Jan. 05, 2007Dr. Intisar E. Elteraifi1 Intisar E. Elteraifi, (Ph.D.) Assistant Professor, Forest...

Jan. 05, 2007 Dr. Intisar E. Elteraifi 1

Intisar E. Elteraifi, (Ph.D.)Assistant Professor, Forest Ecology

Environmental Sciences & natural Resources Department

Faculty of Agricultural SciencesUNIVERSITY OF GEZIRA

P.O Box 20, Medani, SUDAN

E-mail:[email protected]


Research projects

Research at GU/FAS/ESNRD

Other institutes of similar interest in Sudan

Running research projects


Geographic coordinates22 longitudes 22 to 38 East and latitudes 4 to 22 North

Population: 41,236,378 (July 2006 est.)

Area:Total: 2,505,810 sq km Water: 129,810 sq km Land: 2.376 million sq km


Natural resources:petroleum; small reserves of iron ore, copper, chromium ore, zinc, tungsten, mica, silver, gold, hydropower Land use:arable land: 6.78% permanent crops: 0.17% other: 93.05% (2005) Irrigated land:18,630 sq km (2003) Natural hazards:dust storms and periodic persistent droughtsEnvironment current issues:inadequate supplies of potable water; wildlife populations threatened by excessive hunting; soil erosion; desertification; periodic drought Environment - international agreements:party to: Biodiversity, Climate Change, Climate Change-Kyoto Protocol, Desertification, Endangered Species, Law of the Sea, Ozone Layer Protection signed, but not ratified: none of the selected agreements Geography - note:largest country in Africa; dominated by the Nile and its tributaries


Sudan has turned around a struggling economy with sound economic policies and infrastructure investments, but it still faces formidable economic problems, starting from its low level of per capita output. From 1997 to date, Sudan has been implementing IMF macroeconomic reforms. In 1999, Sudan began exporting crude oil and in the last quarter of 1999 recorded its first trade surplus, which, along with monetary policy, has stabilized the exchange rate. Increased oil production, revived light industry, and expanded export processing zones helped sustain GDP growth at 8.6% in 2004. Agricultural production remains Sudan's most important sector, employing 80% of the work force, contributing 39% of GDP, and accounting for most of GDP growth, but most farms remain rain-fed and susceptible to drought. Chronic instability - resulting from the long-standing civil war IN south, adverse weather, and weak world agricultural prices - ensure that much of the population will remain at or below the poverty line for years


Population below poverty line:40% (2004 est.) Unemployment rate:18.7% (2002 est.) GDP - composition by sector:agriculture: 38.7% industry: 20.3% services: 41% (2003 est.) Labor force:7.415 million (1996 est.) Labor force - by occupation:agriculture: 80% industry: 7% services: 13% (1998 est(


Major infectious diseases:degree of risk: very high food or waterborne diseases: bacterial and protozoal diarrhea, hepatitis A, and typhoid fever vectorborne diseases: malaria, dengue fever, African trypanosomiasis (sleeping sickness) are high risks in some locations water contact disease: schistosomiasis respiratory disease: meningococcal meningitis (2005)


• Deforestation

• Decrease in Biodiversity (Flora/Funa)

•Urbanizations

•Drought

•Soil Degradation

•Pesticides (Pollution)

•Energy Crises

•Refugees & Displacement (War in South/Darfur)

•Floods/as a seasonal problem (The Nile F.P. on Sept.)


History of Science and Technology in Sudan

Since the independence (1956), science and technology policies were not formulated in a comprehensive sense, but left to the institutional and academic initiatives. The first organized structure of science and technology came out with the establishment of National Council for Research in 1970. The National Council for Research is the body that was responsible for the formulation of policies and application of research for the societal benefit and development. Until 2001 efforts were made to initiate technology capabilities in various organizations in the country, however, the outcomes were not satisfactory due to many constrains.


Sudan is a vast country spans over 18 degrees of latitude from the desert in the north with hot dry climate to the sub-tropical region in the south with heavy rains and dense vegetation cover. This endows the country with diversity as reflected in various environments and natural resources. The country enjoys extensive arable land that makes agriculture the leading sector and the backbone of industry. The natural resources have also allowed the build-up of a natural herd of livestock as well as wild life. Beside, the country is well known for its rich mineral resources including gold, iron lead chrome…etc, and very recently oil production supports the economy of the country.


Universities In SudanKhartoum University:

The establishment of a campus for Cairo University in Khartoum followed it and in the early sixties, Khartoum Polytechnic Institute was founded. Gezira University is the 3rd .

In recent years, higher education has greatly grown with the government establishing a number of universities in Khartoum and in other states of Sudan. The number of Sudanese Universities now are about 26.

