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| Biomass Research Station |
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| Group Leader |
Dr. H. M. Behl, Sci 'G' |
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Contact |
Work
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91-522-2205842 Ext.
303
91-522-2205847 |
| Home |
91-522-2763108 |
| Fax |
91-522-2205847 |
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Other Scientists of the Group
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Dr. V. L. Goel, Scientist EI
Dr. Nandita Singh,Scientist C
Dr. O. P. Sidhu, Scientist B
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Introduction
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| Azadirachta
indica A. Juss. (Neem) of family Meliaceae is native
to the Indian subcontinent. Neem, today, is grown in many
Asian countries and in tropical regions of the western hemisphere.
Leaves and seeds of this tree have traditionally been used
for centuries for treatment of human ailments and control
of pests. Azadirachtin (C35H44O16),
a tetranortriterpenoid from neem kernel has been rated as
the most potent naturally occurring insect feeding deterrent
and has generated wide academic and industrial interests.
Azadirachtins function as natural insect control agents
because of their antifeedant as well as insect growth regulatory
properties. Several active metabolites in various plant
parts of neem bestow it unique properties as health protective,
promotive, curative properties apart from it being a pesticide
par excellence, universally accepted biofertilizers and
a major constituent of village and household dispensaries3. |
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Figure
1: Neem tree with high seed output |
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Background
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NBRI was involved in neem research since early 1970s
when Dr. Mitra wrote perhaps the first book on Neem detailing various
chemical constituents, several of which were unknown at that time.
However, neem was not further explored in the following years. Neem
plantation trials were laid at Biomass Research Center of the institute
in 1988 but the investigations were not intensive. Dr. H. M. Behl
proposed a national program on neem in mid 1990s with the encouragement
of (late) Professor S. K. Sinha the then Chairman of the Research
Council of Institute could not proceed due to lack of sufficient
funding. In 1999 National network programme involving 11 institutions
in country was launched in India with NBRI as the nodal Institution
and Dr. H. M. Behl as the coordinator of the same. Further development
of technologies and products was initiated in 2000, based on research
findings under this programme. Currently, NBRI maintains a large
number of accessioned germplasm with reference material. Also products
and technologies have been developed for commercial exploitation.
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R&D Program and major achievements
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Neem exhibits a large variability as s it grows in
a variety of habitats and in almost all states of the country. There
is often a mention in literature about variations in its major metabolites
such as azadirachtin content in neem from different countries or
different regions within a country. Most of the studies using chemical
markers and molecular biology tools had been contradictory.
The Biomass Biology group undertook extensive surveys from throughout
the country, more extensively from Bihar and Uttar Pradesh and screened
the germplasm for phenological and phytochemical traits. Other colleagues
(Dr. Vijay Laxmi Goel and Dr. Nandita Singh) provided valuable inputs
in collection, accessioning and evaluation of the germplasm)1.
Various provenances of neem were screened for oil percentage, azadirachtin
A and B, salanin and nimbin contents. The metabolites were extracted
using conventional methods and analyzed by High Pressure Liquid
Chromatography (HPLC). Earlier Sidhu and Behl (1996) have developed
an effective HPLC protocol for detection of azadirachtin A, B and
F5. Nimbin and salanin were quantified using RP-analytical
HPLC. A large variability was observed in azadirachtin content (Figure
2) Azadirachtin A varied from 556.9 to 3030.8 mg kg-1
with an average of 1327.4 mg kg-1 amongst various provenances.
Certain individual trees (IC268646) within a provenance had only
negligible amount of (68.5 mg kg-1) azadirachtin A. Even
within a provenance there was a great variability in azadirachtin
A content. No correlation between azadirachtin A and B could be
established6.
It was observed that ratio of A to B may be from as low as 1.12
to a high of 98.1. The studies revealed that azadirachtin A is usually
13 to 16 times higher than azadirachtin B but wide individual differences
were observed among natural population.
Average of maximum and minimum temperatures, relative humidity and
rainfall at different provenances were monitored throughout the
year during the study.
