Method for producing podophyllotoxin induced from sino podophyllum hexandrum callus

A technology of callus induction and podophyllotoxin, which is applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problem of unreported callus-induced podophyllotoxin, low content, Problems such as difficulty in callus culture

Inactive Publication Date: 2008-02-06
GANSU ACAD OF SCI INST OF BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Foreign countries have obtained podophyllotoxin from the callus and suspension culture system of Podophyllum peltatum and Podophyllum hexandrum Royle, but the content is very low, and it has been proved that the callus of plants containing podophyllum toxin Tissue culture is very difficult; it is only reported in China that the content of podophyllotoxin obtained from cultured plants of the genus Illicium is half of that of the original plant, but there is no report on callus induction of this plant
In China, there are some reports on the distribution pattern, seed characteristics, endophytic fungi that produce podophyllotoxin, the extraction method of podophyllotoxin and the medicinal use of podophyllotoxin, etc. The tissue culture, callus induction and podophyllotoxin production of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1. Put the selected and plump peach seven seeds produced in Longnan, Gansu Province into a beaker, slowly add concentrated sulfuric acid, stir continuously with a glass rod for 10 minutes, rinse with tap water until the sulfuric acid is completely removed. Replace with 300mg / L gibberellin solution for 48 hours. The treated seeds are sterilized with 0.1% mercury liter for 5-6 minutes under aseptic conditions, rinsed with sterile water for several times, and blotted dry with sterile filter paper. Embryos were taken out and inoculated into modified MS medium (1 / 2 MS inorganic components, no hormones), and cultured in a light incubator. Other conditions are: agar 0.75%, sucrose 30g / L, pH adjustment is 5.8; Illumination (500μmol.m -2 .s -1 ) for 10 hours at a temperature of 25±1°C. The embryo begins to swell in about 3 days, two cotyledons unfold at 6 days, the cotyledons are green at 10 days, the radicle and hypocotyl elongate obviously at 20 days, and grow into complete...

Embodiment 2

[0020] 1. select four kinds of explants from the aseptic seedling that step 1-3 obtains among the embodiment 1: the aseptic seedling that grows about 40 days ① hypocotyl ② radicle ③ cotyledon and ④ true leaf petiole of about 60 days, cut into small pieces of about 0.3 cm or 0.3 cm square, connected to B 5 The induction medium (explant ①, ② and ③) is selected from B 5 Add 2,4-D 1mg / L, 6-BA 3mg / L and NAA 1mg / L; ④Choose B 5 Additional 2,4-D 5mg / L, 6-BA 1mg / L and NAA 0.5mg / L). Other conditions are: agar 0.75%, sucrose 20g / L, pH adjustment is 5.8; Illumination (500μmol.m -2 .s -1 ) for 10 hours at a temperature of 23±1°C. After about 40-50 days of culture, the four different explants all induced callus.

[0021] 2. Screen callus with loose texture and fast proliferation and subculture continuously on different subculture media respectively: callus from explant ①, ② and ③) source selection B 5 Add 2,4-D 0.2mg / L, 6-BA 1mg / L and NAA 1mg / L, callus from true leaf petiole selection...

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Abstract

The present invention discloses a method for inducing callus of podophyllum emodi to produce podophyllotoxin. Said method includes the following steps: using wild podophyllum emodi seed as original raw material to obtain sterile seedling of podophyllum emodi, multiplication and rooting, using different regions of said sterile seedling as explant, further inducing callus, multigeneration screening and successive transfer culture so as to create the callus system suitable for making large-scal cell culture, in said culture the target metabolic product-podophyllotoxin can be contained.

Description

technical field [0001] The invention relates to a method for cultivating aseptic seedlings and inducing callus to produce podophyllotoxin of P. chinensis. Background technique [0002] At present, due to the declining success rate of chemically synthesized new drugs, increasing costs, prolonging the cycle, and relatively large side effects, it has become a worldwide trend to extract secondary metabolites from natural plants for direct medicinal use, or as precursors for chemically semi-synthesized new drugs. . A quarter of the prescriptions in the United States contain medicines derived from plants, and there are still more than 50 important medicines that can only be extracted from plants. 80% of my country's traditional Chinese medicine is wild resources. Human beings' great demand for wild plant resources has caused serious damage to the ecological environment and the endangered status of many wild plants. Relevant studies at home and abroad have proved that the use of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C07D493/04
Inventor 杨晖王治业陆栋魏甲乾龚伟中周剑平孙智敏张文齐
Owner GANSU ACAD OF SCI INST OF BIOLOGY
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