Taxonomy, Cultivation and Other Practices of Neem

1. Taxonomy Azadirachta indica A. Juss. Meliaceae

2. Distribution

The tree is found apparently wild on Siwalik hills. It grows wild in the dry forests of Andhra Pradesh, Tamil Nadu and Karnataka. Though not a forest-tree, it is generally found to grow wild. In Andhra Pradesh, most of the trees grow from self-sown seeds. It is cultivated all over India, especially in the drier parts of the country, and has evidently become wild in many localities as an escape from cultivation. The tree grows on almost all kinds of soils, including clayey, saline and alkaline soils, but does well on black cotton soils. It thrives better than most other trees on dry, stony, shallow soil with a waterless sub-soil, or in places where there is a hard calcareous or clay pan near the surface. It does not tolerate inundation. The tree increases the soil fertility and water-holding capacity, as it has a unique property of calcium-mining which changes the acidic soils into neutral. It is a very useful roadside and avenue-tree in dry and moderately dry climate and an excellent shade tree during the hot season when other trees are bare. It thrives, as a rule, where the maximum shade-temperature is as high as 49x, and the rainfall varies from 45 to 112 cm. The tree has been employed for afforestation of dry localities, for re-foresting bare ravines and for checking soil erosion, and also as windbreaks. It has also been used for mixed cropping [Troup, I, 179; Gamble, 143; Christopher, Indian Fmg, N S1, 20(1), 38; Katyal, ibid, 1956-57, 6(1), 36; Ketkar, Utilization of Neem (Azadirachta indica, A. Juss.) and its by-products, KVIC, Bomaby, 1976, 4; Lakshmikantan, First

Circular Symp Role Neem Indian Econ, New Delhi, 1972, No. NSI/RD/5(S)/7172, 1; Khan, 96; Mitra, C R, 3; Kaul, Indian For, 1957, 83, 375; Radwanski, World Crops, 1977, 29, 62; For Abstr, 1965, 26, 5116]. The tree is mostly evergreen, except in dry localities where it becomes almost leafless for a brief period during Feb-March and the new leaves appear during March-April. The flowering is spread over Jan- March in the Southern parts of the country and later towards the North. In Kerala, the flowering starts in the beginning of January; in Feb- March in Karnataka, Tamil Nadu and Andhra Pradesh; in the first week of April in central India getting progressively delayed in Northern India and Punjab, going up to first week of May in the sub-Himalayan areas. The tree often has a restricted bloom during Aug-Sept in the plains of Uttar Pradesh. The fruiting follows a similar pattern. The seed does not retain viability for long. Care is needed in collecting the seed: it should be collected from the tree when thoroughly ripe and sown as soon as possible after collection (Troup, I, 179; Mitra, C R, 3; Khan, 96).

The tree is a light-demander and in the young stages, pushes up vigorously through the scrubby jungle. It is hardy but frost-tender and does not withstand excessive cold, especially in the seedling and sapling stages. It coppices well and produces root suckers and withstands pollarding well. The young plants develop rapidly after the first season. The rate of growth is fairly good, showing five rings per 2.5 cm, giving a mean annual girth-increment of 3.2 cm (Troup, I, 180-83; Mitra, C R, 3; Gamble, 144).

3. Description

A large, evergreen tree, 12-18 m in height and 1.8-2.4 m in girth, with a straight bole and long, spreading branches forming a broad crown, commonly found throughout the greater part of India, and often cultivated. Bark gray or dark gray, rough, reddish brown inside with numerous oblique furrows and scattered tubercles; leaves imparipinnate, alternate, 20-38 cm long: leaflets 8-19, alternate or opposite, ovate- lanceolate, oblique or sub-falcate, falcate-lanceolate, glossy, bluntly serrate; flowers white or pale-yellow, small, scented, numerous, in long, slender, very lax, axillary panicles; drupes green, turning yellow on ripening, aromatic, oblong, or ovoid-oblong, smooth, 1.3-1.8 cm long, with a single exalbuminous seed.

The fruit is an ovoid, bluntly pointed, smooth drupe, green when young and unripe, yellow to brown when ripe, with a very thin picarp, mesocarp with scanty pulp and a hard bony endocarp, enclosing one seed.

4. Cultivation

Natural regeneration: Under natural conditions, the seed ordinarily falls to the ground during the rainy season and germination occurs within a week or two. It reproduces naturally with tolerable freedom (Troup, I, 180).