Private Universities:

The private sector is notably contributing in the advancement of higher education in Sudan.

Ahfad University for Women, founded by the well-known advocate of women's education, Sheikh Babikir Badri, is considered to be the pioneer in this field.

Private and popular higher education boasts studies of modern technology, languages, accountancy, statistics and other disciplines.

There is 26 state university and main 6 universities beside large number of private colleges in different areas of science and technology.


Universities In Sudan


Post graduate studies

M.Sc & Ph.D.’s programs are running in most of these universities and research institutes beside the undergraduate programs.

International fellowships & local funds are also available e.g.. DAAD, TWAS, TWOWS, CIRAD, ICRAF, DANNIDA, FINNIDA.


Chemical Ecology studies in Neem, Striga & sorghum, Mesquit….etc

Screening plants for anti-malarial products.

Forest Conservation (Forest Ecology In Acacia Senegal)

Mapping of the NR in Gezira State

(Desert & Karab encroachment)


Agro forestry for improving the local microclimate.

Agricultural Residues as a source for energy sources.

Soil Seed Banks: Evaluation for natural regeneration.

Evaluation of different methods used in desert compaction.


TTMI (Netherlands) Agro-foretry (ICRAF-Kenya) Natural Resources management (France) Water resources management at field level Bio-pesticides (screening) Soil Seed bank (Joint project with FNC-FAO) Medicinal Plants Renewable energy resources Drought resistant crops Alellopathy (plants-animals interactions)


Future Collaboration Between University of Gezira, Sudan & IISC Chemical Ecology Forest Ecology Environmental Conservation Natural Resources management Marin Ecology Drought resistance Natural products

VARIABILITY IN GROWTH, OIL AND VARIABILITY IN GROWTH, OIL AND LIMONOIDS IN NEEM (AZADIRACHTA LIMONOIDS IN NEEM (AZADIRACHTA INDICA A. JUSS) FROM DIFFERENT INDICA A. JUSS) FROM DIFFERENT

ECOZONES IN SUDANECOZONES IN SUDAN


The increasing cost of the synthetic PESTICIDES & the growing awareness of hazards associated with their large-scale use have evoked a worldwide interest in pest control agents of

PLANT ORIGIN

BACKGROUND


NEEM TREE

The Village pharmacy

The Nature’s gift to mankind

The wonder tree

A tree for solving global problems

The Global Tree


Neem tree is an important exotic tree in Sudan which

contributes in forest sector as

shade, firewood, pesticide & medicinal

plant.


NEEM & AGROFORESTRYNEEM & AGROFORESTRY

Neem has been selected to be among the best trees for Agro-forestry systems in dry & semi dry zones.


Neem is highly suitable for the improvement of degraded and nutrient-poor soil.

Important in reforestation programs to combat desertification.

Leaves mulch improves the soil-water conservation, reduces the soil temperature & releases nutrients through decomposition.

Neem cake increases the productivity & works as nematocides


GLOBAL INTEREST ON NEEMGLOBAL INTEREST ON NEEM

There have been seven International Neem There have been seven International Neem

Conferences. All concluded thatConferences. All concluded that : :

Neem is the best promising plant for Neem is the best promising plant for

bio-pesticides productionbio-pesticides production..


Extracts of neem fruit, seeds, seed kernels, twigs, stem bark & root bark have:

Insecticidal [insect antifeedant, insect growth disrupting]

Nematocides Fungicidal Bactericidal Anti-inflammatory Anti-tumor Immune-stimulating Anti-malarial


What is in NeemWhat is in Neem???? Since the early investigation of

Siddiqui (1942), more than 300

compounds have been isolated from

various parts of the neem tree .

Azadirachtin (is the most

biologically active compound), Salannin,

Nimbin & Gedunin


O

OO

O

H

HO

OO

O

O

O

O

H

OHOMe

O O

OOH

O

O

O

O O O

OAc

(1)

(4)(3)

O

O

O

O

O

(2)

O

O

O

O

AcO

OHOMe

O O

OOH

(5)

Scheme 1 (1) Nomilin. (2) Citrolin. (3)Harissonin. (4) limonin. (5) Acetoxylharrisonin.


O

O

OH

OH

AcO

OO COOMe

OH

HO

OH

O

MeOOC

O

MeO2C

OR'

R''O

O

R' = Tig R'' = Ac

R' = Tig R'' = H

O

O

MeO2C

O

OAc

HO

O

HO

OH

HOH

O OR

O

OO

O

R=Ac

R=H

(6)

(8)

(9) (10)

(12)

(11)

(7)

COOMe

Scheme 2 (6) Azadirachtin. (7) Nimbin. (8) Salannin. (9) 3-deacetylsalannin. (10) Meliantriol. (11). Gedunin. (12) 7-deacetylegedunin.


