• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• The Korean Journal of Zoology
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• v.32 no.4
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• pp.441-449
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• 1989

Azadirachtin은 Leucophaeq muderae (Fabricius연 하루 활동주기길이를 0.16 $\pm$ 0.03 h 단축시켰으며, 실험동물의 40 %에서 리듬의 Splitting현상이 관찰되었다. 하루에 두개의 활동주기를 보이는 Splitting현상은 서로 180$^{\circ}$를 유지하여 나타났고, 이는 동물의 실험 전의 본래 주기길이가 평균보다 더 긴 주기를 지닌 동물에게 주로 나타났다. 또한 활동주기의 Splitting현상을 보이는 동물의 안구와 뇌신경(optic lobe) 절단수술 후 Splitting이 사라짐으로써 Azadirachtin이 뇌신경에 자극을 주어 Splitting을 야기시켰으며, 한쌍의 optic lobe 중 어느 한쪽만으로도 동물의 활동주기 조절가능성이 있다는 것이 본 실험을 통하여 밝혀졌다. Azadirachtin shortens the period length of the locomotor activity rhythm in the circadian rhythm of Leucophoeo maderae and induces splitting of this rhythm in ho components in about 40% of the animals. The phase relationship between the ho components is 180$^{\circ}$Both shortening of period and splitting are more pronounced in animals processing longer periods before the injection of azadirachtin. Splitting disappears after the surgical transection of optic lobe.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.4
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• pp.185-193
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• 2002

With Increasing awareness towards environment-friendly and non-toxic pesticide azadirachtin obtained from neon tree (Azadirachta indica) is gaining more and more importance. Its broad-spectrum activity, Peculiar mode of action. eco-friendly and non-toxic action towards beneficial organisms has offered many advantages over chemical pesticides. All currently use commercial formulations based on azadirachtin contains azadirachtin extracted from seeds of naturally grown whole plants which is labour intensive process depending upon many uncontrollable geographical and climatic factors. Plant tissue culture can be a potential process for the pro-duction, offering consistent, stable and controlled supply of this bioactive compound, However the research on tissue culture aspects of production are in preliminary stage and requires culture and process optimization for the development of a commercially viable process. This review states the present status and future challenges of plant tissue culture for azadirachtin production.

• The Korean Journal of Pesticide Science
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• v.13 no.4
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• pp.315-324
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• 2009

For the environmental friendly management of diamondback moth, Plutella xylostella (L.), Bacillus thuringiensis subsp. kurstaki ($30{\times}10^8\;cfu/mg$) and neem oil (0.5% azadirachtin) were used as green control agencies with mixed and alternative treatments on the chinese cabbage. When Bacillus thuringiensis subsp. kurstaki was applied to 1st and 2nd larva of P. xylostella with recommended concentration, their mortalities were reached to 100% by 2 days after treatment. In case of azadirachtin, its effect of mortality was continued for 7 days, and reached to 100% mortality. $LC_{50}$ values of Bacillus thuringiensis subsp. kurstaki and azadirachtin against 1st, 2nd, 3rd and 4th larva of P. xylostella were $2.8{\times}10^4$, $3.1{\times}10^4$, $3.4{\times}10^4$ and $1.5{\times}10^5\;cfu/ml$, and 2.7, 3.9, 4.7 and 7.1 ppm, respectively. The number of laid eggs of P. xylostella was reduced to 57.5 at 25 ppm of azadirachtin compared with control treatment. The hatch ratio was not significantly different with Bacillus thuringiensis subsp. kurstaki treatment in comparison with control treatment. However, when azadirachtin was applied, their hatch ratio were reduced to 25.8 and 45.4% at 25 and 50 ppm, respectively. On the other hand, emergence rate of eggs was not different with Bacillus thuringiensis subsp. kurstaki treatment, but 45.4% was shown in azadirachtin treatment with 50 ppm in comparison with control. When the mixture with Bacillus thuringiensis subsp. kurstaki and azadirachtin was applied to adults of P. xylostella, their mortality was higher than Bacillus thuringiensis subsp. kurstaki treatment only. These results are supposed that the mixture of Bacillus thuringiensis subsp. kurstaki and azadirachtin might be used as green control agents for reducing the demage of diamondback moth in the Chinse cabbage.

• Natural Product Sciences
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• v.25 no.2
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• pp.150-156
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• 2019

Conventional extraction of oil and azadirachtin, a botanical insecticide, from Azadirachta indica involves defatting the seeds and leaves using hexane followed by azadirachtin extraction with a polar solvent. In order to simplify the process while maintaining the yield we explored a binary extraction approach using Soxhlet extraction device and hexane and ethanol as non-polar and polar solvents at various ratios and extraction times. The highest oil and azadirachtin yields were obtained at 6 h extraction time using a 50:50 solvent mixture for both neem leaves (44.7 wt%, $720mg_{Aza}/kg_{leaves}$) and seeds (53.5 wt%, $1045mg_{Aza}/kg_{leaves}$), respectively.

• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.121-129
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• 2003

Callus cultures of Azadirachta indica flower petals were established on MS medium supplemented with naphthalene acetic acid (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$. Cell cultures of Azadirachta indica were established and studied the growth and production kinetics. Half 85 medium supplemented with dicamba (2 mg/L), kinetin (1 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable for initiation and maintenance of cell cultures from the calli. MS medium supplemented with naphthalene acetic acid (NAA) (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable as production medium. Around $80\%\;(0.05\%\;w/v)$ of azadirachtin was found to be intracellular. The effect of various precursors, elicitors, permeabilizing agents and growth retardants in cell cultures was studied. The addition of precursors sodium acetate (10 mg/L), squalene (10 mg/L), isopentenyl pyrophosphate (1 mg/L) and geranyl pyrophosphate (1 mg/L) to the cell cultures on day 3 has shown significant increase in bioproduction of azadirachtin $(64.94{\pm}4.40\;mg/L,\;72.81{\pm}0.04\;mg/L,\;51.63{\pm}1.26\;mg/L\;and\;30.70{\pm}0.28\;mg/L\;respectively)$ over the control cultures $(4.70{\pm}0.27 mg/L)$. $5\%$ v/v cell extracts of Fusarium solani has shown moderate increase in the content of azadirachtin $(5.71{\pm}0.34\;mg/L)$ when compared to control cultures $(2.40{\pm}0.56\;mg/L)$. The addition of methyl jasmonate $(500\;{\mu}M/L)$ on day 3 has shown $\~4$ fold improvement in bioproduction of azadirachtln $(6.92{\pm}0.11\;mg/L)$ when compared to control cultures $(1.63{\pm}0.02\;mg/L)$. There was no significant effect of the studied growth retardants and permeabilizing agents on bioproduction of azadirachtin. Cells are cultivated in large volumes using the effective precursors.

• The Korean Journal of Pesticide Science
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• v.19 no.2
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• pp.88-92
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• 2015

This study investigated the thermal stabilities of the four limonoidal active compounds like azadirachtin A, azadirachtin B, deacetylsalannin and salannin and of the two alkaloidal active compounds like matrine and oxymatrine in commercial biopesticides at incubated conditions ($30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $45^{\circ}C$, and $54^{\circ}C$). Half-lives of the limonoidal compounds in biopesticide ranged from 25.6 to 220 days. And the total limonoid contents changed over 15% after 14 days at all the conditions in the tested biopesticide containing neem extract. On the contrary, half-lives of the alkaloidal compounds in biopesticide showed the ranges from 231 to 346 days. And total alkaloidal compounds in all the tested biopesticides containing Sophora flavescens extracts decomposed below 15% after 14 days incubation at $54^{\circ}C$.

• Korean Journal of Organic Agriculture
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• v.22 no.3
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• pp.435-444
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• 2014

This experiment was carried out to develop an environmental friendly agriculture material (EFAM) using neem extract i.e. azadirachtin (0.5%), especially against Bemisia tabaci (Gennadius) in eggplant. Neem product was found to be very effective against both nymph and adult of B. tabaci. Yield and economic value of eggplant treated with neem extract were higher than other EFAMs used. Effect of different concentration of Neem product was also investigated on population density of B. tabaci. The growth and yield of eggplants were highest in plants treated with 500 times concentrated solution of neem product. Economic analysis showed that the income increased by 13,545 thousand won/10a from the plots treated with 500 times concentrated solution of neem product over control. The results suggested that 500 times concentrated solution of EFAM (azadirachtin 0.5%) could be a good controlling agent of B. tabaci in eggplants.

• Korean journal of applied entomology
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• v.60 no.4
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• pp.463-471
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• 2021

As a tropical plant, the neem tree (Azadirachta indica) has been used for a long time for disease and pest control and medical purposes. In this paper, we reviewed for the active ingredient of neem, the mode of action of azadirachtin in terms of insect growth regulation, repellent, feeding inhibition and oviposition against plant pests. And also we reviewed the current status of practical use in Korea. Among 57 products of neem-based eco-friendly organic agricultural materials distributed in Korea, seven products were certified for their efficacy. The average azadirachtin content of the seven products was 0.38%, which was 5.5 times less than the average content of 2.1% distributed worldwide. In the control effect on neem products in Korea, it showed some variation on aphids, but most showed a control effect of more than 90%. The treatment effects of Lycorma delicatula nymphs were 73-77%, and the control effects for thrips were obtained in the range of 50-72% mortality. The mortality effects against bug and moth species were generally low. It is expected that this review would provide important information necessary for the understanding of distributed neem products and the interpretation of experimental data.

• 한국약용작물학회:학술대회논문집
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• 2009.05a
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• pp.375-376
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• 2009