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A kind of culture method of golden gentian grass in vitro cells

The technology of a golden gentian grass and a cultivation method, which is applied in the field of plant tissue culture, can solve the problems that resources cannot meet the needs of the pharmaceutical market and scientific research, and achieve the effect of short cycle.

Active Publication Date: 2022-08-02
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the existing golden gentian herbs are collected in the wild and artificially cultivated. Due to the influence of natural factors and cultivation cycles, their resources are far from meeting the needs of the medical market and scientific research.

Method used

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  • A kind of culture method of golden gentian grass in vitro cells
  • A kind of culture method of golden gentian grass in vitro cells
  • A kind of culture method of golden gentian grass in vitro cells

Examples

Experimental program
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Effect test

Embodiment 1

[0040] A method for in vitro rapid propagation and suspension cells of Gentianae chinensis comprises the following steps:

[0041] Step 1. Selection and sterilization of explants: collect new leaves with robust growth and no pests and diseases as spare explants. In a clean bench, the explants were rinsed under sterile water, and the residual water was blotted with sterile filter paper. The treated leaves were soaked in 70% alcohol for 10s, rinsed with plenty of sterile water for 3-5 times immediately after taking them out, and then put in 0.1% HgCl 2 Soak for 8 minutes, rinse with plenty of sterile water 3-5 times immediately after the end, then place the explants on sterile filter paper to absorb water, and use a sterile scalpel to cut off the leaf veins;

[0042] Step 2. Separation of single cells from leaves of Gentiana chinensis:

[0043] Use mechanical method to separate single cells, transfer the treated leaves into 20ml tripe homogenizer (sterile) containing STN buffe...

Embodiment 2

[0049] A method for in vitro rapid propagation and suspension cells of Gentianae chinensis comprises the following steps:

[0050] Step 1. Selection and sterilization of explants: collect new leaves with robust growth and no pests and diseases as spare explants. In a clean bench, the explants were rinsed under sterile water, and the residual water was blotted with sterile filter paper. The treated leaves were soaked in 70% alcohol for 30s, immediately rinsed with plenty of sterile water for 3-5 times, and then put in 0.1% HgCl 2 Soak for 6 minutes, rinse immediately with plenty of sterile water 3-5 times, then place the explants on sterile filter paper to absorb water, and use a sterile scalpel to cut off the leaf veins;

[0051] Step 2. Separation of single cells from leaves of Gentiana chinensis:

[0052] Use mechanical method to separate single cells, transfer the treated leaves into 20ml tripe homogenizer (sterile) containing STN buffer, quickly grind and homogenize for ...

Embodiment 3

[0058] A method for in vitro rapid propagation and suspension cells of Gentianae chinensis comprises the following steps:

[0059] Step 1. Selection and sterilization of explants: collect new leaves with robust growth and no pests and diseases as spare explants. In a clean bench, the explants were rinsed under sterile water, and the residual water was blotted with sterile filter paper. The treated leaves were soaked in 70% alcohol for 20s, immediately rinsed with plenty of sterile water for 3-5 times, and then put in 0.1% HgCl 2 Soak for 10 minutes, rinse immediately with plenty of sterile water 3-5 times, then place the explants on sterile filter paper to absorb water, and use a sterile scalpel to cut off the leaf veins;

[0060] Step 2. Separation of single cells from leaves of Gentiana chinensis:

[0061] Single cells were separated by mechanical method, and the treated leaves were transferred into 20ml tripe homogenizer (sterile) containing STN buffer, quickly ground and...

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Abstract

The invention discloses a method for culturing in vitro cells of Gentiana chinensis, comprising the following steps: selection and sterilization of explants; separation of single cells from leaves of Gentiana chinensis; establishment of a suspension cell line of Gentiana chinensis; and stable subculture. The cycle of the invention is shorter than that of tissue culture, and at the same time, it does not need to cultivate golden gentian, and is not limited by geographical factors.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, in particular to a method for culturing in vitro cells of Gentianae chinensis. Background technique [0002] Golden gentian (Conyzablinii H.Lév.) is a biennial herb of the Compositae Liquoraceae, also known as bear bile, etc. It grows in the sparse forest area at an altitude of about 2000 meters, mainly distributed in the Panxi area of ​​Sichuan, central and southern Yunnan and parts of western Guizhou. As a medicinal plant, it is cold in nature and bitter in taste, and has the functions of clearing heat, anti-inflammatory, detoxifying, removing stasis, and reducing swelling. Significant curative effect. Most of the existing golden gentian herbal materials are collected in the wild and cultivated artificially. Due to the influence of natural factors and cultivation cycles, their resources are far from meeting the needs of the pharmaceutical market and scientific research. [0003]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/04
Inventor 陈惠郑天润孙文君詹俊义刘燕汪茂佳刘默洋马招堂王涛唐自钟布同良吴琦李成磊
Owner SICHUAN AGRI UNIV