Azadirachtin It was isolated by Morgan in 1968

from neem seeds, who showed that it

has the formula C35H44O16.

This compound is now known to affect

over 200 species of insects and mites.

Many isomers of azadirachtin, Az-A to

Az-H are isolated till now.

AcOH

O

OH

O

O

CH3OOCTigO

OH

O

OH

OH

CH3OOC

Azadirachtin


Neem Products


The exact origin of A. indica is not known. Many authors suggest that it may lie

in Myanmar (Burma) &/or in southern parts of India.

Nowadays, neem is widely distributed by introduction mainly in tropical &

subtropical zones of Asia, Africa, The Americas, Australia & the South

Pacific Islands.

Origin & Distribution

Jan. 05, 2007 Dr. Intisar E. Elteraifi 33Distribution of the neem tree at the source of origin


EcologyEcologyNeem tree: Thrives in areas with arid, sub-arid to sub

humid conditions .

Rainfall : 400-1200 mm (less than 400 mm Depends on the ground water, 130 mm. up to 2500 depends on soil type.) Neem failed to grow in Tonga Island in

Ecuador/Amazon basin (3000-4000 mm) .

Soil : It grows in wide variety of neutral to alkaline soils

Well drained, shallow, stony, sandy soils. In Sudan & India it thrives on black cotton soils .

Altitude 0-1500 m


Pesticides hazards

Agricultural pests

Needs for More food

High cost

Sudan is an agricultural country


OBJECTIVESOBJECTIVESOverall Objective

The main objective of this study is to generate requisite information, which would provide the basis of exploiting the neem tree in Sudan as a source of safe, ‘soft’ pesticides, as well as other raw materials.


Specific objectives

To study the characteristics of the neem seeds collected from ten seed zones in Sudan.

To investigate the establishment and growth of ten neem ecotypes from ten seed zones in Sudan under nursery conditions.


To study the effect of two irrigation regimes on ten neem ecotypes from different seed zones in Sudan under nursery conditions.

To quantify and evaluate the total oil content in neem seed kernel collected from ten seed zones in Sudan.

To quantify three active ingredients (limonoids) in NSK methanol extracts specifically azadirachtin (A and B), salannin and nimbin from 10 zones in Sudan over two seasons.

To establish relationship between limonoids and the climatic factors &/or GPS, if any, to study the agro-ecological attributes.

To produce predicted quality maps for Az, Sa & Ni in Sudan.


ExperimentsExperimentsTen ecological zones were selected according to the tree seed zoning systems map (DANIDA/ARC, 1995).

Four sites within each zone were selected, (15-20) trees were marked randomly in each site. (2-3) kg of the neem fruits were collected from these trees.

Jan. 05, 2007 Dr. Intisar E. Elteraifi 42The selected seed zones in Sudan


Climatological data & GPS

RF, Temp. , RH% and GPS data for the selected agro-ecological zones were obtained from the meteorological Department, Sudan.


Ecological zones for neem seed collections Seed Zone RF (mm) Temp. (oC) RH%

S1 12.3 27.3 25

S2 39.4 29.4 25

S3 162.4 29.9 29

S4 306.4 28.6 39

S5 318.0 27.3 34

S7 581.1 28.8 45

S8 712.9 28.3 47

S9 398.3 27.2 31

S10 633.1 28.1 47

•Data from the Meteorology Department, Sudan. •Climatological normals (1961-1990)


Annual rainfall and the selected zones in Sudan

6

12

8

43

9

75

10


1 .Seed measurements1 .Number of seeds per kilogram

2 .Seed size (length & width)3 .Seed, seed kernel & shell weights.

2 .Nursery Experiments 1 .Provenance variation

2 .Drought tolerance studies3 .Quantification of N.O. in NSK

4 .Limonoids 1 .Azadirachtin

2 .Salannin 3 .Nimbin


Seed MeasurementsNumber of seeds per kilogram

Seed dimensions (Seed length & width)

Seed, seed kernel & shell weights

for each ecotypes were carried out according to ISTA (1996) .


Seedling Raisings

Neem seeds were sown in soil medium (2 clay: 1 sand) in polythene pots (20x25 cm and 15 mm thickness) placed under shade (traditional nursery) at 3-5 seeds per pot. Conducted at the NTSC/ ARC located at Soba/ Sudan.


Experimental design (4x3x8x20).Four blocks, 3 replicates. 8 provenances; represented by 20 seedlings in a Randomized Complete Block Design (RCBD).

Provenance variation Sowing date was 25/7/1999; seedlings were irrigated every day for 12 weeks by using flood irrigation.

