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Method for producing saussurea medusa maxim culture rich in flavone and polysaccharide

A technology of Saussurea oleifera culture and production method, which is applied in the field of production of Saussurea oleifera secondary metabolites, can solve the problems of unexplained influence of flavonoids on cell aggregation growth, poor genetic stability of cells, and decrease of flavonoid content, etc., and achieve good product quality controllability, The effect of short culture period, stable yield and content

Inactive Publication Date: 2014-01-01
GUANGZHOU ENDAO ECOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is an effective way to solve the problem of snow lotus resources through artificial cultivation, but there is no literature report on the use of cells to cultivate high-yield snow lotus polysaccharides.
[0004] Chinese invention patent application (publication number CN1340611A, publication date March 20, 2002, application number 00123740.3) discloses a high-yield flavonoid cell line of Saussurea jellyfish, using the cell line and medium provided by the invention, the cell growth rate is 35 ~40g dry weight / liter / month, the content of flavonoids is 6~7% of culture dry weight, although have better cell growth characteristics and higher ability of biosynthetic flavones, but still have the following deficiencies:( 1) The flavonoid content is still low; (2) The flavonoid content may decrease if the cell aggregate culture size is larger than Smm; (3) The genetic stability of the cells is poor
[0005] Chinese invention patent application (publication number CN1424395A, publication date: June 18, 2003, application number 02156868.5) discloses a method for establishing a stable and high-yield secondary metabolite flavonoid Saussurea chinensis cell line, which utilizes 60Co-γ-ray irradiation, ultraviolet The original snow lotus cell line was mutagenized by light irradiation and other means to increase the content of flavonoids in the snow lotus cell, and a stable and high-yielding snow lotus cell line was obtained, which can be continuously passaged for 50 generations. weight) / L and about 12%. This cell line has strong adaptability and can grow in an environment of 18°C ​​to 30°C. The optimum culture temperature range is 20°C to 25°C. Synthetic Effects

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0026] The present embodiment relates to a kind of production method of Saussurea edulis culture rich in flavonoids and polysaccharides, comprising the following steps:

[0027] Step (1): Snow lotus seeds are germinated to obtain snow lotus plants, and the snow lotus plants are used as explants, rinsed with tap water first, then soaked in a fungicide to sterilize, then rinsed with sterile water 5 times for 2 minutes each time, and then inoculated in Culture on 1 / 2MS medium containing 2,4-dichlorophenoxyacetic acid under light at 24°C. After 12 days of culture, callus tissue can be formed at the incision site of the explant. Subculture, subculture once every 15 days, select the cell line of the light red callus as the cell line with high yield of Saussurea polysaccharide;

[0028] Step (2), culturing the above-mentioned cell lines in a basic medium supplemented with colchicine to obtain a polyploid cell line;

[0029] Step (3), inoculating the above polyploid cell line of Saus...

Embodiment 2

[0031] The present embodiment relates to a kind of production method of Saussurea edulis culture rich in flavonoids and polysaccharides, comprising the following steps:

[0032] Step (1), snow lotus seeds are germinated to obtain snow lotus plants, and the snow lotus plants are used as explants, first washed with tap water, then soaked in a fungicide to sterilize, and then rinsed with sterile water 6 times for 3 minutes each time, and then inoculated in Culture on 1 / 2MS medium containing 6-benzyl adenine under light at 25°C. After 13 days of culture, callus tissue can be formed at the incision site of the explant, and the callus tissue can be subcultured on the basic medium , once every 14 days, the cell line of the light red callus was selected as the cell line with high yield of Saussurea polysaccharide;

[0033] Step (2), culturing the above-mentioned cell lines in a basic medium supplemented with colchicine to obtain a polyploid cell line;

[0034] Step (3), inoculating t...

Embodiment 3

[0036] The present embodiment relates to a kind of production method of Saussurea edulis culture rich in flavonoids and polysaccharides, comprising the following steps:

[0037] Step (1): Snow lotus seeds are germinated to obtain snow lotus plants, and the snow lotus plants are used as explants, rinsed with tap water first, then soaked in a fungicide to sterilize, then rinsed with sterile water 5 times for 2 minutes each time, and then inoculated in Culture in the dark at 26°C on 1 / 2MS medium containing naphthaleneacetic acid. After 14 days of culture, callus tissue can be formed at the incision site of the explant, and the callus tissue is subcultured on the basic medium. For one generation, the cell line of the light red callus is selected as the cell line of high yield Saussurea polysaccharide;

[0038] Step (2), culturing the above-mentioned cell lines in a basic medium supplemented with colchicine to obtain a polyploid cell line;

[0039] Step (3), inoculating the above ...

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Abstract

The invention discloses a method for producing a saussurea medusa maxim culture rich in flavone and polysaccharide. The method includes the following steps: washing a saussurea medusa maxim explant through tap water; then steeping and sterilizing the explant through a sterilizing agent; then washing the explant through sterile water; then inoculating the sterilized explant on a 1 / 2MS culture medium including plant hormones and cultivating for 12-15 d at the temperature of 24-26 DEG C, namely, forming a callus at a cut part of the explant, subculturing the callus on a minimal medium, and subculturing once every 12-15 days, and selecting a cell line of a light red callus as a cell line of high-yield saussurea medusa maxim polysaccharide; adding the cell line to a minimal medium of colchicine for cultivation, so as to obtain a polyploidy cell line; inoculating the polyploidy cell line of saussurea medusa maxim to a synthetic medium for cultivation, so as to obtain cells rich in secondary metabolism products of saussurea medusa maxim. Through the adoption of the method, industrial production of secondary metabolism products of saussurea medusa maxim, containing the flavone and the polysaccharide can be stably realized, the production cost is low, the cultivation period is short, and the method is free from the influence of natural conditions.

Description

technical field [0001] The invention relates to a method for producing secondary metabolites of snow lotus, in particular to a method for producing a culture of snow lotus rich in flavonoids and polysaccharides. Background technique [0002] Snow lotus (Saussurea medusa Maxim) is a precious herbal medicine in my country. It is produced in the snowy areas of high-altitude mountainous areas in Gansu, Qinghai, Tibet, Xinjiang and other places in China. It is used in the folk for the treatment of rheumatoid arthritis, impotence, irregular menstruation in women, and maladaptive symptoms of high mountains. Pharmacological studies have proved that Saussurea sauraceae has anti-rheumatism and analgesia, cardiac strengthening, termination of pregnancy, scavenging free radicals and anti-fatigue, antispasmodic, antihypertensive and anti-asthma, inhibition of virus replication, inhibition of chromosome damage, promotion of radiation damage repair, promotion of wound healing and anti-infla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N15/01A01H4/00C12R1/91
Inventor 陈泉
Owner GUANGZHOU ENDAO ECOLOGICAL SCI & TECH
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