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Figure 2: Variability in
azadirachtin A and B
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Low oil percentages were observed from regions under
stress of aridity, salinity or alkalinity. Jodhpur, Jaisalmer, Ajmer
in the dry zone with low humidity and high summer temperature, and
Kanyakumari with a stress of salinity had less than 35 % oil. These
areas also had a low ratio of azadirachtin A to B. Jaisalmer, for
example is an extreme arid site. It had 34.5 % oil and A:B ratio
was 2.56. Kanyakumari, located at the southern tip of landmass,
was in the similar range. These are only trends since there were
exceptions and no significant correlation between oil and azadirachtin
content was observed. Kanyakumari that had least variability in
temperature and humidity throughout the year also had least variability
in azadirachtin A, azadirachtin B content and their ratios. No such
conclusion could be drawn at other areas where there is a large
variability in daily and seasonal climatic conditions.
Elite individuals with exceptionally higher salanin and nimbin content
were identified (Figure 3). There were wide variations in seed area,
oil content, nimbin and salanin concentrations among different provenances
and also between individual trees of a particular provenance7.
Nimbin concentration ranged from 18.2 to 636.8 mg kg-1
whereas salanin was in the range of 45.4 to 1830.3 mg kg-1
kernel weight. Analysis of variance for seed area, oil content,
nimbin and salanin among various neem provenances showed significant
differences in oil content, nimbin and salanin. Correlation coefficients
among the parameters investigated showed nimbin and salanin as significantly
correlated. Nimbin and salanin content also varied between individual
trees of a particular provenance.
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| Figure 3: Variability
in salanin and nimbin content
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The study shows that nimbin or salanin synthesis
did not appear to be influenced by environmental conditions as individual
trees from the same agro climatic zone showed different trends.
It can be concluded that there are individual genetic differences
among neem trees.
Hasty conclusions of proposing a particular country or provenance
yielding high azadirachtin may be risky. Findings based on small
sample size or conclusions based on a multitude of factors can be
misleading. With nearly 70 countries in the world interested to
raise neem plantations, it is crucial that issues regarding variability
in neem are understood. It can be concluded from present study that
there are individual genetic differences among neem trees and synthesis
of azadirachtin is not dependent upon temperature, humidity or rainfall.
Genetic variability for azadirachtin (Tables 1, 2) may affect the
final products. Large variability provides ample opportunity for
selection and further improvement. It may not be feasible at this
stage to select a particular individual with desired traits as no
selections have been made in neem so far. A systematic study for
tree improvement with a population of mother trees with desired
traits should be undertaken by laying half-sib progeny trials and
further selections by clonal propagations2. The role
of genetic make up needs further research. |
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Table 1: Seed area, oil content and azadirachtins
(A & B) among neem accessions from different agro climatic zones
of India
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Provenance |
