphic limiting factor, which affects establishment and efficiency of mycorrhiza grown in association with various

crops (AlKaraki et al., 2004).

Mycorrhizal benefits in plant mineral nutrition are being increasingly recognized in forestry, agriculture and

plantation crops. Tea (Camellia sinensis (L.) O. Kuntz ), one of the major plantation crops, earn

considerable amount of foreign exchange and provides avenues for employment. As most tropical plants seem

to be obligatory mycotrophic, there is increasing interest to strengthen the technology for better tea cultivation

Although knowledge on the establishment of mycorrhiza with the tea soil is in its infancy, there is a need to

understand the AM spores occurrence and the dominant species associated with tea. Realizing the importance o

AM technology in tea production; this study was under taken on the AM fungi indigenous to the regions for the

first time ever. The present study attains more importance due to the study sites that has been taken into

consideration. The tea gardens surveyed were drought prone providing scope to isolate stress tolerant strains o

AM fungi. Bhuyanpirh tea plantations were never surveyed before. Therefore, the present study is a kind of firs

report that increases the possible occurrence of the new strains of AM fungi.


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Materials and Methods

Study site and collection of soil samples

The soil and root samples were brought from the tea (Camelia sinensisL.) plantations of the Bhuyanpirh tea

estate of M/S Orissa Tea Plantation Limited situated in Tarmakanta about 48 Km away from Keonjhar city o

Orissa, India. The plantation was placed in an area which was once covered by dry and mixed deciduous sa

forests at an elevation of more than 600 m. The soil was red clay-loam and poor in nutrient content. Thrhizosphere soil of different tea plants was collected from the 10 inch below ground in polythene bags and

brought for analysis.

Isolation , identification and pure culture of AM fungal spores

The isolation of spore were carried out following the method of Gerdemann and Nicolson (1963). Th

representative soil samples (100 g) from each location (in triplicate) was suspended in sufficient quantity of wate

and stirred thoroughly. Resulting soil suspension was sieved through mesh sizes 400, 300, 200, and 100 m

placed one below the other in the same order. The residues left after sieving were filtered through whatmanpaper no. 1 and observed under stereo zoom microscope ( Meiji, Japan) for spore count.The isolated spore

were given a thorough microscopic examination to record their morpho-taxonomic features and tentative

identification was made (Schenck and Perez, 1987) .AM spores obtained from the tea plantation rhizosphere

soils were purified following the funnel technique (Menge and Timmer , 1982) for which single spore isolation

was done by picking AM spores from the spore mass left on the sieves. A device was made by keeping a funne

on the bottle (filled with nutrient solution). This funnel was first filled with small amount of sterilized sand and soi

mix (1:1) up to the neck portion. At the neck portion of funnel, individual spore of AM fungi was kept and filled

with sand-soil mix. Maize seeds were sown under glass house and their roots were allowed to come into contacwith spore through neck portion. During the growing for one month period, daily watering was done and weekly

nutrient solution (1/4 strength Hoagland) was added. Afterwards, seedling roots were analyzed for colonization

of AM fungi. The complete system including soil and seedlings were transferred into the bigger earthen po

containing sterilized sand soil and soilrite mix for the multiplication of individual spores. The pure culture o

isolated AM fungi were used for pot culture inoculations of rice, onion and maize as the hosts for their

multiplication. During multiplication, host plants were nourished by hoagland nutrient solution quarterly and daily

watering was done up to three months. The pure culture of this AM fungi was used for futher inoculation into the

experimental sets.

Experimental set up

Experiment was set with sterilized Black cotton soil in mixed with sand and compost under glasshouse

conditions. Maize Seedlings were raised in the earthen pots having 5-Kg capacity containing soil sand mixture

(1;1) as growing media irrigated with fresh water and cultivated under glass house conditions at 32 2

C and 80 5 % relative humidity. At the beginning of the experiment, the values for total dry weigh

biomass of the seedlings by oven drying at 80 C and values for average leaf area per plant were measured


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and total of 40 uniform and 15 days old seedlings of maize were distributed in to four treatments groups i. e. 10

seedlings of each group were sub irrigated according to following experimental protocol.

Experimental group I: only soil (Black cotton soil +sand + compost , 2:1:1) in 5 Kg earthern pots + maize plants

+ daily watering

Experimental group II: soil inoculated with AM fungi (500 600 spores pot) + maize plants +daily watering

(250 ml)

Experimental group III. Soil inoculated with AM fungi + 15 day-old- maize seedlings treated with followingsub treatments for 60 days of growth period. Watering schedule Watering was done in two ways. Firstly

every alternate day and then at two days interval. For the rest of the experimental sets daily watering wa

maintained.

In this experiment the maize plants were uprooted and measured for the growth parameters at an interval of 30

days up to 75 days of growth period. Final observations were taken on fresh and dry biomass, plant height (

shoot and root length), leaf number, and mycorrhization in terms of % colonization, spore count and vesicle

number in the treated and the untreated control plants .

Analysis of Growth and AM colonization

Simple biological norms were taken into consideration for the determination of growth parameters including shoo

height, root length, leaf no., leaf area, wet and dry biomass of shoot, root, and leaf, shoot: root ratio.AM infection

and colonization in the roots of maize grown under different treatments were analyzed following the root clearing

& staining technique, and slide method (Phillips and Hayman ,1970; Kormanic and McGraw , 1982). Tota

number vesicles / cm root was also calculated during this experiment. Rhizosphere soil of maize grown unde

different treatments was treated for the AM spore isolation and total number of spore present in 100/g of soi

were counted simply with the help of stereo zoom microscope.