Measurements

Destructive measurements were conducted every four weeks after establishment of 12 weeks. Data of 7 ages ( A1-A7) were collected after removal of the seedlings by using clean water, using mean of three seedlings in each measurement.


Morphometrics characters of seed and seedlings measured.

Seedling Leaf Seed -seedling height (SH) -total no. of leaves per

seedling (TL) -seed length (SL)

-collar diameter (CD) -leaf length (LL) -seed diameter (SD)

-root length (RL) -leaflet ratio (length: widths) (L:W)

-seed weight (SW)

-no. of roots (R No.) -no. of seed per Kg

-distance between cotyledons and the1st two

leaves (DCL)

-kernel weight

-shoot dry weight (SDW)

-root dry weight (RDW)

-shoot: root ratio (S: R)


IRRIGATION

Ten healthy seedlings of uniform height were chosen from each

provenance .

2 blocks; 3 replicates, 8 provenances, 10 seedlings, in randomized complete block

design (RCBD) (2x3x8x10).


Seedlings in block I were watered every 10 days, while the seedlings in block II were watered every day.

Block I Block II


The seedlings were harvested after 27 weeks under the irrigation regimes.

the following measurements were made:

Shoot height Root length Shoot & root dry weight Survival percentage


DATA ANLYSES Data of seed morphometerics, provenance

evaluation and irrigation regimes experiments at the nursery were subjected to analysis of variance (ANOVA) &principal component analysis (PCA) using SAS software (SAS Institute Inc., V8, North

Carolina, USA) .

The multiple comparison procedure using Turkey’s Least Significant Difference (LSD) test (P=0.05) (SAS, ver. 8, 1998) was used to compare the provenance means.


Oil ExtractionExtraction was carried out using

Soxhlet apparatus. 3 g of NSKP were placed in a thimble and 500 ml of hexane were used to extract the oil. Each sample was replicated twice.

Neem oil percentage was calculated by weighing the flask before and after extraction.


Soxhlet apparatus

wt1= weight of the neem kernel powderwt2= weight of the empty flaskwt3 = weight of the flask with oil

1001

23X

wt

wtwt Neem oil% =

The flask


LIMONOIDS EXTRACTION

Plant materialCleaned & dried NS were taken to BCED

laboratories at ICIPE (Nairobi-Kenya), where they were crushed with a pestle in a mortar to

obtain the kernels .seed kernels were ground in an electric blender

to prepare the neem seed kernel powder (NSKP) which was used in the analyses.


1

5 4

32

Chemicals Laboratory-grade reagents & solvents were

used in extractions; methanol (CH3OH) purity 99%, hexane (CH3 (CH2)4CH3) (purity

99.8%) and DD water .

Extraction was done in two replicates; 12 gm of each sample was extracted with 25 ml of methanol in a 250 ml conical flask and stirred magnetically for 2 hrs at room temp (cold extraction). The extraction was repeated two times (for 1 hr each).


The pooled methanol extract was shaken with 30 ml of hexane and 1.0 ml of DD

H2O .Extraction with hexane was repeated

two times .Two layers were obtained in each case,

the upper (mainly hexane) layer was separated from the lower aqueous methanol layer which was concentrated in vacuo to 10 ml and stored at (-15C) for HPLC analysis.

Schematic diagram of Rotavapor (Büchi 461, water bath, Switzerland)


CHROMOTOGRAPHY

Beckman HPLC, (System Gold): ODS columns, ultrasphere C-18, 2504.6 mm

Prog. 40% acetonitrile in water (10 min.) to 70% acetonitrile (10 min.) & finally to 100%

acetonitrile (5 min.), all at 1 ml/min .The eluents were monitored at 214 nm .20 µl of the sample were injected.Each sample was analyzed three times.Quantification of the limonoids was based on

injections of known quantities of standard samples.10 zones X (4 Sites) X (2 samples) X (3

Injections)=240


Analytical High Performance Liquid Chromatography (Beckman HPLC, System Gold).


The standard substances of azadirachtin (A and B), salannin & nimbin were purchased from Trifolo-M-GmbH (Germany).

1.00 µg of each was dissolved in 5 ml of methanol.