Seed Area (mm2) |
Aza (A) mg/kg Kernel wt. |
Aza (B) mg/kg Kernel wt. |
Total A+B |
A:B ratio |
Oil (%) |
| 1 |
Orai1 |
83.5 |
556.9 |
225.7 |
782.6 |
2.47 |
44.7 |
| 2 |
Chapra2
|
98.1 |
678.6 |
111.5 |
790.2 |
6.13 |
39.7 |
| 3 |
Auraiya1
|
83.7 |
804.2 |
68.1 |
872.3 |
33.07 |
37.1 |
| 4 |
Kanyakumari3
|
88.9 |
829.9 |
379.4 |
1209.3 |
2.95 |
35.0 |
| 5 |
Bijnor1
|
73.4 |
834.8 |
177.1 |
1011.9 |
4.71 |
46.2 |
| 6 |
Kannauj1
|
102.1 |
880.2 |
312.3 |
1192.5 |
3.32 |
36.4 |
| 7 |
Rampur 1
|
72.3 |
929.7 |
88.9 |
1018.7 |
10.48 |
37.3 |
| 8 |
Gazipur1
|
98.8 |
942.6 |
66.2 |
1008.9 |
14.31 |
42.7 |
| 9 |
Ajmer4
|
86.3 |
955.5 |
316.2 |
1271.7 |
3.02 |
33.2 |
| 10 |
Bhagalpur2
|
100.3 |
980.6 |
101.9 |
1082.5 |
9.62 |
43.6 |
| 11 |
Barauni2
|
90.9 |
982.7 |
147.4 |
1130.1 |
7.25 |
41.8 |
| 12 |
Darbhanga2
|
89.9 |
1002.5 |
119.7 |
1122.1 |
8.42 |
45.5 |
| 13 |
Jodhpur4
|
68.8 |
1003.1 |
103.2 |
1106.3 |
9.71 |
32.9 |
| 14 |
Moradabad1
|
101.3 |
1013.3 |
214.7 |
1228.1 |
6.41 |
50.1 |
| 15 |
Farrukhabad1
|
112.3 |
1022.5 |
94.0 |
1116.5 |
10.87 |
42.5 |
| 16 |
Mathura1
|
90.5 |
1071.8 |
58.7 |
1130.5 |
31.41 |
45.3 |
| 17 |
Kheri1
|
91.6 |
1082.6 |
173.2 |
1255.7 |
5.88 |
43.1 |
| 18 |
Jaisalmer4
|
75.2 |
1092.2 |
426.2 |
1518.4 |
2.56 |
34.5 |
| 19 |
Hathras1
|
82.4 |
1126.9 |
134.4 |
1261.2 |
8.40 |
42.4 |
| 20 |
Ranchi7
|
92.5 |
1159.8 |
190.1 |
1349.9 |
6.09 |
41.2 |
| 21 |
Muzzafarpur2
|
90.1 |
1169.4 |
141.5 |
1310.9 |
8.26 |
41.6 |
| 22 |
Bulandshahar1
|
97.0 |
1205.1 |
139.4 |
1344.5 |
9.79 |
49.7 |
| 23 |
Haridwar5
|
83.3 |
1231.0 |
182.8 |
1413.8 |
7.17 |
46.1 |
| 24 |
Hazipur2
|
101.1 |
1254.4 |
96.6 |
1351.0 |
13.04 |
42.9 |
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| 25 |
Patna2
|
86.2 |
1272.6 |
228.3 |
1500.9 |
5.58 |
41.1 |
| 26 |
Lucknow1
|
105.2 |
1334.2 |
139.4 |
1473.6 |
9.66 |
45.2 |
| 27 |
Badaun1
|
97.8 |
1388.1 |
143.9 |
1532.0 |
9.64 |
35.2 |
| 28 |
Dehradun5
|
89.3 |
1459.7 |
326.6 |
1786.3 |
4.72 |
45.3 |
| 29 |
JP Nagar1 |
74.9 |
1505.2 |
590.6 |
2095.9 |
6.03 |
37.1 |
| 30 |
Varanasi1
|
104.8 |
1531.7 |
69.7 |
1601.5 |
24.54 |
48.3 |
| 31 |
Muzzafarnagar1
|
88.8 |
1534.5 |
149.7 |
1684.2 |
10.27 |
40.1 |
| 32 |
Meerut1
|
101.2 |
1573.8 |
105.2 |
1679.0 |
22.63 |
40.8 |
| 33 |
Agra1
|
76.2 |
1576.2 |
261.9 |
1838.0 |
8.16 |
43.9 |
| 34 |
Siwan2
|
81.1 |
1629.0 |
96.0 |
1725.0 |
17.0 |
42.8 |
| 35 |
Kanpur1
|
80.5 |
1671.4 |
68.9 |
1740.4 |
23.14 |
40.8 |
| 36 |
Pratapgarh1
|
78.9 |
1705.9 |
100.8 |
1806.7 |
17.12 |
38.6 |
| 37 |
Jaipur4
|
92.6 |
1706.4 |
251.5 |
1957.9 |
6.78 |
33.5 |
| 38 |
Samastipur2
|
84.5 |
1916.5 |
95.0 |
2011.5 |
20.19 |
45.0 |
| 39 |
Fatehpur1
|
97.0 |
1941.2 |
43.1 |
1984.3 |
98.15 |
51.6 |
| 40 |
Rohtak6 |
86.6 |
2032.9 |
243.9 |
2276.8 |
8.33 |
37.9 |
| 41 |
Mau1 |
92.6 |
2058.2 |
100.3 |
2158.5 |
20.53 |
43.2 |
| 42 |
Sri Ganganagar4
|
89.0 |
2400.0 |
172.3 |
2572.3 |
13.92 |
37.3 |
| 43 |
Shahjahanpur1
|
110.9 |
3030.8 |
134.1 |
3165.0 |
24.32 |
40.6 |
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CD at 1% |
61.10 |
1023.04 |
326.10 |
1126.3 |
33.87 |
4.45 |
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CD at 5% |
45.94 |
769.20 |
245.19 |
846.8 |
25.47 |
3.34 |
|
| 1 = Uttar Pradesh; 2 = Bihar; 3 = Tamil Nadu; 4=
Rajasthan; 5 = Uttranchal; 6 = Haryana; 7 = Jharkhand. |
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| Table 2: Analysis of variance between seed
area, oil content, azadirachtin A, B; A+B and A:B ratio among different
provenances |
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| Source |
