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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]
Jan. 05, 2007 Dr. Intisar E. Elteraifi 2
Research projects
Research at GU/FAS/ESNRD
Other institutes of similar interest in Sudan
Running research projects
Jan. 05, 2007 Dr. Intisar E. Elteraifi 3
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 4
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 5
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 6
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(
Jan. 05, 2007 Dr. Intisar E. Elteraifi 7
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 8
• 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.)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 9
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 10
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 11
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 12
Universities In Sudan
Jan. 05, 2007 Dr. Intisar E. Elteraifi 13
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 14
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 15
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 16
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 17
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 19
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 20
NEEM TREE
The Village pharmacy
The Nature’s gift to mankind
The wonder tree
A tree for solving global problems
The Global Tree
Jan. 05, 2007 Dr. Intisar E. Elteraifi 21
Neem tree is an important exotic tree in Sudan which
contributes in forest sector as
shade, firewood, pesticide & medicinal
plant.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 22
NEEM & AGROFORESTRYNEEM & AGROFORESTRY
Neem has been selected to be among the best trees for Agro-forestry systems in dry & semi dry zones.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 23
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 24
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..
Jan. 05, 2007 Dr. Intisar E. Elteraifi 25
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 26
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 27
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 28
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 29
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 31
Neem Products
Jan. 05, 2007 Dr. Intisar E. Elteraifi 32
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 34
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 35
Pesticides hazards
Agricultural pests
Needs for More food
High cost
Sudan is an agricultural country
Jan. 05, 2007 Dr. Intisar E. Elteraifi 36
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 37
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 38
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 41
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 43
Climatological data & GPS
RF, Temp. , RH% and GPS data for the selected agro-ecological zones were obtained from the meteorological Department, Sudan.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 44
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 45
Annual rainfall and the selected zones in Sudan
6
12
8
43
9
75
10
Jan. 05, 2007 Dr. Intisar E. Elteraifi 46
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 47
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) .
Jan. 05, 2007 Dr. Intisar E. Elteraifi 48
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 49
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 50
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 51
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).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 52
Seedlings in block I were watered every 10 days, while the seedlings in block II were watered every day.
Block I Block II
Jan. 05, 2007 Dr. Intisar E. Elteraifi 53
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 54
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 55
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 56
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 57
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 58
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).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 60
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 62
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 63
Analytical High Performance Liquid Chromatography (Beckman HPLC, System Gold).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 64
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 65
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 66
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:
Jan. 05, 2007 Dr. Intisar E. Elteraifi 67
High performance liquid chromatography of the neem seed kernel methanol extract
Jan. 05, 2007 Dr. Intisar E. Elteraifi 68
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 69
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 70
KEY RESULTSKEY RESULTS
Jan. 05, 2007 Dr. Intisar E. Elteraifi 71
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 72
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 73
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).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 74
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 75
K.WT SH.WT
Contribution of the seed shell and seed kernel in the neem seed weight
Jan. 05, 2007 Dr. Intisar E. Elteraifi 76
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 77
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 78
Morphological variations in some neem provenances in Sudan. a) Seeds ; b) leaves ; c) roots.
a b
c
Jan. 05, 2007 Dr. Intisar E. Elteraifi 79
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 80
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 81
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 82
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 83
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 84
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 85
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 86
Effects of two irrigation regimes on neem seedlings from various ecozones (a) Irrigation I & (b) Irrigation II
a) b)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 87
Morphological variations between the two irrigation regimes in the root system of two seed sources: (a) S9 & (b) S3.
a) b)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 88
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 89
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).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 90
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).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 91
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 92
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.)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 93
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 94
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 96
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 97
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 99
Variations in nimbin level in different zones in Sudan over two seasons
Jan. 05, 2007 Dr. Intisar E. Elteraifi 100
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 101
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 102
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 104
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 106
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 108
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 112
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 113
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 114
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).
Jan. 05, 2007 Dr. Intisar E. Elteraifi 115
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 116
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 117
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 118
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
Jan. 05, 2007 Dr. Intisar E. Elteraifi 119
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 120
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 121
Jan. 05, 2007 Dr. Intisar E. Elteraifi 122
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 123
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.
Jan. 05, 2007 Dr. Intisar E. Elteraifi 124
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)
Jan. 05, 2007 Dr. Intisar E. Elteraifi 125
Future plansComing
back to India Karnataka
is interesting
for me
Jan. 05, 2007 Dr. Intisar E. Elteraifi 126