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Method of increasing suspendible cultured embryonic callus induction ratio of momordica grosvenori

A technology of embryogenic callus and suspension culture, applied in horticultural methods, botanical equipment and methods, plant cells, etc., can solve the problems of low induction rate, poor cell performance, and elongated liquid suspension culture system time, etc., to achieve The market prospect is broad, the method is simple, and the effect of shortening the induction time

Active Publication Date: 2017-12-08
GUILIN NATURAL INGREDIENTS CORP
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  • Claims
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AI Technical Summary

Problems solved by technology

The most important thing about the above two methods is to establish a liquid suspension culture system, which requires obtaining a large number of embryogenic callus in good condition, but there are few related studies on the embryogenic callus of Luo Han Guo cells
[0004] Existing Momordica grosvenori embryogenic callus induction is mainly through seedlings after seed germination, part of the stem, leaves and roots of the plant are used as explants to induce embryogenic callus, but the stage of seed germination is It takes at least one month, which greatly lengthens the time to establish a liquid suspension culture system
Moreover, the induction rate of the existing embryogenic callus is also relatively low, and the performance of the induced cells in suspension culture is poor.

Method used

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  • Method of increasing suspendible cultured embryonic callus induction ratio of momordica grosvenori
  • Method of increasing suspendible cultured embryonic callus induction ratio of momordica grosvenori
  • Method of increasing suspendible cultured embryonic callus induction ratio of momordica grosvenori

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Effect test

Embodiment 1

[0029] The method for improving the induction rate of embryogenic callus of Luo Han Guo in suspension culture of the present embodiment comprises the following steps:

[0030] After sterilizing 140 Luo Han Guo seeds, shell them, take the seed embryos, and cut them into sections to obtain 410 seed embryo explants, put them into the induction medium with a pH value of 5.0, and store them at a temperature of 20°C without light. After 21 days of induction culture, 282 embryogenic callus cell masses of Luo Han Guo were obtained, wherein the induction medium was composed of MS basal saline solution medium, sucrose 25g / L, agar 5g / L, inositol 80mg / L, 6 - Composition of BA 2mg / L, NAA 1mg / L and betaine 6g / L. Wherein, the MS basal saline solution culture medium is made of KNO 3 1900mg / L, NH 4 NO 3 1650mg / L, MgSO 4 ·7H 2 O 370mg / L, KH 2 PO 4 170mg / L, CaCl 2 2H 2 O 440mg / L, MnSO 4 4H 2 O 22.3mg / L, ZnSO 4 ·7H 2 O 8.6mg / L, H 3 BO 3 6.2mg / L, KI 0.83mg / L, Na 2 MoO 4 2H 2 O 0....

Embodiment 2

[0037] The method for improving the induction rate of embryogenic callus of Luo Han Guo in suspension culture of the present embodiment comprises the following steps:

[0038] After sterilizing 140 Luo Han Guo seeds, shell them, take the seed embryos, cut them into sections, and obtain 410 seed embryo explants, put them into the induction medium with a pH value of 6.0, and keep the temperature at 25°C without light After 18 days of induction culture, 385 embryogenic callus cell masses of Luo Han Guo were obtained, wherein the induction medium was composed of MS basal saline solution medium, sucrose 30g / L, agar 4.6g / L, inositol 100mg / L, 6-BA 1mg / L, NAA 1mg / L and betaine 5g / L. Wherein, the MS basal saline solution culture medium is made of KNO 3 1900mg / L, NH 4 NO 3 1650mg / L, MgSO 4 ·7H 2 O 370mg / L, KH 2 PO 4 170mg / L, CaCl 2 2H 2 O 440mg / L, MnSO 4 4H 2 O 22.3mg / L, ZnSO 4 ·7H 2 O 8.6mg / L, H 3 BO 3 6.2mg / L, KI 0.83mg / L, Na 2 MoO 4 2H 2 O 0.25mg / L, CuSO 4 ·5H 2 O...

Embodiment 3

[0041] The method for improving the induction rate of embryogenic callus of Luo Han Guo in suspension culture of the present embodiment comprises the following steps:

[0042] After sterilizing 140 Luo Han Guo seeds, shell them, take the seed embryos, cut them into sections, and obtain 410 seed embryo explants, put them into the induction medium with a pH value of 7.0, and store them at a temperature of 30°C without light. After 15 days of induction culture, 304 embryogenic callus cell masses of Luo Han Guo were obtained, wherein the induction medium was composed of MS basal saline solution medium, sucrose 35g / L, agar 4g / L, inositol 160mg / L, 6 -BA 0.5mg / L, NAA 3mg / L and betaine 4g / L. Wherein, the MS basal saline solution culture medium is made of KNO 3 1900mg / L, NH 4 NO 3 1650mg / L, MgSO 4 ·7H 2 O 370mg / L, KH 2 PO 4 170mg / L, CaCl 2 2H 2 O 440mg / L, MnSO 4 4H 2 O 22.3mg / L, ZnSO 4 ·7H 2 O 8.6mg / L, H 3 BO 3 6.2mg / L, KI 0.83mg / L, Na 2 MoO 4 2H 2 O 0.25mg / L, CuSO ...

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Abstract

The invention discloses a method of increasing suspendible cultured embryonic callus induction ratio of momordica grosvenori and belongs to the technical field of industrial biology. The method comprises the following steps: after disinfecting momordica grosvenori seeds, shelling the momordica grosvenori seeds and taking embryos; inoculating the momordica grosvenori seeds into an induction culture medium, the pH value of which is 5-7; and performing inductive culture for 15-21 days at 20-30 DEG C without illumination to obtain the embryonic callus of momordica grosvenori, wherein the inductive culture is composed of an MS basic salt culture medium, 25-35g / L of saccharose, 4-5g / L of agar, 80-120mg / L of inositol, 0.5-2mg / L of 6-BA, 0.5-1.5mg / L of NAA and 4-6g / L of glycine betaine. The induction rate of the suspendible cultured embryonic callus is 64-93.6%, the induction time can be shortened, and calluses which are good in state can be provided for large-scaled culture of momordica grosvenori cells.

Description

technical field [0001] The invention relates to a method for increasing the induction rate of embryogenic callus capable of suspension culture of Luo Han Guo, belonging to the field of industrial biotechnology. Background technique [0002] Luo Han Guo (Siraitia grosvenorii) is derived from the genus Momordica of Cucurbitaceae, and is a precious medicinal and edible plant unique to Guilin, Guangxi. Mogroside is the main active ingredient in Luo Han Guo, which has the functions of clearing away heat and nourishing the lungs, relieving the throat and opening the tone, moistening the intestines and defecating. Recent studies have shown that Luo Han Guo also has antioxidant, anti-diabetic and anti-cancer effects. Luo Han Guo is deeply loved by people for its novel taste, pure taste and unique health functions. It is the best sweetener and health product for patients with obesity, high blood pressure and diabetes, and has a wide range of application values. [0003] The existin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04
CPCA01H4/001A01H4/005C12N5/04
Inventor 宋云飞郭美锦李佳瑞王泽建庄英萍李宇钰
Owner GUILIN NATURAL INGREDIENTS CORP
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