Artificial regeneration: Although the tree can be raised easily in the nursery and transplanted for afforestation purposes, direct sowings are reported to be more

successful than transplanting. For raising transplants and to protect the radicle from insects the seed should be covered with a thin layer of soil, and sparingly watered. The seedlings are fit for transplanting during the first rains, when they are 7-10 cm in height and the taproot is 15 cm long, if larger seedlings are required they may be retained for another year in the nursery and transplanted early in the rains of the second season. The stem and roots may also be pruned. In frosty localities, they are protected by screens. Weeding without water also helps in promoting growth. Loosening the soil to prevent caking and to promote soil aeration is also beneficial. It can also be propagated from root- and shootcuttings; application of growth-regulators (IBA and NAA) induces root formation (Troup, I, 180-83; Krishnaswamy, 1956, 61; Shanmugavelu, S. Indian Hort, 1967, 15, 70).

Propagation: Transplantation of stumps is well known in India. These are prepared from 2 years-old seedlings and are subsequently planted in 30 cm3 pits. Root-ball transplantation is another good method where one year-old seedlings are carefully uprooted along with a ball of soil around the roots and transplanted as soon as possible.

Apart from other techniques, tissue culture techniques have also been tried to propagate the plant and showed some encouraging results such as differentiation of growth centres in callus, root formation in MS medium containing IAA, and callus formation in some African progeny which may provide a basic for future research(Koul et al, Canad J Bot, 1990,68,1).

 To learn the best time for harvesting Neem (Azadirachta indica) seeds, the amount of the 5 major triterpenoids, together with the oil content were determined throughout a fruiting season in 6 selected trees in Sri Lanka. The triterpenoid content and the relative proportions of the major compounds changed little from the hard green fruit stage to mature seeds, while the oil content increased markedly with time. The highest content of azadirachtin (10 mg/g seed kernels) was recorded in newly ripened seeds. There was some loss of salannin and azadirachtin in storage after harvesting for up to 6 months. (Ann.Bot. (London), 78(3), 383,1996. Chem Abstr 125:243272).

Neem produces seeds in rainy season. Phenotypes that produced seeds outof-season in Nov.-Dec. were identified. Seeds from the normal and out-of-season (winter) were screened for quality and quantity variations in azadirachtin content. Monsoon seeds yielded higher (1.53%) azadirachtin-rich fraction as compared to the winter season seeds (1.26%). Azadirachtin A, B and F were analyzed by HPLC using an improved solvent-eluting system for better resolution. Azadirachtin A was the major metabolite in the rainy season seeds, as compared to azadirachtin B and F, however, azadirachtin A and B were in nearly equal proportions in the winter seeds. Concentration of azadirachtin F increased more than two-fold in winter seeds as compared to that of rainy ones. The sum of the peak area of azadirachtins A, B and F was higher in the seeds produced in winter. Winter stress appears to favor synthesis of azadirachtin B and F. Such phenotypes could be a material of choice for clonal multiplication of neem. ( Curr. Sci., 70(12), 1084,1996. Chem Abstr 125,163284)

The hairy root cultures of Azadirachta indica were initiated by co-cultivation with Agrobacterium rhizogenes MTCC 532 using modified MS medium

supplemented with 500 mg/l of cefatoxime without plant hormones. The hairy roots were extracted for azadirachtin which was found to be 1.910% on dry wt. basis by HPLC analysis. (Indian Drugs, 35(8), 485, 1998; Chem Abstr 129, 289208)

Somatic embryos were initiated with mature seeds of neem ( Azadirachta indica) when cultured on Murashige and Skoog's medium supplemented with thidiazuron (TDZ). Regeneration occurred via somatic embryogenesis: direct embryo formation and through an intermediary callus phase. TDZ was very effective and induced somatic embryogenesis across a wide range of concentrations. (1-50 mM). However, somatic embryogenesis was accompanied by callus formation at concentrations of 20 mM and above. Cell suspension cultures were established with the TDZ-induced callus and groups of large cell clumps were formed within 2-3 wk. Plants were regenerated from both directly formed somatic embryos and somatic embryos derived from cell suspensions plated on semisolid medium devoid of growth regulators. Regenerated plantlets continued to grow after transfer to a greenhouse environment and were similar phenotypically to zygotic seedlings. This simple regeneration system may be beneficial for mass propagation of selected elite clones of neem. ( Plant Cell Rep., 17(6-7), 469,1998 Chem Abstr 128,268336)