Source and preparation of the

Standards


E_Zone Site Sample Inj_No PA_AZB PA_AZA PA_NI PA_SA AZ_ABdong 1 1 1 11.21045 296.5744 491.7935 575.9929 307.7849dong 1 1 2 13.11693 113.2527 415.9136 477.8567 126.3696dong 1 1 3 14.00935 84.88821 420.7505 482.9827 98.89756dong 1 2 1 16.40044 106.2245 712.5898 794.0378 122.625dong 1 2 2 20.24448 107.688 733.5112 811.8212 127.9324dong 1 2 3 352.2315 247.9885 693.698 771.9911 600.22dong 2 1 1 341.2219 160.7103 739.1872 844.3686 501.9322dong 2 1 2 326.6384 217.0517 774.9457 863.769 543.6901dong 2 1 3 315.1511 206.2006 766.9013 843.9926 521.3517dong 2 2 1 336.2609 211.6228 787.0356 873.8317 547.8837dong 2 2 2 285.6296 203.5535 737.0663 815.6075 489.1831dong 2 2 3 302.1174 207.1877 775.4281 850.2117 509.3051dong 3 1 1 294.1361 208.3945 737.6437 826.2441 502.5307dong 3 1 2 291.995 213.0284 735.0895 804.4811 505.0234dong 3 1 3 292.0742 219.1079 744.6856 809.5714 511.1821dong 3 2 1 343.6914 261.8198 607.9752 791.3439 605.5113dong 3 2 2 335.5088 170.2895 715.3398 794.1877 505.7983dong 3 2 3 420.7048 197.2497 1431.377 861.3019 617.9545dong 4 1 1 506.5932 240.7411 795.8378 935.3828 747.3342dong 4 1 2 128.2045 44.6694 831.876 975.3163 172.8739dong 4 1 3 126.0951 50.20505 838.847 980.0244 176.3001dong 4 2 1 113.8354 49.05298 820.1082 962.8843 162.8883dong 4 2 2 115.3013 46.36087 807.3794 940.1558 161.6621dong 4 2 3 729.5483 4.74212 798.3073 931.2002 734.2905


CALCULATION OF THE LIMNOIDS LEVEL IN NSK

MCW

DfVDS

100

100

xS = limonoid content of the sample [mg/g dry matter]xD = limonoid content in the dilution [mg/ml]V = total volume of the extract [ml]Df = dilution factorW = sample fresh weight [g]MC = mean weight loss on drying [%]

According to the formula:


High performance liquid chromatography of the neem seed kernel methanol extract


Data of the three compounds (Az , Sa & Ni) and the total were subjected to analysis of variance (ANOVA) & principal component analysis (PCA) using SAS software (SAS Institute Inc., V8, North Carolina, USA). Tukey’s Honest Significant Difference (HSD) test was used to compare the limonoid content in the different zones.

DATA ANALYSES


Correlation and multiple regression analysis were

also carried out using SAS to find the possible

statistical relations between the meteorological data,

GPS and the limonoid contents. GLM and PCA

procedures were also used .

Spatial analysis methods: Inverse distance weight:

nearest neighborhood: power. Were used for

production of the GIS predicted maps for the

limonoids.

DATA ANALYSES


KEY RESULTSKEY RESULTS


Significant variations (p<0.0001) were found between

the ten zones in the seed parameters: no. of seed/kg,

seed length, seed width, seed & kernel weights

No significant differences were found between sites

within the zones & between trees within the same

site.

Seed kernel dry weight was found to be varying from

97.26 mg to 38.22 mg; while that of the seed shell

varied from 128.5 mg to 75.29 mg.

Seed parametersSeed parameters


0

2000

4000

6000

1 2 3 4 5 6 7 8 9 10

Seed zones

Nu

mb

er

of

see

ds/

Kg

.Season 1

Season 2

Variations in number of neem seeds/Kg. collected from various seed zones in Sudan over two seasons 2000-2001 and 2001-2002


Seed variations between trees within the same site in the Sudan

Seed Length (cm) Seed width (cm) Number of seeds/kg

Tree no. )Mean± SE( )Mean± SE( )Mean± SE(

1 0.630 ±0.005a 1.199 ±0.041a 5832.510 ±170.191a

2 0.642 ±0.005a 1.187 ±0.009a 5444.200 ±33.531a

3 0.614 ±0.039a 1.192 ±0.025a 5621.790 ±50.589a

4 0.617 ±0.004a 1.174 ±0.007a 5742.070 ±64.222a

5 0.629 ±0.003a 1.187 ±0.005a 5675.560 ±83.824a

6 0.628 ±0.006a 1.186 ±0.011a 5584.570 ±80.435a

Means followed by the same letter in the same column are not significantly different at P<0.05 (Tukey’s HSD test).


Variations in neem seed length and seed width in 10 seed zones in Sudan

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1 2 3 4 5 6 7 8 9 10

Seed zones

Se

ed

le

ng

th &

wid

th i

n c

m

S. Length S. width


K.WT SH.WT

Contribution of the seed shell and seed kernel in the neem seed weight


0

20

40

60

80

100

120

140

S1 S2 S3 S4 S5 S6 S7 S8 S9 S10

seeds zone

We

igh

t (m

g)

kernel w t

shell w t

Variations on neem seed kernel and shell weight in different zones in Sudan


No significant variations were found between the

ten provenances in most of the morphometrics

parameters measured.

