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Method for separating cotton guard cell protoplast

A protoplast and guard cell technology, applied in plant cells and other directions, can solve the problem of no description of separation and purification of cotton guard cells, and achieve the effect of shortening the experimental time, improving the experimental efficiency, and simplifying the experimental operation process.

Active Publication Date: 2014-11-19
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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  • Method for separating cotton guard cell protoplast
  • Method for separating cotton guard cell protoplast
  • Method for separating cotton guard cell protoplast

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] The method for separating cotton guard cell protoplasts includes the following steps:

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

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

[0026] Cotton seeds were induced to germinate under the conditions of 16 hours of light / 8 hours of darkness, light intensity of 22 KLux, temperature of 20~29℃, and relative humidity of 65%;

[0027] (2) Take the true cotton leaves 3~4 weeks after germination in step (1), rinse with sterile water, tear off the epidermal strips, transfer the collected epidermal strips into the enzymatic hydrolysis solution, and shake in a 23~28℃ water bath Free guard cell protoplasts within 1.5~2 hours, and the 20-fold microscopic results before and after enzymolysis are as follows figure 1 Shown

[0028] (3) Add 5-10 times the volume of the basic medium to the mixed solution after the enzymolysis in step (2), filter w...

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PUM

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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 separating cotton guard cell protoplasts. Background technique [0002] Cotton, cotton plant of Malvaceae, shrub-like 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, textile, fine chemical raw materials, and important strategic materials. [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 that are resistant to pests and diseases. [0004] Guard ce...

Claims

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Application Information

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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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