Saussurea medusa high yielding flavone cell line and preparation method thereof

The technology of a cell line and snow lotus is applied to the preparation method and medium field of a high-yield flavonoid cell line of jellyfish snow lotus. easy-to-control effects

Inactive Publication Date: 2018-09-07
DALIAN PRACTICAL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, plant cells are used for cultivation, and the flavonoid content of the active ingredient obtained by it is low, usually only about 6-7% of the dry weight. It has always been d...

Method used

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  • Saussurea medusa high yielding flavone cell line and preparation method thereof
  • Saussurea medusa high yielding flavone cell line and preparation method thereof
  • Saussurea medusa high yielding flavone cell line and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] (1) Induction of callus from young embryos of Saussurea tianshanensis

[0040] Disinfect the seeds of Snow Lotus Tianshan with 70% ethanol for 1 min, rinse with sterile water 2-3 times; then sterilize with 2% sodium hypochlorite solution for 60-62 min, rinse with sterile water 4-5 times; under sterile conditions, remove the shell and seed coat, and then connected to the improved MS solid medium (1 / 2), it should be noted that: 1 / 2 refers to the halving of a large number of elements in the medium components, wherein, added in the improved MS medium With 2,4-D (2,4-dichlorophenoxyacetic acid) concentration of 3mg / L, pH value of 5.8, dark culture at 25°C, after 30 days, granules can grow from the edge of the seed hypocotyl and cotyledon cultured in the dark at 25°C, and subcultured every 2 weeks (that is, the medium was replaced every 2 weeks).

[0041] (2) Culture of callus

[0042] The above-mentioned callus was subcultured on the medium of pH 5.8 by adding 0.2 mg / L BA ...

Embodiment 2

[0050] In this example, on the basis of Example 1, the high-yielding red cell line (A line) in Example 1 was used for mutagenesis by 60Coγ-ray irradiation. 5-30Gy of 60Co-γ rays were used to irradiate, the irradiation rate was 0.8Gy / min, and the semi-lethal dose for mutagenic callus was 20Gy. The mutagenized cell lines are then screened by small cell mass, which can refer to the second edition of "Plant Cell Tissue Culture", edited by Liu Qingchang, etc., China Agricultural University Press, 2010.01. 308 page 13.3 Screening of cell lines) to obtain the total flavonoids The higher yielding cell line contained 10.9%.

[0051] Screening of cell aggregates in the liquid state of the A cell line: The above-mentioned high-yield red cell line (A) was cultured in suspension culture conditions (modified medium without agar), and the cell aggregates were passed through a series of different sizes. (1, 2, 3, 4, 5, and 6mm) stainless steel sieves, and weigh the dry weight (Wi) of cell ag...

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Abstract

The invention provides a preparation method and culture medium of a saussurea medusa high yielding flavone cell line. The method comprises the following steps: inducing callus tissue through a saussurea medusa seed; performing solid successive transfer culture on obtained callus tissue; directly selecting a cell line with high total flavone content in the callus tissue subjected to successive transfer culture through a visual method; performing liquid culture, performing solid culture from liquid culture, and continuously performing repeated screening and subculture, so as to obtain a high yielding cell line which is rapid in development and high in total flavone content. The cell line and the culture medium obtained through the method have the advantages that the cell growth rate can reach 45g to 50g dry weight/liter/month when the lighting suspension culture temperature is about 25 DEG C, and the content of flavonoid can reach 12 percent of the dry weight of culture.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method and culture medium of Saussurea jellyfish high-yield flavonoid cell line. Background technique [0002] Jellyfish Snow Lotus (Sausurea medusa Maxim.) is the original plant of Tibetan medicine "Sausurea medusae", distributed in Qinghai, Gansu, Tibet and other places in my country, and is a precious medicinal material in Tibetan medicine. Snow lotus contains flavonoids, alkaloids, lactones, sterols, volatile oils, polysaccharides and other ingredients, and its active ingredients are mainly flavonoids. The pharmacological effects of flavonoids include anti-cancer, anti-tumor, anti-cardiovascular and cerebrovascular diseases, antitussive and expectorant, anti-inflammatory, analgesic effects, immune regulation, estrogen-like effects, antibacterial and antiviral effects, anti-oxidation, anti- Aging effect, anti-radiation. Xuelian injection with flavonoids...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N13/00C12P1/00C12R1/91
CPCC12N1/00C12N5/0025C12N5/04C12N13/00C12P1/00C12R2001/91
Inventor 刘汉石赵德修刘雅萍
Owner DALIAN PRACTICAL BIOTECH
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