Variations between ages of the ten provenances

were significantly different but no variations were

found within the same age.

Slight variations were observed in leaves and root

length .

Provenance VariationsProvenance Variations


Morphological variations in some neem provenances in Sudan. a) Seeds ; b) leaves ; c) roots.

a b

c


0

0.2

0.4

0.6

0.8

1

1.2

DCL cm

S1 S2 S3 S5 S6 S8 S9 S10

Seed sources

DCL

Variations in the distance from the first two leaves and the cotyledons scars (DCL) in neem seedlings from different seed zones in Sudan


Results showed highly significant

differences (P<0.0001) between the

two irrigation regimes in almost all

parameters measured; seedling

height, root length, shoot dry-

weight, root dry-weight and survival

percentage over the two seasons.

Irrigation RegimesIrrigation Regimes


No variations were found between

the seed sources within the same

treatment (irrigation regime) in the

measured traits.

There was a clear morphological

difference between the two regimes of

irrigation in terms of growth and the

colour of the leaves


0

10

20

30

40

50

60

70

S1 S2 S3 S5 S6 S8 S9 S10

Seed sources

Sh

ho

t h

eig

ht

Irrigation I Irrigation II

Effects of two irrigation regimes on shoot height of neem seedlings from various ecozones.


0

10

20

30

40

S1 S2 S3 S5 S6 S8 S9 S10

Seed sources

Ro

ot

len

gth

Irrigation I Irrigation II

Effects of two irrigation regimes on root length of neem seedlings from various ecozones.


0123456789

10

S1 S2 S3 S5 S6 S8 S9 S10

Seed sources

Sh

oo

t d

ry w

eig

ht

(g)

Irrigation I

Irrigation II

Effects of two irrigation regimes on shoot dry weight of neem seedlings from different ecozones.


Effects of two irrigation regimes on root dry weight of neem seedlings from different ecozones.

0

1

2

3

4

5

6

S1 S2 S3 S5 S6 S8 S9 S10

Seed sources

Ro

ot

dry

we

igh

t (g

)

Irrigation I

Irrigation II


Effects of two irrigation regimes on neem seedlings from various ecozones (a) Irrigation I & (b) Irrigation II

a) b)


Morphological variations between the two irrigation regimes in the root system of two seed sources: (a) S9 & (b) S3.

a) b)


Variations between two irrigation regimes on survival % of various neem ecotypes in Sudan

0

20

40

60

80

100

120

S1 S2 S3 S5 S6 S8 S9 S10

Seed sources

Su

rviv

al

%

Irrigation I

Irrigation II


Effects of two irrigation regimes on the seedling height, root length, shoot dry weight, root dry weight and survival percentage (season 2001-2002).

Regime Sh. H)cm(

R.L)cm(

Sh. D. Wt.)g(

R. D. Wt.)g(

Survival%

I 42.03 b 24.32 b 3.37 b 2.25 b 63.33 b

II 50.24 a 30.56 a 5.95 a 3.97 a 94.17 a

Regime I = Irrigation every 10 days.Regime II = Irrigation every day

Means followed by the same letter in the same column are not significantly different (P<0.05). (Tukey’s HSD test).


Effects of two irrigation regimes on the neem seedling parameters (season 2002-2003)

Regime Sh. H)cm(

R.L)cm(

Sh. D. Wt.)g(

R. D. Wt.)g(

Survival%

I 36.50 a 26.58 b 3.16 b 1.39 b 65.53 b

II 34.02 a 31.35 a 4.47 a 1.75 a 90.14 a

Regime I = Irrigation every 10 days Regime II = Irrigation every day

Means followed by the same letter in the same column are not significantly different (P<0.05). (Tukey’s HSD test).


Overall effect of the two types of irrigation regimes on neem ecotypes in Sudan.

Zones Sh. H R. L Sh.D wt R.Dwt Survival

S1 48.023)3.880 a(

28.117)3.600 a(

5.786)1.859a(

3.569)1.479 ab(

90.000)1.309 a(

S2 44.813)3.813 a(

26.100)3.296 a(

4.376)1.648 ab(

2.852)1.335 ab(

76.670)0.949 a(

S3 50.133)3.933 a(

26.883)3.311 a(

4.733)1.727 ab(

3.222)1.430 ab(

83.333)1.223 a(

S5 45.067)3.813 a(

27.883)3.358 a(

4.685)1.712 ab(

2.918)1.338 ab(

93.860)1.356 a(

S6 49.450)3.907 a(

28.117)3.358 a(

5.507)1.824 ab(

3.854)1.555 a(

86.667)1.202 a(

S8 40.350)3.671 a(

26.100)3.305 a(

3.096)1.310b(

2.674)1.236b(

6.000)0.738 a(

S9 41.683)3.706 a(

26.017)3.287 a(

4.120)1.515 ab(

2.439)1.197b(

66.667)0.900 a(

S10 49.483)3.903 a(

29.783)3.407 a(

4.978)1.732 ab(

3.350)1.422 ab(

73.333)1.034 a(

Means followed by the same letter in the same column are not significantly different at the 5% (P<0.05). (Tukey’s HSD test).. Adjusted mean test on transformed mean values. Values of the transformed data are in the prances


N.O. percentages are found to vary from

41.1±2.9 % to 47.4±2.1 % (at 5% mc) .

