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Isolated culture method of ginseng stem cells

A technology for separating and cultivating stem cells, which is applied in cell dissociation methods, plant cells, and cultivation processes, etc. It can solve the problems of weak division ability, limited division ability, and weak resistance to stress, etc., so as to increase the content of active ingredients and reduce pollution Risk, the effect of shortening the induction time

Active Publication Date: 2021-07-27
SHANDONG ANRAN NANOMETRE IND DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this kind of cell essence comes from differentiated somatic cells, so the ability to divide is limited and the ability to resist stress is not strong
In industrial production, cell line degeneration is prone to occur, and the ability to divide is weak
In addition, the culture period is long, and the content of active ingredients in the obtained stem cells also needs to be improved.

Method used

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  • Isolated culture method of ginseng stem cells
  • Isolated culture method of ginseng stem cells
  • Isolated culture method of ginseng stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] In this example, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0065] (1) Induction of adventitious roots

[0066] After 20-year-old wild ginseng, remove the reed head, stalk, and main root after cleaning, disinfection and sterilization, cut into thin slices with a width of 0.6cm, a length of 0.7cm and a thickness of 0.3mm, and inoculate them into induction medium, at 22±1°C Adventitious roots of wild ginseng were induced by dark culture for 4 to 5 weeks under conditions; wherein, the induction medium included 4mg / L naphthaleneacetic acid, 0.6mg / L gibberellin, 0.4mg / L kinetin, 0.1g / L citric acid, 0.05g / L L ascorbic acid, 30g / L sucrose, 3g / L Phytogel, 1.55g / L B5 medium and 1.21g / L WPM medium, pH value is 5.8.

[0067] (2) Subculture of adventitious roots

[0068] Inoculate the adventitious roots of wild ginseng obtained in step (1) into the same induction medium as ...

Embodiment 2

[0073] In this example, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0074] (1) Induction of adventitious roots

[0075] After cleaning, disinfecting and degerming the reed heads of 6-year-old garden ginseng, cut them into thin slices with a width of 0.5 cm, a length of 0.6 cm and a thickness of 0.3 mm, inoculate them into induction medium, and culture them in dark at 22±1°C for 4-5 days. Zhou induced adventitious roots; wherein, the induction medium included 6mg / L naphthaleneacetic acid, 0.2mg / L gibberellin, 0.4mg / L kinetin, 1.2g / L citric acid, 0.1g / L ascorbic acid, 20g / L sucrose, 5g / L Phytogel, 4g / L B5 medium and 1.8g / L WPM medium, the pH value is 5.6.

[0076] (2) Subculture of adventitious roots

[0077] Inoculate the adventitious roots obtained in step (1) into the same induction medium as in step (1), and culture in dark under the same conditions for 4 to 5 weeks; ...

Embodiment 3

[0082] In this example, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0083] (1) Induction of adventitious roots

[0084] After cleaning, disinfecting and sterilizing the horns of 10-year-old understory ginseng, cut them into thin slices with a width of 0.7 cm, a length of 0.7 cm, and a thickness of 0.5 mm, inoculate them into induction medium, and culture them in dark at 22±1°C for 4~ Adventitious roots were induced in 5 weeks; wherein, the induction medium included 5mg / L naphthaleneacetic acid, 1mg / L gibberellin, 0.1mg / L kinetin, 0.75g / L citric acid, 0.03g / L ascorbic acid, 40g / L sucrose, 4g / L Phytogel, 2g / L B5 medium and 1g / L WPM medium, pH 6.0.

[0085] (2) Subculture of adventitious roots

[0086] Inoculate the adventitious roots obtained in step (1) into the same induction medium as in step (1), and culture in dark under the same conditions for 4 to 5 weeks;

[0087...

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Abstract

The invention discloses an isolated culture method of ginseng stem cells. The isolated culture method comprises the following steps: (1) preparing ginseng adventitious roots by adopting a one-step method; (2) taking the root tip part of the ginseng adventitious root prepared in the step (1), and dissecting and separating to obtain a stem cell region; (3) inoculating the stem cells obtained by separation in the step (2) into a stem cell induction medium for dark culture to obtain a stem cell mass, and then transferring the stem cell mass into a subculture medium for dark culture; and (4) inoculating the stem cells subjected to subculture in the step (3) into a stem cell liquid culture medium, and performing dark culture to obtain the ginseng stem cells. By adopting the liquid culture medium and the culture method disclosed by the invention, adventitious roots can be directly generated through induction of each part of mature ginseng, the content of ginsenoside in the adventitious roots is increased while the induction steps are simplified and the induction time is shortened, and finally ginseng stem cells with good indexes are obtained.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for separating and culturing ginseng stem cells. Background technique [0002] Ginseng (Panax ginseng C.A.Mey.) is a plant of the genus Panax of Araliaceae, distributed in China, Japan, and Korea. Its rhizome is a valuable Chinese medicinal material, known as the "King of Herbs". Ginseng is sweet, slightly bitter, and slightly warm. It has the functions of invigorating vital energy, restoring pulse and strengthening the spleen, nourishing the spleen and lung, promoting body fluid and nourishing blood, calming the nerves and improving intelligence, etc. Wound thirst, palpitations, insomnia, etc. Ginsenoside is the main active ingredient of ginseng, which has the effects of anti-fatigue, anti-aging, regulating the central nervous system, improving immunity, improving cardiovascular and cerebrovascular insufficiency, and inhibiting the production o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04C12N2509/00C12N2500/34C12N2500/40C12N2500/30
Inventor 刘冰高秀君闫培生张明臣
Owner SHANDONG ANRAN NANOMETRE IND DEV CO LTD
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