Azadirachta indica cell suspension culture medium and cell suspension culture method

A cell suspension and culture method technology, which is applied in the field of neem cell suspension medium and cell suspension culture, can solve the problems of slow biomass accumulation and low content of azadirachtin, and achieves the goal of promoting cell growth and metabolism, increasing content and reducing browning. The effect of conversion rate

Active Publication Date: 2019-06-14
无锡吉成生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention is aimed at the defect that prior art exists, provides a kind of neem cell suspension culture medium and cell suspension cult

Method used

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  • Azadirachta indica cell suspension culture medium and cell suspension culture method
  • Azadirachta indica cell suspension culture medium and cell suspension culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0028] A kind of neem cell suspension culture medium, the formula of this culture medium is as follows:

[0029] KNO 3 2250mg / L, (NH 4 ) SO 4 500mg / L, MgSO 4 ·7H 2 O 235mg / L, NaH 2 PO 4 246mg / L, CaCl 2 2H 2 O 534mg / L, MnSO 4 4H 2 O 10mg / L, H 3 BO 3 3.4mg / L, ZnSO 4 ·7H 2 O 3.8mg / L, KI 0.54mg / L, CuSO 4 ·5H 2 O 0.025mg / L, CoCl 2 ·6H 2 O 0.025mg / L, Na 2 MoO 4 2H 2 O 0.23mg / L, FeSO 4 ·7H 2 O23.5mg / L, Na 2 -EDTA 32.5mg / L, inositol 100mg / L, glycine 4mg / L, thiamine hydrochloride 1.1mg / L, pyridoxine hydrochloride 0.6mg / L, sodium vanadate 1.6mg / L, L-cysteine 35mg / L, tyrosine 20mg / L, nicotinamide 0.6mg / L, dithiothreitol 1.6mg / L, β-D-galactose 2.75mg / L, cyclohexyl phosphate 9.7mg / L, Diethyl hexanoate 0.3mg / L, 6-BA 0.5mg / L, ecdysone 2.6mg / L, sodium nitroprusside 0.2mmol / L, dioctyl sodium sulfosuccinate 0.13g / L, Polygonum japonica juice 16mL / L, Rehmannia rhizome juice 15mL / L, sucrose 19g / L, glucose 13g / L, pH value 5.6; Among them, the fresh stems and leaves of ...

Embodiment 2

[0037] The optimization test of embodiment 2 culture conditions

[0038] 2.1 Effect of pH value of cell suspension medium on the growth of neem suspension cells and the content of azadirachtin

[0039] Adjust the pH value of the cell suspension medium to 4.8, 5.2, 5.6, 6.0, 6.4 respectively, cultivate according to the neem suspension cell culture method in Example 1, collect the suspension cells, and measure the cell biomass and neem at different pH values. neem content, the results were as figure 1 shown. From figure 1 It can be seen that with the increase of the pH value of the medium, the cell biomass and azadirachtin content showed a trend of first increasing and then decreasing. When the medium pH value was 5.6, the neem cell biomass and azadirachtin content were both Reached the maximum value, so 5.6 is the optimal medium pH value.

[0040] 2.2 The effect of inoculum size on the growth of neem suspension cells and the content of azadirachtin

[0041] The callus is...

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Abstract

The invention provides an azadirachta indica cell suspension culture medium and a cell suspension culture method. According to the azadirachta indica cell suspension culture medium, based on a basic culture medium, component matching is optimized, and a certain amount of sodium vanadate, L-cysteine, tyrosine, nicotinamide, dithiothreitol, beta-D-galactose, cyclohexanol phosphate, diethylaminoethylhexanoate, 6-BA, ecdysone, sodium nitroprusside, docusate sodium, flaccid knotweed herb juice and henry munronia herb juice is added; the cell suspension culture method comprises the following stepsof 1) obtaining of sterile test-tube plantlets; 2) induction and subculture for calluses; 3) cell suspension culture. By adopting the culture medium and the culture method, not only can the stable azadirachta indica cell suspension culture system be obtained, but also the biomass of suspension cells and the content of azadirachtin in the suspension cells can be significantly improved, the technical support is provided for azadirachtin production through large-scale azadirachta indica cell culture, and important application value is achieved.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a neem cell suspension medium and a cell suspension culture method. Background technique [0002] Neem (scientific name: Azadirachta Indica A.Juss.) is a plant of the genus Neem of the family Meliaceae. It is mainly distributed in the tropical regions of Southeast Asia. It has been introduced and cultivated in Guangdong, Hainan, Sichuan and Yunnan in my country. And fast-growing tropical or subtropical tree species. Neem is recognized as an ideal insecticidal plant in the world. With its unique insecticidal effect and high-efficiency insecticidal, antifeedant, repellent, poisonous and fertility-inhibiting activities, it ranks first in the botanical pesticide resources. Active substances are contained in neem fruit, seeds, seed cores, branches, leaves, bark and sap, but especially in seeds. So far, more than 100 compounds have been found in neem, at least 70 com...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 杨友霞
Owner 无锡吉成生物科技有限公司
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