The average yield of the country was

found to be 44.6%.

No variation was found between the

different regions in the oil-contents.

Neem oil (N.O.)Neem oil (N.O.)


The limonoids results presented in

this section are AZ (the sum. of AZ-

A + AZ-B), salannin, and nimbin.

The total amount of the limonoids

was the summation of the three

limonoids.

LimonoidsLimonoids


Significant (p<0.0001) variations were found in

AZ- level between the different zones .

AZ-content varied from 1.08 mg/g to 2.34

mg/g in 2001 and from 0.48 mg/g to 3.091

mg/g in 2002 .

Significant variations between the different sites

within the zones, and between the trees within

the sites were found.

Variation in the AZ- level Variation in the AZ- level between zonesbetween zones

Jan. 05, 2007 Dr. Intisar E. Elteraifi 95Variation in the azadirachtin level (mg/g) in seeds from different zones in Sudan for two seasons 2001 and 2002.

Columns with the same letter are not significantly different at p≤0.05


Variations in the total amount of limonoids and AZ in the extract of NSK from ten zones in Sudan.

Zone AZ (mg/g)* Total (mg/g)* S1 1.0771 0.0736e 4.1286 0.2154c

S2 1.2454 0.05286de 4.4631 0.0848c

S3 1.8696 0.08677bc 6.2766 0.213b

S4 1.4884 0.0906cde 4.8287 0.26c

S5 1.5064 0.097cd 4.8066 0.2462c

S6 2.0969 0.0989ab 6.6798 0.1276ab

S7 2.0995 0.1649ab 6.6354 0.2754ab

S8 2.2426 0.1544ab 6.9247 0.2847ab

S9 2.3444 0.1531a 7.1594 0.2922a

S10 1.8184 0.0963bc 6.2897 0.1636b

Means followed by the same letters in the same column are not significantly different (p<0.05), Tukey's Student Range (HSD) test. .* mean± SE


Significant variations were found

between different zones in both

salannin & nimbin (p<0.0001) over the

two seasons.

Variations were also observed between

sites within the zones and between trees

in the same site.

Variation in Salannin & Variation in Salannin & Nimbin levels between zonesNimbin levels between zones

Jan. 05, 2007 Dr. Intisar E. Elteraifi 98Variations in salannin level in different zones in Sudan over two seasons


Variations in nimbin level in different zones in Sudan over two seasons


Simple and multiple regressions were

used to find out the possible relations of

meteorological factors with AZ-level

(concentration) in NSK collected from

10 different zones .

Effect of the Meteorological & GPS Effect of the Meteorological & GPS data on limonoids Level in NSKdata on limonoids Level in NSK


The rainfall was significantly affecting

AZ-concentration in NSK.

Altitude showed a significant positive

effect on AZ-content.

No significant temperature effect on the

level of the AZ in NSK was observed.

The R.H. & temperature showed no

significant correlation with AZ-level.


Pearson correlation coefficients of the ecological factors and the limonoids concentrations.

AZ Sa Ni Total

Temp -0.006NS 0.095 NS 0.099 NS 0.002 NS

)0.880( )0.622( )0.816( )0.997(

RF 0.738* 0.786* 0.378 NS 0.726*

)0.037( )0.021( )0.356( )0.042(

R H 0.513 NS 0.616 NS 0.115 NS 0.511 NS

)0.194( )0.102( )0.787( )0.196(

Alt. 0.725* 0.551 NS 0.561 NS 0.644 NS

)0.042( )0.157( )0.148( )0.085(

Lat. 0.790* 0.765* -0.487 NS -0.756*

)0.020( )0.027( )0.221( )0.030(

Values of P are shown between the brackets. ; NS: Not significant ; * Significant (P< 0.05)

Jan. 05, 2007 Dr. Intisar E. Elteraifi 103The trend of azadirachtin level in neem seed kernels with the rainfall in Sudan

The optimal rainfall = 717 mm


Rainfall was found to have significant

(P=0.02) and positive correlation with

salannin and total amount of limonoids in the

NSK.

Temperature, relative humidity and altitude

were found to have no significant correlation

with both of the salannin and nimbin.