Micropropagated shoots were initiated from leaf explants of the neem tree, Azadirachta indica. Regardless of their origin, shoots were successfully produced by culturing leaf explants on Murashige and Skoog medium containing benzylaminopurine (1 mg/L), kinetin (0.8 mg/L) and adenine sulfate (6 mg/L) in complete darkness. These shoots were further multiplied on Murashige and Skoog medium containing benzylaminopurine (0.1 mg/L), kinetin (0.08 g/L) and

adenine sulfate (0.6 mg/L). Within 32 wk, 80 shoots could be produced from a single leaf explant (10 mm 10 mm). 55% Of these shoots rooted on Murashige and Skoog medium containing indolebutyric acid (1 mg/L) and all of these grew on transfer to soil. (Plant Cell Rep., 17(3), 215, 1998; Chem Abstr 128, 86548)

Role of permeabilizing agents on secretion of azadirachtin from margosa callus cultures was investigated under laboratory conditions. The results indicated that most of the synthesized azadirachtin remains inside the cells. Different permeabilizers showed variation in secretion pattern. The most effective was found to be Triton X-100, inducing azadirachtin secretion of 10 mg/L at 150 ppm, followed by DMSO and chitosan. This is the first report of its nature on permeabilizing agent induced enhanced secretion of azadirachtin from neem callus cultures. (Indian J. Exp. Biol.,37(1), 89, 1999 Chem Abstr 130:264992)

Callus cultures of A. indica leaves and flowers were established for the production of azadirachtin. The 12-wk old flower callus produced 2.46% of azadirachtin. (Fitoterapia, 69(5), 423, 1998 Chem Abstr 130:167210)

From immature embryos of Azadirachta indica (A. Juss), callus was developed with 100% response on MS medium supplemented with (a) 2.0 mg L-1 NAA + 0.5 mg L-IBA, (b) 2 mg L-I BA and (c) 1.0 mg L-IAA + 0.5 mg l-IBA. For optimum multiple shoot formation, medium containing (a) 2.0 mg L-IBA and (b) 0.5 mg LIBA + 0.1 kgl-1 NAA was used which rooted in 1/2 strength MS medium supplemented with 0.5 mgl-1 IBA. Azadirachtin and nimbin content were analyzed in callus and in cultured roots and shoots. Roots had 0.004 mg/g

azadirachtin, and shoots had 0.008 mg/g azadirachtin and 0.003 mg/g nimbin. Callus had negligible amt. of azadirachtin. (Indian J. Plant Physiol., 3(2), 129, 1998 Chem Abstr 130,136561)

The influences of different explants from organs of Azadirachta indica, culture media and subculture times on callus growth and azadirachtin content in cultures were studied. The results showed that all of the call induced from root, leaf, stem and bark had the capability of synthesizing azadirachtin, among which callus growth rate and azadirachtin from leaf explants were the highest. B5 medium with low ammonium salt had good result for callus growth, while MS medium with high ammonium salt was beneficial for azadirachtin accumulation, but White medium without ammonium salt was satisfactory for neither of them. The best callus growth and azadirachtin formation occurred at the second or third subculture. 130:280876) (RedaiYaredai Zhiwu Xuebao, 6(3), 267,1998 Chem Abstr

Azadirachta indica branches air layered in July to September showed 90-100 percent rooting, while in those of October and November the percentage of rooting was 30-40 percent. Airlayering has been reported as the best method for multiplication of neem tree in monsoon season. (Med. Arom. Plant. Abst., 21(4), 9904-2155,1999; Palanisamy, K.., Indian Forester, 125(3), 331, 1999)

From immature embryos of Azadirachta indica callus was developed with 100 percent response on MS medium supplemented with 2.0 mg/1 NAA + 0.5 mg/1 B A, 2 mg/lBA and 1.0 mg/lAA + 0.5 mg/lBA. For optimum multiple shoot formation, medium

containing 2.0 mg/lBA and 0.5 mg/lBA+0.1 mg/1 NAA was used which rooted in 1/2 strength MS medium supplemented with 0.5 mg/lBA. Azadirachtin and nimbin content were analysed in callus, in vitro cultured roots and shoots. Roots had 0.004 mg/g azadirachtin and shoot had 0.008 mg/g azadirachtin and 0.003 mg/g nimbin. Callus had negligible amount of azadirachtin.. (Srividya, N. et al; Indian Journal of Plant Physiology, 3(2): 129, 1998 )

