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Tissue culture method for promoting liquorice adventive root glycyrrhizic acid accumulation by means of fungal elicitors

A fungal elicitor and elicitor technology, applied in cell culture medium, plant cells, etc., to achieve low production cost, simple operation, and increase the content of glycyrrhizic acid compounds

Inactive Publication Date: 2016-07-06
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no report on the use of Pichia pastoris to promote the accumulation of glycyrrhizic acid in adventitious roots of Glycyrrhizae

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The tissue culture method using fungal elicitor to promote the accumulation of glycyrrhizic acid in the adventitious roots of licorice includes the following steps:

[0023] (1) Inoculate licorice roots in a solid medium to induce the formation of licorice adventitious roots, inoculate licorice adventitious roots in a liquid medium, and culture for 35 days;

[0024] (2) Pichia pastoris was fermented and cultured in PDA medium at room temperature for 5 days, centrifuged to obtain cells, dried cells at 60℃, ground into powder and added water to make a suspension with a concentration of 1mg / ml, sterilized , Which is the elicitor suspension;

[0025] (3) Add the elicitor suspension to the liquid medium of the adventitious roots of licorice that has been cultured for 35 days in step (1), so that the final concentration of the elicitor is 200 mg / L, continue to cultivate for 5 days, and harvest.

Embodiment 2

[0027] The tissue culture method of using fungal elicitor to promote the accumulation of glycyrrhizic acid in the adventitious roots of licorice includes the following steps:

[0028] (1) Inoculate the stems of licorice in a solid medium to induce the formation of adventitious roots of licorice, inoculate the adventitious roots of licorice in a liquid medium, and cultivate for 33 days;

[0029] (2) Pichia pastoris was fermented and cultured in PDA medium at room temperature for 6 days, centrifuged to obtain the bacteria, dried at 58℃, ground into powder, added water to make a suspension with a concentration of 1mg / ml, sterilized , Which is the elicitor suspension;

[0030] (3) Add the elicitor suspension to the liquid medium of the adventitious roots of licorice that has been cultured for 33 days in step (1), so that the final concentration of the elicitor is 300 mg / L, continue to cultivate for 6 days, and harvest.

Embodiment 3

[0032] The tissue culture method of using fungal elicitor to promote the accumulation of glycyrrhizic acid in licorice adventitious roots includes the following steps:

[0033] (1) Inoculate the leaves of licorice in a solid medium to induce the formation of adventitious roots of licorice, inoculate the adventitious roots of licorice in a liquid medium, and cultivate for 37 days;

[0034] (2) Pichia pastoris was fermented and cultured in PDA medium at room temperature for 4 days, centrifuged to obtain the bacterial cells, dried at 62℃, ground into powder, added water to make a suspension with a concentration of 1mg / ml, sterilized , Which is the elicitor suspension;

[0035] (3) Add the elicitor suspension to the liquid medium of the adventitious roots of licorice that has been cultivated for 37 days in step (1), so that the final concentration of the elicitor is 100 mg / L, continue to cultivate for 4 days, and harvest.

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Abstract

The invention discloses a tissue culture method for promoting glycyrrhizic acid accumulation in licorice adventitious roots by using fungal elicitors, comprising the following steps: (1) inoculating licorice roots, stems or leaves in a solid medium to induce the formation of licorice adventitious roots, and The adventitious roots of licorice are inoculated in liquid medium and cultured; (2) Pichia pastoris is fermented and cultured at room temperature for 4-6 days in PDA medium, centrifuged to obtain the bacteria, dried, ground into powder and added to water to make a suspension (3) Add the elicitor suspension to the liquid medium of the adventitious root of Glycyrrhizae that has been cultivated for 33-37 days in step (1), so that the final concentration of the elicitor is 100-300mg / L, continue to cultivate for 4‑6 days, and harvest. The method of the invention has the advantages of simple operation, can significantly increase the content of glycyrrhizic acid compound in the adventitious root of licorice root, has short growth period, low production cost, and is convenient for large-scale industrialized production of glycyrrhizic acid.

Description

Technical field [0001] The invention relates to a tissue culture method for promoting the accumulation of glycyrrhizic acid compounds in the adventitious roots of licorice by using fungal elicitors, and belongs to the field of tissue culture of Chinese medicine licorice. Background technique [0002] Glycyrrhizic acid (Glycyrrhizic acid) is a type of E-type saponins and is the most abundant saponins in licorice. As the main active ingredient in licorice, glycyrrhizic acid mainly has the following uses: [0003] (1) Pharmaceutical industry: Glycyrrhizic acid has the functions of anti-inflammation and liver protection, anti-virus, anti-tumor, interferon inducer, and regulating immune function. It is clinically used in the prevention and treatment of chronic hepatitis, bronchitis, AIDS and various cancers. (2) Food and cigarette industry: Glycyrrhizic acid is a health-care sweetener with high sweetness and low calorific value. Its sweetness is about 250 times that of sucrose. It can ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04C12N5/0025
Inventor 高文远王娟李建丽李静李金鑫刘书杰
Owner TIANJIN UNIV
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