Degree of Freedom |
SS |
MS |
F value |
| Seed area (mm2) |
42 |
11446.1 |
317.9 |
0.4 NS |
| Oil (%) |
42 |
1393.97 |
38.7 |
9.2** |
| Azadirachtin A |
42 |
2.31 x 107 |
642225.1 |
2.9* |
| Azadirachtin B |
42 |
1168336 |
32453.7 |
1.4 NS |
| Total Azadirachtin
(A+B) |
42 |
2.32 x 107 |
647116.4 |
2.4* |
| A:B ratio |
42 |
28426.2 |
789.6 |
3.2* |
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| Results are of two-way ANOVA (seed parameters, oil
content, azadirachtin A, azadirachtin B, total azadirachtin (A+B),
A/B ratio, and various provenances); total=36; * =P<0.01; **
=P<0.001; NS =not significant. |
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| Dental Cure Recipe from Neem |
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The authors developed a unique recipe for dental
cure. It filed a patent (Non toxic Dental care herbal formulation
for preventing dental plaque and gingivitis Patent Application No.
569 NF 2002)4.
Neem extracts reduce gingival bleeding and inflammation. These are
also useful against plaque formation. Neem extracts and the prouct
developed with synergistic effect from the herbs eliminated anaerobic
micro-organisms (Peptococcus, Fusobacterium and
Bacteroides) and aerobic species (Streptococcus viridans,
Bacilli, E. coli and Streptococcus epidermidis) within
a few days after application. |
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| Skin care, pet care and other products |
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The group has developed several products like skin
care, pet care, pesticidal formulations, fertilizers etc. from various
parts of neem involving state of art technologies.
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International Conferences
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Dr. H M Behl was the Chief Editor of Proceedings
of the World Neem Conference (Neem 99) held at Vancouver, British
Columbia, Canada. The proceedings were published by the Neem Foundation,
India. He was also the co-editor of proceedings entitled “Collection,
Processing and Commercial Utilization of Neem”, published by the
the Ministry of Industries, Govt. of India in 2000.
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Acknowledgments
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The study was a part of National Neem Network activities.
The network was sponsored by the NOVOD Board, Ministry of Agriculture,
Govt. of India.
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Literature cited
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- Behl, H.M. (2000). Collection, Processing and Commercial Utilization
of Neem. Published by the Fragrance & Flavour Development
Centre, Ministry of Industries, Govt. of India, 2000; Co-editor:
A. Lahri.
- Behl, H.M. (2000). Technical manual for High Tech Protected
Environment Nursery Cultivation, National Botanical Research
Institute publication, Lucknow, India.
- Behl, H.M. (2002). Proc. World Neem Conference (Neem 99),
Vancouver, British Columbia, Canada, Published by Neem Foundation,
India.
- Behl, H.M., Saimbi, C.S., Srivastava, N. and Kant, S. (2000).
In: Collection, Processing and Commercial Utilization of Neem
(Eds. A. Lahri, H. M. Behl), Fragrance & Flavour Development
Centre, Govt. of India, pp. 239-242.
- Sidhu, O.P. and Behl, H.M. (1996). Current Sci. 70(12):
1084-1086.
- Sidhu O.P., Kumar, V. and Behl, H.M. (2003). Industrial Crops
& Products, 2003.
- Sidhu O.P., Kumar, V. and Behl, H.M. (2003). J. Agr. Food
Chemistry, 51(4):910-915.
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