The best callus growth was achieved with leaf explants cultured on modified MS medium supplemented with 2,4-D-kinetin and glycine. Modified MS medium with NAA and kinetin, was found to be suitable for flower callus. The 12 weeks old flower callus and 20 weeks of leaf callus 2.46 and 2.68 percent of azadirachtin respectively. (Veeresham, C et.al.; Fitoterapia, 69(5), 423,1998 )

The effect of auxins (IAA, IBA), B vitamin (thiamine) and boric acid on rooting response of stem cuttings of Azadirachta indica in relation to different seasons was investigated. The cuttings collected from a tree showing complete leaf fall or bud breaking stage in the month of February developed significant adventitious-roots and in remaining seasons the rooting percentage was very poor, 1000 ppm IBA was found to be the best treatment which induced 80 percent rooting in February. The effect of auxins or vitamin on root stimulation was found to be dependent upon the physiological and cambial activity of the cuttings and was seasonal. (Palanisamy, K. and Pramod Kumar ; Indian Journal of Forestry, 19(2), 183, 1996)

 Forty-two neem ecotypes of India have been found to show a wide variation in the content of oil, and their physicochemical characteristics, total fatty acid, fatty acid composition, and the key meliacins (azadirachtin, nimbin, and salannin). The azadirachtin content did not correlate with any of the physicochemical and chemical parameters, but the nimbin and salannin contents correlated significantly with each other. The refractive index and iodine value showed weak but significant correlation with the contents of nimbin and salannin. Insect growth inhibition of Spodoptera litura revealed a wide variation in the EC50 of the oils. The salannin and azadirachtin contents of the oils correlated the most with bioactivity. (Kumar, J. and Parmar, B.S. Journal of Agricultural and Food Chemistry, . 44(8): p. 2137-2143, 1996).

Wood anatomy, macro and microscopic study and chemical tests of powdered drug of Zanthoxylum armatum and Azadirachta indica used as miswaks were done. Z.armatum wood shows vessels, rare parenchyma, and abundant fibres while A.indica wood showed vessels, parenchyma, ray cells, growth ring and pith surrounded by larger vessels. Powdered miswak of both showed fibres, parenchyma, and fragments of vessels. Powder of Z.armatum was completely insoluble in sulphuric acid, hydrochloric acid, acetic acid, benzene and water while A.indica powder was insoluble in sulphuric acid and hydrochloric acid. Powdered miswaks of both species lost their original colours in various solvents. Both appeared to be safe for human teeth. (Dastagir, G. and Haq, I.U; Journal of Science and Technology, University Peshawar, 21(1&2), 57, 1997).

5. Diseases & Pests

The tree is not subject to serious fungal diseases. Fungi recorded on the tree include Cercospora leucosticta Ellis & Everh., C. subsessilis Syd., Fomes senex Nees & Mont., Polyporus gilvus Schw. and Xylaria azadirachta Anahosur. Although many pests have been recorded, the tree is not attacked by any serious pest; in some localities porcupines damage the trees [ Butler et al, 182, 258, 260; Sarbhoy et al, 69; Indian J Agric Sci, 1950, 20, 126; Joshi, Agra Univ J Res (Sci), 1957, 6(2), 15].

Six fungal genera representing Fusarium, Aspergillus, Penicillium, Alternaria, Cephalosporium and Pythium, apart from Mycelia sterelia and bacteria were isolated from seeds of Azadirachta indica. Among these the dominant species were Aspergillus niger, A.fumigatus and Fusarium solani. (Uniyal, K. and Uniyal, D.P.; Indian Forester, 122(5), 423, 1996).

Neem cake, neem leaves (powder or aqueous extract) neem oil and extracts of neem seed or kernels, have been found to control phytopathogenic fungi on many crops such as tomatoes, egg plant, rice, cotton, soya, grapes, wheat, beans, roses and cucumbers. Neem (Azadirachta indica) seed extract is very effective in controlling Plasmopara viticola, an important pathogen of grapes and Sphaerotheca fuliginea, a pathogen of cucumbers. (Steinhauer, B.; Plant Research and Development, 50, 83, 1999).

 The effect of host plants like castor, cabbage, cauliflower, tomato and wild cabbage and also of neem oil on food utilization indices of Spodoptera was observed. Cauliflower was found to be the most preferred host as all three food utilization indices like ECI (24.13 percent), AD (50.54 percent) and BCD (34.86 percent) were significantly higher as compared to the larvae fed on other host plants. The neem (Azadirachta indica) oil treated leaves of cauliflower when fed to larvae recorded significant lower food utilization indices as compared to untreated plants, (Sharma, B.L. et.al.; Annals of Agri-Bio Research, 4(1), 37, 1999).

