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A method for promoting the growth of Luo Han Guo suspended cells and increasing the content of glucoside v by salt stress

A suspension cell and Luo Han Guo technology, applied in plant cells, fermentation and other directions, to achieve the effect of short culture cycle time, increased biomass and high growth rate

Active Publication Date: 2022-06-21
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At home and abroad, a lot of research has been done on the physiological, biochemical, and molecular biological response mechanisms of trees to salt, but there is no report on the effect of salt stress and potassium-sodium ratio on the suspension cells of Luo Han Guo

Method used

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  • A method for promoting the growth of Luo Han Guo suspended cells and increasing the content of glucoside v by salt stress
  • A method for promoting the growth of Luo Han Guo suspended cells and increasing the content of glucoside v by salt stress
  • A method for promoting the growth of Luo Han Guo suspended cells and increasing the content of glucoside v by salt stress

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Step 1: Take the mature seed embryo of Luo Han Guo, put it in B5 containing sucrose 25g / L, 6-benzylaminoadenine 0.05mg / L, naphthalene acetic acid 2mg / L, inositol 50mg / L, agar 4g / L, pH 5.9 Subculture on solid medium, select fresh embryogenic callus that is continuously subcultured for 3 times and has vigorous growth, loose texture and stable and uniform state; step 2: the fresh embryogenic callus obtained in step 1 is inoculated with an amount of 25g / L was transferred to B5 liquid medium containing sucrose 20g / L, 6-benzylaminoadenine 0.05mg / L, naphthalene acetic acid 3mg / L, inositol 50mg / L, pH 5.9, and the speed was 80r / L. min, the culture temperature was 24°C, and the cells were cultured in a dark shaker to obtain the first-generation Luo Han Guo suspension cell system.

[0034] Step 3: Cultivate mature Luo Han Guo suspension cells with the primary-generation Luo Han Guo suspension cell system obtained in step 2; the potassium-sodium ratio in the original medium is 25:...

Embodiment 2

[0038] Step 1: Take the seed embryo of Luo Han Guo, put it in a solid B5 containing sucrose 35g / L, 6-benzylaminoadenine 0.15mg / L, naphthalene acetic acid 6mg / L, inositol 150mg / L, agar 6g / L, pH 6.0 Subculture on the medium, and select the embryogenic callus that has been subcultured 5 times in succession;

[0039]Step 2: The embryogenic callus obtained in step 1 was transferred to the inoculum containing sucrose 40g / L, 6-benzylaminoadenine 0.15mg / L, naphthalene acetic acid 5mg / L, inositol 150mg / L at an inoculum amount of 75g / L , pH 6.0 B5 liquid medium, and cultured in a shaker with a rotating speed of 150 r / min, a culture temperature of 26 °C, and a dark shaker to obtain the first-generation Luo Han Guo suspension cell system.

[0040] Step 3: Cultivate mature Luo Han Guo suspension cells with the primary-generation Luo Han Guo suspension cell system obtained in step 2; the potassium-sodium ratio in the original medium is 25:1, the potassium ion concentration is controlled to ...

Embodiment 3

[0044] Step 1: Take the mature seed embryos of Luo Han Guo, in a mixture containing sucrose 30g / L, 6-benzylaminoadenine 0.1mg / L, naphthalene acetic acid 4.0mg / L, inositol 100mg / L, agar 5.0g / L, pH range of Subculture on the B5 solid medium of 5.9, select fresh embryogenic callus with 4 consecutive subcultures and vigorous growth, loose texture, and stable and uniform state;

[0045] Step 2: The fresh embryogenic callus obtained in step 1 was transferred to the inoculum containing sucrose 30g / L, 6-benzylaminoadenine 0.1mg / L, naphthalene acetic acid 4.0mg / L, inositol 100mg at an inoculum amount of 50g / L / L B5 liquid medium with pH range of 5.9, and cultured in a shaker with a rotating speed of 110r / min, a culture temperature of 25°C, and a dark shaker to obtain the first-generation Luo Han Guo suspension cell system;

[0046] Step 3: Cultivate mature Luo Han Guo suspension cells with the primary-generation Luo Han Guo suspension cell system obtained in step 2; the potassium-sodiu...

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Abstract

The invention relates to a method for promoting the growth of Luo Han Guo suspension cells and increasing the content of sweet glycoside V through salt stress, which is characterized in that the concentration of potassium ions is controlled to be 100-2000 mg / kg during the process of cultivating mature Luo Han Guo suspension cells with the first-generation Luo Han Guo suspension cell system , The concentration ratio of potassium and sodium ions is 5-20:1. The method of the present invention improves the growth amount, specific growth rate, mogroside V yield and synthesis rate of Luo Han Guo, is beneficial to the growth of Luo Han Guo suspension cells and the market-oriented large-scale production of Mogroside V; at the same time, the production and cultivation period of cells is effectively shortened, Artificially adjusting the production process of Momordica grosvenori suspension cell biomass and mogroside V is beneficial to the quality control and management of products.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for promoting the growth of Luo Han Guo suspended cells and increasing the content of glucoside V through salt stress. Background technique [0002] Mogroside V, also known as mogroside, is the main sweet component of Luo Han Guo, the genus of cucurbitaceae, and its sweetness is 350 times that of sucrose. Seven monomer components have been identified, among which mogroside V and S. Gateside I has the strongest sweetness. Mogroside Ⅴ has medicinal effects such as clearing away heat and moistening the lungs, soothing the throat and opening the sound, and moistening the bowels and laxatives. It is the most suitable sweetener and health care product for special groups such as patients with diabetes, hypertension and diabetes, and has a wide range of application value. At present, the method for obtaining Mogroside V is mainly made from natural Luo Han Guo, which is extracted a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C12P33/00
CPCC12N5/04C12P33/00
Inventor 郭美锦宋云飞陈雨霞王泽建李杰杨文国庄英萍储炬肖慈英
Owner EAST CHINA UNIV OF SCI & TECH
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