Glycyrrhiza glabra callus cell culture method capable of improving content of licoflavone

A technology of callus and licorice glabra, applied in plant cells, fermentation, etc., can solve problems such as inability to meet market demand, and achieve the effects of increasing the content of licorice flavonoids, increasing yield, and increasing reaction rate

Active Publication Date: 2017-02-01
PROYA COSMETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although researchers have made relevant efforts to increase the content of licorice flavonoids in licorice callus cells, in the related results reported, licorice flavonoids only account for about 0.4%~0.7% of the dry weight of callus cells, which is still not satisfactory. Market demand

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A method for culturing callus cells of Glycyrrhiza glabra that can increase the content of licorice flavonoids comprises the following steps:

[0020] 1) Induction and subculture of Glycyrrhiza glabra callus: Select plump Glycyrrhiza glabra seeds and culture them on MS basal medium containing 30g / L sucrose and 7g / L agar to obtain sterile seedlings; take sterile seedling embryos The axes are explants, which are inserted into MS basal medium containing 1.0 mg / L naphthaleneacetic acid, 0.4 mg / L 6-benzylaminopurine, 22 g / L sucrose, and 6 g / L agar, and cultured in the dark for 10 days to induce healing. Injury tissue, then at light intensity 1500 lx The culture was continued for 14 days under the conditions of 12 hours of light every day; then the callus was transferred to the culture medium containing 1.0mg / L naphthaleneacetic acid, 0.4mg / L 6-benzylaminopurine, 0.1mmol / L methionine, 22g / L Sucrose, 6g / L agar MS basal medium, subculture temperature 25 o C, subculture medium...

Embodiment 2

[0024] A method for culturing callus cells of Glycyrrhiza glabra that can increase the content of licorice flavonoids comprises the following steps:

[0025]1) Induction and subculture of Glycyrrhiza glabra callus: Select plump Glycyrrhiza glabra seeds and culture them on MS basal medium containing 30g / L sucrose and 7g / L agar to obtain sterile seedlings; take sterile seedlings Hypocotyls were explants, inserted into MS basal medium containing 1.2mg / L naphthaleneacetic acid, 0.3mg / L 6-benzylaminopurine, 20g / L sucrose, and 8g / L agar, cultured in the dark for 10 days, and induced callus, after which the light intensity was 1500 lx Continue to culture for 14 days under the conditions of 12 hours of light every day; then transfer the callus to the culture medium containing 1.2mg / L naphthaleneacetic acid, 0.3mg / L 6-benzylaminopurine, 0.3mmol / L methionine, 20g / L Sucrose, 8g / L agar MS basal medium, subculture temperature 25 o C, subculture medium pH 5.8, subculture once every 28 day...

Embodiment 3

[0029] A method for culturing callus cells of Glycyrrhiza glabra that can increase the content of licorice flavonoids comprises the following steps:

[0030] 1) Induction and subculture of Glycyrrhiza glabra callus: Select plump Glycyrrhiza glabra seeds and culture them on MS basal medium containing 30g / L sucrose and 7g / L agar to obtain sterile seedlings; take sterile seedlings Hypocotyls were explants, inserted into MS basal medium containing 1.4mg / L naphthaleneacetic acid, 0.1mg / L 6-benzylaminopurine, 26g / L sucrose, and 5g / L agar, cultured in the dark for 10 days, and induced callus, after which the light intensity was 1500 lx The culture was continued for 14 days under the condition of 12 hours of light every day; then the callus was transferred to the culture medium containing 1.4mg / L naphthaleneacetic acid, 0.1mg / L 6-benzylaminopurine, 0.6mmol / L methionine, 26g / L In the MS basal medium of sucrose and 5g / L agar, the subculture temperature was 25 o C, subculture medium pH...

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Abstract

The invention relates to a glycyrrhiza glabra callus cell culture method being capable of improving content of licoflavone. The method is characterized by including the steps of: (1) choosing full glycyrrhiza glabra seeds, and culturing the seeds on a MS basic culture medium containing 30 g / L of saccharose and 7 g / L of agar to obtain sterile seedlings, and inoculating the sterile seedlings on a callus induction culture medium for cultivation, and moving the sterile seedlings on a subculture medium for subculture; (2) selecting slight-yellow callus after the subculture in the step (1), and transferring the callus on a liquid culture medium to perform suspended shaking cultivation; (3) performing inductive cultivation during the suspended shaking cultivation process of the callus, and increasing culturing temperature from 25 DEG C to 35-45 DEG C from the sixth to twelfth day of the subculture period, and continuously culturing the callus for 1-3 days, reducing the culturing temperature to 25 DEG C, wherein coronatine is added during the cultivation at 35-45 DEG C and p-hydroxyphenylpyruvic acid is added when the temperature is reduced to 25 DEG C, and after the suspended shaking cultivation is finished, the glycyrrhiza glabra callus cell having high content of licoflavone can be obtained. The method achieves high content of the licoflavone.

Description

technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a method for culturing callus cells of licorice glabra that can increase the content of licorice flavonoids. [0002] technical background [0003] Glycyrrhiza glabra is mainly distributed in northern Xinjiang, Qinghai, Gansu and other provinces and regions, and its main chemical components are flavonoids and triterpene saponins. Among them, licorice flavonoids have a variety of biological activities. Oxidation, anti-cancer, anti-cancer and other aspects also have significant biological activities, and have broad application prospects in medicine, food health care, and cosmetics industries. [0004] With the deepening of the development and utilization of Glycyrrhizae glabra, its demand has soared. Since 90% of the world's licorice raw materials are provided by my country's exports, wild licorice has been destructively excavated in most areas of our country, and some areas...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12P17/06
CPCC12N5/04C12P17/06
Inventor 黄洁芳张金龙孙晓昂镇晓华吴依娜邱玉想蒋丽刚
Owner PROYA COSMETICS
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