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Sinkiang alkanna tinctoria cell lines and process for producing sinkiang alkanna tinctoria alkannin by cell culture

A Xinjiang comfrey, cell line technology, applied in plant cells, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve the problems of high production conditions, high production costs, complex processes, etc. The effect of shortening the production cycle and protecting the ecological environment

Inactive Publication Date: 2008-10-22
DALIAN PRACTICAL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on Xinjiang Zicao is mainly focused on the former, but because of the disadvantages of high production conditions, complicated process, long process and high production cost, the popularization and application of this technology has been greatly restricted, and further research is needed

Method used

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  • Sinkiang alkanna tinctoria cell lines and process for producing sinkiang alkanna tinctoria alkannin by cell culture
  • Sinkiang alkanna tinctoria cell lines and process for producing sinkiang alkanna tinctoria alkannin by cell culture
  • Sinkiang alkanna tinctoria cell lines and process for producing sinkiang alkanna tinctoria alkannin by cell culture

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Experimental program
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Effect test

Embodiment 1

[0028] The acquisition of embodiment 1 Xinjiang Zicao cell line

[0029] Take natural Xinjiang comfrey seeds, first soak them in 75% alcohol for 10-30s, then sterilize them with 1% mercuric chloride solution for 8-12 minutes, then rinse them repeatedly with sterile water for 4-5 times, and finally put them on sterile filter paper Blot the water on the surface of the seeds, use sterile tweezers to inoculate on the solid medium added to basic MS (recipe in Attached Table 1), 10-30g / L sucrose, 0.6g / L agar, and culture at 23-25°C without light. Seeds can germinate and grow aseptic seedlings in about 10 days.

[0030] Take sterile embryos as explants, under aseptic conditions, use sterile surgical scissors and sterile tweezers to cut the sterile seedling roots, stems, leaves, cotyledons, and embryos into 2-5mm long segments, and inoculate them in N6 ( See attached table 2 for the formula, add 0.2mg / L-0.5mg / L IAA, 1mg / L-5mg / L KT, 0.6g / L agar and 30g / L sucrose on solid medium, culture...

Embodiment 2

[0033] Embodiment 2 liquid-solid alternate production method

[0034] 1. Liquid shaker culture and seed production

[0035] 1.1 Selection

[0036] Select and cultivate for 20-25 days, and the cell line culture of the present invention that grows well, brightly colored, purple-red, uniform in particle size, uniform in texture, and easy to separate is used as the seed.

[0037] 1.2 Culture medium preparation

[0038] Use 500ml culture bottles, each bottle contains 120-150ml culture solution, the composition of the culture solution is N6 (refer to attached table 2 for formula), add 0.2mg / L-0.5mg / L IAA, 1mg / L-5mg / L KT, and 30g / L L sucrose, pH value 5.2-5.8; high temperature sterilization: 121°C, 20min.

[0039] 1.3 Inoculation

[0040] Under sterile conditions, crush the culture with tweezers and inoculate it into a 500-ml culture bottle filled with 120-150 ml of culture solution, with an inoculum size of 3-6% (W / V, g.fw / ml).

[0041] 1.4 Shaker culture

[0042] Culture cond...

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PUM

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Abstract

The invention provides an Arnebia euchroma mauve cell line for stable and high yield of alkannin, wherein, the content of alkannin compounds can reach 6 to 8 percent of the dry weight of culture and is 5 to 9 times of that of wild and natural Arnebia euchroma. The invention also provides a culture technique for the cell line and a method for mass and quick production of the culture of the cell line. The method breaks the routine lithospermum cell two-step culture method, adopts one-step culture method to cultivate the lithospermum cell culture and then realizes production of Arnebia euchroma cells through the method of combination of cell suspension culture and culture of solid culture medium and quick production of effective compositions. The method adopts combination of liquid culture and solid culture, produces seeds through liquid culture, obtains the culture through solid culture and selects and uses reserved seeds with good comprehensive properties for liquid culture, and the steps are repeated. Moreover, the production of the Arnebia euchroma cells by adoption of the method does not occupy arable lands and destroy wild resources of Arnebia euchroma, thereby the environment is protected and the problem of shortage of Arnebia euchroma resources is solved.

Description

technical field [0001] The invention relates to a Xinjiang comfrey cell line, and also relates to a preparation method of the cell line and a production method of its culture. Background technique [0002] Comfrey is a perennial herbaceous medicinal plant whose roots are used as medicine in the Lamiformes Boraginaceae. my country's comfrey resources can be roughly divided into: Xinjiang comfrey (Arnebia euchroma (Royle) Johnst.), hard comfrey (Lithospermum erythrorhizon S.et Z.), Yunnan comfrey (Onosma paniculata Bur.Et Fr.), Inner Mongolia comfrey (Arnebia guttata Bunge.) and Tibetan comfrey (Onosma hookeri Clarke var. Longiflorum Duthie.) and other categories. Among them, Xinjiang comfrey has become the main source of commodity comfrey because of its high pigment content and good pharmacological effects. [0003] Xinjiang comfrey, commonly known as soft comfrey, grows in wild forests or sunny slopes at an altitude of 2500-4200 meters, and is mainly distributed in Xinjiang...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12P7/26A61K8/97A61K36/30A23L1/30A23L33/10
Inventor 刘汉石
Owner DALIAN PRACTICAL BIOTECH
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