Saussurea involucrata cell lines

A technology of cell line and snow lotus, which is applied in the direction of plant cells, digestive system, plant raw materials, etc., can solve the problems of reduced active ingredients and instability, and achieve the effects of shortened production cycle, high stability and vigorous growth

Active Publication Date: 2009-01-07
DALIAN PRACTICAL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no report about further increasing the flavonoid content
In addition, the snow lotus cell line is unstable during the scale-up process, and the content of active ingredients will decrease when the scale-up culture reaches a certain scale. At present, there is no effective means to solve the above problems

Method used

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  • Saussurea involucrata cell lines
  • Saussurea involucrata cell lines
  • Saussurea involucrata cell lines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Obtaining of Xinjiang Saussurea sauvignon Violet Cell Line

[0028] Take natural Xinjiang snow lotus seeds, first soak them in 75% alcohol for 10-30s, then sterilize them with 1% mercuric chloride solution for 8-12mins, then rinse them repeatedly with sterile water for 4-5 times, and finally put them on sterile filter paper for absorption. Dried seed surface moisture, inoculated on the solid medium of 1 / 2MS (recipe sees attached table 1) with 2.5mg / L GA (gibberellin), 30g / L sucrose, 0.6g / L agar with sterile tweezers, Cultivate under light at 25°C, and the seeds can germinate and grow sterile seedlings in about 10 days.

[0029]Take aseptic seedling germs as explants, under aseptic conditions, use sterile surgical scissors and sterile tweezers to cut the sterile seedling germs into 2-5mm long segments, and inoculate them in MS (see attached table 1 for the formula) Add 0.4mg / L 6-BA (6-benzylpurine), 3mg / L NAA (naphthalene acetic acid), 0.6g / L agar and 30g / L su...

Embodiment 2

[0035] Embodiment 2 liquid-solid alternate production method

[0036] 1. Liquid shaker culture and seed production

[0037] 1.1 Selection

[0038] The culture of the cell line Sau-1 of the present invention, which has been cultured for 10-14 days, has good growth, bright color, purple-red, uniform particles, uniform texture and is easy to separate, is selected as the seed.

[0039] 1.2 Culture medium preparation

[0040] Use 500ml culture bottles, each bottle is filled with 120-150ml culture medium, the composition of the culture medium is MS (see attached table 1 for the formula) and add 0.4mg / L 6-BA (6-benzylpurine), 3mg / L NAA (naphthalene acetic acid) and 30g / L sucrose, the pH value is 5.5; high temperature sterilization: 121°C, 20min.

[0041] 1.3 Inoculation

[0042] Under sterile conditions, the culture was crushed with tweezers and inoculated into a 500-ml culture bottle filled with 120-150 ml of culture solution, with an inoculum volume of 5% (W / V, g.fw / ml).

[00...

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Abstract

The invention provides a purple cell system of a Saussurea Involucrata Sau-1 CGMCC No.2506 which stably products flavone with high yield. The content of a flavone compound can achieve 8 to 12 percent of the dry weight of a culture which is 7 to 10 times of a natural Involucrata. The invention also provides a culture technology of the cell system and a method for fast producing the culture of the cell system in a large scale. The method adopts the combination of liquid-solid culture, manufactures the seeds by liquid and obtains the culture by solid culture, selects the seeds with better comprehensive situations for liquid culture and repeats in the way. The method can be used for large scale production. Compared with a wild type, the production period is greatly shortened; besides, the quality is stable and the yield is high. Besides, the method does not occupy the farming land, does not damage the wild resource of the Involucrata., protects the environment and solves the problem of lack of the source of Saussurea Involucrata.

Description

technical field [0001] The invention relates to a snow lotus cell line, and also relates to a preparation method of the cell line and a large-scale production method of its culture. Background technique [0002] Snow lotus is a folk medicinal plant in high mountain areas. It is used for dispelling cold and dehumidification, promoting blood circulation and stimulating menstruation, anti-inflammatory and analgesic, etc. It is mostly used in folk for rheumatoid arthritis, cold pain in women's lower abdomen, amenorrhea, retained placenta, impermeable numbness, and cold lung Treatment of cough, impotence, and altitude sickness. In recent years, Snow Lotus, as a national medicine, has attracted much attention for its anti-inflammatory and analgesic effects, anti-early pregnancy, anti-aging and inhibition of cancer cell proliferation. But my country's snow lotus is mainly distributed in the plateau cold zone above 4000m, such as Xinjiang, Gansu, Sichuan, Yunnan and Tibet. The cli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04A23L1/29A61K36/285A61P1/14C12R1/91A23L33/00
Inventor 刘汉石
Owner DALIAN PRACTICAL BIOTECH
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