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A method for isolating cotton guard cell protoplasts

A technology of protoplasts and guard cells, applied to plant cells, etc., to achieve the effects of improving experimental efficiency, shortening experimental time, and high physiological vitality

Active Publication Date: 2017-01-11
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, there is currently no description of the isolation and purification of cotton guard cell protoplasts, and a recent report on the isolation of cotton mesophyll cell protoplasts (Li Nina, Acta Crops 2014, 40(2): 231−239)

Method used

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  • A method for isolating cotton guard cell protoplasts
  • A method for isolating cotton guard cell protoplasts
  • A method for isolating cotton guard cell protoplasts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] A method for isolating cotton guard cell protoplasts, comprising the following steps:

[0024] (1) Take cotton seeds and induce germination;

[0025] In order to break the dormancy of cotton seeds, soak the mature and plump cotton seeds in a water bath at 80°C for 2 minutes;

[0026] Cotton seeds were induced to germinate under the conditions of 16 h light / 8 h dark, light intensity 22 KLux, temperature 20-29°C, and relative humidity 65%;

[0027] (2) Take the cotton true leaves germinated 3-4 weeks after step (1), rinse them with sterile water, tear off the epidermal strips, transfer the collected epidermal strips into the enzymatic hydrolysis solution, and shake in a water bath at 23-28°C 1.5~2 hours free guard cell protoplasts, 20 times microscopic results before and after enzymolysis are as follows figure 1 shown;

[0028] (3) Add 5 to 10 times the volume of the basic medium to the mixed solution after the enzymatic hydrolysis in step (2), filter with a 400-mesh f...

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Abstract

The invention particularly relates to a method for separating a cotton guard cell protoplast. The method comprises the following steps: inducing germination of cotton seeds, carrying out enzymolysis and separation on the guard cell protoplast, centrifuging and the like. According to the method for separating the cotton guard cell protoplast provided by the invention, guard cells of cotton leaves are processed by adopting a one-step enzymolysis approach. Compared with a two-step enzymolysis approach, the experimental period is greatly shortened, the experimental operation process is simplified, the experiment efficiency is improved, high physical activity of the protoplast is kept, the problem of separation of the cotton guard cell protoplast is solved, and a good experimental foundation is provided for research of guard cell permeability, ion transportation and the mechanism of stomatal movement by successful separation of the cotton guard cell protoplast.

Description

technical field [0001] The invention specifically relates to a method for isolating cotton guard cell protoplasts. Background technique [0002] Cotton, a plant of the Malvaceae family of the genus Gossypium, with shrubby plants. Cotton is one of the most important crops in the world, and my country is also one of the countries with the highest cotton production. The main and by-products of cotton have high utilization value. It is not only the most important fiber crop, but also an important oil crop, a high-protein food crop, and a textile, fine chemical raw material and important strategic material. [0003] As an important economic crop and fiber crop, cotton has broad prospects for applying modern biotechnology to cotton breeding and production. In recent years, cotton biotechnology has developed rapidly, and great progress has been made in the cultivation of hybrid plants, the screening of resistant mutants, and the cultivation of transgenic plants resistant to pests...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 张骁赵翔孔培涛王雪娇高歌
Owner HENAN UNIVERSITY
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