Salannin content was found to be significantly

(P=0.02) affected by cross products of

temperature and rainfall.

Jan. 05, 2007 Dr. Intisar E. Elteraifi 105Relationship between the rainfall and total amount of limonoids.


Relationship between the rainfall and Salannin.

Jan. 05, 2007 Dr. Intisar E. Elteraifi 107The combined effect of rainfall and temperature on the level of salannin.

Sa=1.74513+6194*10 -4 *(RF*Temp) (P=0.02)


Three maps were produced

for the predicted levels of AZ,

salannin and nimbin in Sudan.

These maps were produced

using spatial analysis methods.

Mapping and GIS

Jan. 05, 2007 Dr. Intisar E. Elteraifi 109Az = - f (latitude), Except Khartoum

Incr

ease

in A

z

Predicted concentration of azadirachtin in Sudan as determined by GIS.

0 500 Km

Jan. 05, 2007 Dr. Intisar E. Elteraifi 110Sa = - f (latitude), except Khartoum

Incr

ease

in S

a

Predicted concentration of salannin in Sudan as determined by GIS.

0 500 Km

Predicted concentration of nimbin in Sudan as determined by GIS.

Ni = - f (longitude)

Increase Ni

0 500 Km


The present study showed that morphological

variations occur between neem seeds from

different provenances, but no significant

morphological variations were found in the

corresponding seedlings at the nursery level.

Nursery neem seedlings from different ecozones

did not show any discernible variations in

response to two irrigation regimes.

ConclusionsConclusions


No variations were found in oil

content in neem kernel of seeds from

different ecozones.

The amounts of azadirachtin,

salannin, nimbin and the total amount

of limonoids in the neem seed kernels

from different ecozones were

significantly different.


Azadirachtin was found to be affected by

rainfall, increasing with rainfall in a hyperbolic

fashion reaching an optimum level at ~717 mm.

Salannin levels were correlated with rainfall

and showed greater variation between different

ecozones.

Nimbin showed similar variations between

ecozones, but this did not correlate with

climatic factors (the reasons for which are

unclear).


Previous screening of seed kernels from

trees located in different continents

(drawn from a larger genetic pool) gave

limonoid levels that could not be

correlated with climatic factors. The

present study, which is based on a

presumably narrow pool, shows that

climatic factors are an important set of

determinants for the level of limonoids

found in neem trees.

ielteraifi

Ermel, 1995


This study showed variations between ecotypes

with respect to the contents of Az & salannin in

NSK in relation to climatic factors. For

exploitation of limonoids for bio-pesticides

production, the best source of seeds are from

neem trees growing in humid and semi-

humid zones where the limonoids

content are expected to be high.

RecommendationsRecommendations


Neem seedlings in the nurseries proved to

be quite resistant during drought periods;

therefore, it is recommended that

minimum water inputs may be used at this

stage.

Looking at more samples from other

ecozones in Sudan (e.g. southern parts) is

recommended to extend the screening

programme to other climatic zones in the

country.


Whether these variations result principally

from environmental factors or represent

interactions between genomic differences and

climatic factors need to be resolved by two

kinds of studies: screening genetic differences, if

any, between neem provenances, and long-term

studies of the performance and phytochemical

attributes of plants derived from seeds from one

provenance growing in other provenances .

RecommendationsRecommendations


Detailed studies on other

potentially useful limonoids from

neem leaves and bark (e.g. anti-

malarial constituents such as

gedunin) would also help in

deciding the best sources of these

phytochemicals.


Gedunin Quantification (from neem leaves and stem bark collected from the 10 seed zones in Sudan).

Genomic DNA study for neem leaves samples from the selected zones using RAPID-PCR

Running Research work (Not complete yet)


Future plans

Chemical Ecology : India is rich country in plant biodiversity looking for natural chemicals for disease and insect management could be of interest to do more research work with collaboration of the Indian scientist at the IISC as one of the best center of excellence in south.


Future plans

Desertification and Karab encroachment is one of the serious issues in Karnataka I observed from my field trip with Energy Group. More serious situation in Sudan especially in Gezira State, where is the most important irrigated scheme in Sudan. Collaborative research on this could be of great interest.


Future plans

Survey for plants using the local knowledge & believes will be the 1st step in collection.

screening in laboratory and extraction will be followed by in vivo bioassay.

Using Arial photos and satellite images for the last 50 years in comparison by the recent maps will be the methodology for the predication of the changes could be running in the two states (Gezira, Sudan compare to Karnataka, India)


Future plansComing

back to India Karnataka

is interesting

for me

Jan. 05, 2007Dr. Intisar E. Elteraifi1 Intisar E. Elteraifi, (Ph.D.) Assistant Professor, Forest...

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