Result

The efficiency of the isolated AM fungi (Glomussp.) was examined along with different watering schedule on

maize plants . Periodical analysis showed progressive plant growth. Daily watering along with inoculation of AM

fungi enhanced shoot height, leaf no. and leaf area at 60 days of sowing ( Fig. 1). Over all, alternate days (W1

watering have given the superior plant growth response while watering at two days intervals (W2) showed

almost similar performance recorded with daily watering schedule along with AM inoculation. At the final stage

of growth measurement, the treatment combination i. e. inoculation with daily watering with AM fungi(C2)

gave the best plant shoot height i. e. 137.78 cm; there was no significant differences between the control (C1

and other treatments ( Fig. 2) . Under the three watering schedule, the plants had almost same number o

leaves / plant . The watering treatment of two day interval (W2) produced longer roots, similarly, fresh and dry

biomass of root was the maximum in this treatment (Fig.3 & 4).

In the present study the untreated and uninoculated plants (C1) which were watered daily did not show any

colonization, the rest of the inoculated plants under different watering schedule showed mycorrhization in


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their roots (Fig.5) . The highest % (93.33) could be observed in the plants watered on alternate days (W1)

followed by daily watering (C2) ( 84 %). However , alternate day watering has given colonization of 33. 33 %.

The total number of vesicles and spores /100 g was also examined which observed to be the maximum in the

plants watered daily followed by plants watered on alternate days or at two day intervals. In the present study

three variable i. e. root length , dry biomass and % colonization were not found to be interrelated . No

significant differences could be observed in the treatments under different watering schedule, but higher S

R length ratio was observed in the alternate watering schedule (W1) ( Fig. 6) . In this study no significantvariation was observed in soil pH .

Data on proportional biomass clearly indicate the progressive enhancement of leaf dry weight under

different watering schedule (Fig.7). Under daily watering schedule total leaf biomass of maize was 15 %

and 27 % in the uninoculated and inoculated controls, where as the leaf yield increase to 44 % and 36 % when

the plants were irrigated at alternate days and/or two day intervals. The maximum shoot biomass ( 42 %) wa

recorded in plants treated with AM fungi under daily watering schedule.

The relative growth rate (RGR) changed in accordance to that of dry biomass, plant height and leaf area

(Fig. 8). It was maximum (0.16 mg -1day 1) in plants of two day alternate watering. Net assimilation rate

(NAR) measured 0.45 mg cm -2 day 1 in uninoculated control (daily watering), 0.43 mg cm -2day 1 in

inoculated (daily watering), 0.56 in alternate days and 0.58 mg cm -2 day 1 two days interval watering

schedule( fig. 9). Leaf area ratio (LAR) is higher in inoculated plants treated with daily watering.

A critical analysis of variance for shoot length at different stages of growth of maize analyzed. The

treatments varied significantly in terms of maize growth periodically (at 0.1% level). Analysis of variance for lea

number of different stages of maize growth under different treatments revealed variation at 0.1 % level. A wide

level of variations (at 1% level) was found through the analysis of variance for leaf area at different stages omaize growth under different treatments

DiscussionHardie and Leyton ( 1981) stated that drought might be relived by an increased rate of root growth and more

efficient extraction of water from soil. Induced stress due to less frequent watering helped in better colonization

(93. 33 %) with more number of vesicles (161.67 ) . Kothari et al. ( 1991) observed that rate of water uptake

per unit root length and per unit tissue by AM were about twice that of non mycorrhizal plant and attributed thi

to hyphal transport. Readhead (1975) reported that the amount of water which was optimal for the growthplants also resulted in the greatest production of fungal spores. Nelsen and Safir ( 1982) reported that the

drought stress reduced spore production. Bethlenfalvay and Franson.(1989) reported that colonization o

mycorrhizal roots didnt vary with stress but the biomass and length of AM was greater in severe stress .

Results revealed that AM inoculation improved drought tolerance of maize. This findings is in

agreement with the findings of Subramanian and Charest ( 1997) who reported that AM colonization enhanced

N and P uptake that resulted in higher shoot biomass production under drought condition .Watering on alternat


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days and at two day intervals helped in production of more leafy biomass by 21. 4 % and 36 % , respectively

which was comparable to the daily watering schedule under mycorrhization conditions; the data corroborated

the findings of Subramanian et al. ( 1995) who reported that AM inoculation enhanced maize growth and

enabled the host plants to sustain moderate drought conditions.

Improvement in the crop production due to introduced AM isolates was coupled with improvement in

the mycorrhization status. However, in certain cases isolates improved the host performance but failed toimprove the mycelial status. At the same time, in certain other cases an isolate failed to improve the

performance of the crop in spite of its ability to raise the mycorrhizal status. It has been stated that mycorrhiza

might impart better plant growth under dry as compared to the wet conditions (Michelsen and Rosendahl

1990). The present results confirmed the above findings on maize.

Plants inoculated with AM fungi produced more dry biomass yield than the nonmycorrhizal plants. The increase

in plant growth in mycorrhiza treated plants was probably due to enhancement of P uptake ( (Karaki and

Raddad, 1997); under water stress condition, mycorrhizal plant also gave more biomass. This results arecontrary to reports of Karaki and Raddad (1997) who reported that low biomass yield of wheat due to AM

inoculation under drought condition. It was suggested that mycorrhiza were more useful for plant growth unde

dry conditions than the wet conditions ( Michelsen and Rosendahl, 1990).

Acknowledgements : Authors are thankful to Department of Forests and Environment , Govt. of Orissa for

various helps.

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Effect of Water Stress on Performance of Maize Inoculated With Glomus Sp

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