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Method for gaining panax japonicus secondary metabolite by using panax japonicus cell culture and biological transformation technique

A technology for biotransformation and metabolites, applied in the field of plant cell culture engineering, can solve the problems of being susceptible to pests and diseases, difficult to artificially cultivate, difficult to reproduce, etc., and achieve the effects of reducing picking and dependence, and excellent and cheap drugs.

Inactive Publication Date: 2008-01-30
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although artificial cultivation has been carried out in Jiangxi, Hunan, Hubei and other places, it has been difficult to realize large-scale artificial cultivation for a long time due to its limited growth, difficulty in reproduction, and susceptibility to pests and diseases.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0022] (1) In the improved RW medium, add α-naphthaleneacetic acid (NAA) 2mg / L and 6-bean aminopurine (6-BA) 4mg / liter, sucrose 20g / L, and then adjust the pH value with acid or alkali Adjust to 5.5-7.0, then add 1.9g / L Gellan Gum, sterilize at 115°C and 0.1MPa pressure for 15 minutes, then cool to make slant induction medium;

[0023] (2) Sterilize the young stems or buds of Panax ginseng as explants through 70% (v / v) ethanol surface sterilization for 50 seconds, then put them into 5% (v / v) sodium hypochlorite solution for sterilization for 20 minutes, and use sterile After rinsing with sterile water for three times, cut off the surface-sterilized shoot tips under aseptic conditions and inoculate them into the above-mentioned slant induction medium. Then transfer to 25°C, 2000 Lux, 12 hours of light conditions for induction culture, and callus will form after 1 month;

[0024] (3) In the modified RW medium, add 1mg / L α-naphthaleneacetic acid (NAA), 2mg / L and 6-bean aminopurin...

example 2

[0032](1) In the improved RW medium, add indole acetic acid (IAA) 2mg / L, kinetin (KT) 4mg / L, sucrose 20g / L, and then adjust the pH value to 5.5-7.0 with acid or alkali, Then add 1.9g / L GellanGum, sterilize at 115°C and 0.1MPa pressure for 15 minutes, then cool to make slant induction medium;

[0033] (2) Sterilize the young stems or buds of Panax ginseng as explants on the surface of 70% ethanol for 30 seconds, then put them into 8% calcium hypochlorite solution for sterilization for 15 minutes, wash them three times with sterile water, and Cut off the surface-sterilized shoot tips under aseptic conditions and inoculate them into the above-mentioned slant induction medium. Then transfer to 25°C, 2000 Lux, 14 hours of light conditions for induction culture, callus formed after 1 month;

[0034] (3) In the improved RW medium, add 1mg / L α-naphthaleneacetic acid (NAA), 2mg / L and kinetin (KT), 30g / L sucrose, 1.9 Gellan Gum, through 115 ℃, 0.1MPa pressure After being sterilized fo...

example 3

[0042] (1) In the improved RW medium, add 2,4-D 1.5mg / L and 6-bean aminopurine (6-BA) 3mg / liter, sucrose 20g / L, and then adjust the pH value with acid or alkali When the temperature is 5.5-7.0, add 6g / L agar, sterilize at 120℃, 0.15MPa pressure for 15 minutes, and cool to make slant induction medium;

[0043] (2) Sterilize the young stems or buds of Panax ginseng as explants on the surface of 70% ethanol for 40 seconds, then sterilize them in 10% sodium hypochlorite solution for 10 minutes, wash them with sterile water three times, and put them under aseptic conditions. Next, cut off the surface-sterilized shoot tip and inoculate it into the above-mentioned slant induction medium. Then transfer to 25°C, 2000 Lux, 16 hours of light conditions for induction culture, callus formed after 1 month;

[0044] (3) In the improved RW medium, add 0.8mg / L 2,4-D, 0.5mg / L and 6-bean aminopurine (6-BA), 30g / L sucrose, 6g / L agar, after 120 ℃, sterilized under 0.15MPa pressure for 15 minutes...

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Abstract

The invention discloses a method by using panax japonicus cell culture and biological transfer technology to obtain secondary metabolites and pertains to plant cell culture engineering field. An improved RW culture medium is added with crane sugar, plant growth hormone and cytokinin and then coagulant; the obtained mixture is made into an induction culture medium after being disinfected in high temperature and then cooled; explants of panax japonicus are inoculated in the induction culture medium under an aseptic condition to be inducted to form callus; after an expansion, a plurality of cells are obtained; the expanded callus in the synthetic medium is inoculated into synthetic culture solution and then panax japonicus saponin crude extracts and byproducts thereof can be obtained through the extraction of cells from the solvent, thereafter the panax japonicus saponin, panax japonicus polysaccharide and other compounds can be obtained through purification. By adopting the method of the invention, secondary metabolites of the panax japonicus can be produced with low cost, short culture cycle, year-round production and no natural environmental limit.

Description

technical field [0001] The invention belongs to the field of plant cell culture engineering, and in particular relates to a method for obtaining secondary metabolites of Panax ginseng by using the cell culture and biotransformation technology of Panax ginseng, especially relates to the method for obtaining secondary metabolites of Panax ginseng by using the large-scale cultivation and biotransformation technology of Panax ginseng cells The method of compounds such as bamboo ginseng saponins. Background technique [0002] Bamboo ginseng (Panax japonicus C.A. Mey), also known as bamboo ginseng or white notoginseng, is a plant of the genus Panax ginseng in the family Araliaceae. Distributed in my country's Jiangxi, Hubei, Guangxi, Sichuan, Guizhou, Yunnan, Tibet and other provinces and regions under alpine bushes, wetlands or beside rocky ditches. It has many different names. The "Dictionary of Chinese Medicine" says: Different books have different titles, such as: soil ginsen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
Inventor 郭志刚孙瑞强
Owner TSINGHUA UNIV
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