Azadirachta indica leaf leachate was prepared by immersing 20 gm seeds in one litre of water for 48 hours. The solution was filtered and used as leachate. 50 seeds of each crop were grown in plastic boxes and irrigated with water (as control) and leachate. After 10 days observations were made and revealed that leaf extract had no effect on germination and root growth in crops except mustard where rate of germination decreased. Shoot length of all crops except pearlmillet showed marked decrease when irrigated with leachate. This indicates that allelochemicals of leachate may be stimulatory to some crops and inhibitory to some. In this case pearlmillet may prove most compatible crop with Azadirachta. ( Haryana Journal of 'Agronomy, 12(1), 75, 1996).

Other Practices

 The sapwood is grayish white; heartwood is red when first exposed, fading to reddish brown. The wood is dull to somewhat lustrous, especially on the radial surface, aromatic but without any characteristic taste, moderately heavy (wt, 705945 kg/cu m), rather uneven and narrowly interlocked-grained and medium- to somewhat coarse-textured. The wood is durable even in open situations. It is not usually attacked by insects. The timber is medium refractory and seasons well, even when converted from green logs. Boards, cut from green logs, should be seasoned in open stacks placed under cover. It is not difficult to saw and in this respect is about equal to teak. It is very easy to work by hand and on machines and turns on a lathe to a fair finish and lends itself for broad carving, but does not take polish well. The data for the comparative suitability of the wood, expressed as the percentages of the same properties of teak, are: wt. 124; strength as a beam, 87; stiffness as a beam, 81; suitability as a post or strut, 82; shock-resisting ability, 105; retention of shape, 77; shear, 129; and surface-hardness, 131; nail- or screw-holding property, 117 [Pearson & Brown, I, 235-37; Gamble, 144; Sekhar & Gulati, Indian For Rec, N S, Timb Mech, 1969, 1(1), 25].

The wood is used for furniture, carts, axles, yokes, naves and felloes, boards and panels, cabinets, bottoms of drawers, packing- cases, ornamental ceilings, ship- and boat-building, helms, oars, oil- mills, cigar boxes, carved images, toys, drums and agricultural implements. It is suitable for timber-bridges up to 5 m span. Trunks and chests are made out of this wood and are pest-proof (Pearson & Brown, I, 237; Mitra, C R, 5; Dastur, Useful Plants, 39; Lewis, 92; Masani & Bajaj, Indian For, 1962, 88, 750).

 The pulp of the fruit is eaten by humans, birds and animals. The pulp water when sprayed, protects crops from locusts (Mitra, C R , 9; Radwanski, World Crops, 1977, 29, 62; Information from the All-India Non-Edible Oil Industry Association, Pune).

It is estimated that India has about 13800000 neem trees with the potential to produce over 83000 tonnes of neem oil and 330000 tonnes of neem cake from 413000 tonnes neem seeds. Single neem tree yields 37-50 kg fruits per year. Forty kilograms of fresh fruits yield nearly 24 kg of dry fruits (60%), which in turn give 11.52 kg of pulp (48%), 101 kg of seed coat (45%), 6.0 kg of husk (25%) and 5.5 kg of kernel (23%). The kernel gives about 2.5 kg of neem oil (45%) and 3.0 kg of neem cake (55%). It has been reported that a 50 years old tree yield @ 51 kg fuel wood in arid areas of Rajasthan. The neem wood is generally considered to be highly resistant to fungi and insect attack and is durable even when used outdoors (Koul et al, Canad J Bot, 1990, 68, 1).

Margosa tree (Azadirachta indica) has been universally accepted as a wonder tree because of its multiple uses. The tree has been considered so valuable and miraculous that it became a major component of the Indian ecosystem. Medicinal uses of neem are known since Vedic times and all parts of tree have been in use especially in Ayurvedic and Unani system of medicine. Besides, neem also holds unlimited potential as a pesticide and agrochemical. The botany of neem and its potential for use all over the globe have been summarised., Botanica, 47, 58, 1997)

Seed morphology (seed length and 20 seed weight) and oil content was studied in Azadirachta indica of five provenances of Northern and Western India. Trees with wide

ranges of girths were considered for study. Maximum average oil content was observed in trees from Hisar provenance. Seed oil content in most of the provenances was not consistently and significantly correlated with morphological parameters of seeds. Age of the tree had no significant effect on the oil yield. (Kaura, S.K. et.al., Plant Food for Human Nutrition, 52(4), 293, 1998)