Method for effectively inducing cotton somatic embryo and regenerating plants and culture medium thereof

A technology of liquid culture medium and culture medium, applied in plant regeneration, botany equipment and methods, gardening methods, etc., can solve the problems of long cycle, poor repeatability, high frequency of deformed embryos, etc.

Inactive Publication Date: 2012-04-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cotton is a crop that is difficult to obtain regenerated plants through somatic embryogenesis, which is characterized by strong genotype dependence, long cycle and poor reproducibility in tissue culture.
In addition, during somatic embryogenesis in cotton, the efficiency of somatic embryogenesis is low and the frequency of malformed embryos is high

Method used

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  • Method for effectively inducing cotton somatic embryo and regenerating plants and culture medium thereof
  • Method for effectively inducing cotton somatic embryo and regenerating plants and culture medium thereof
  • Method for effectively inducing cotton somatic embryo and regenerating plants and culture medium thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1. Efficiently Obtaining Somatic Embryos and Regenerated Plants

[0076] (1) Acquisition of sterile cotyledon seedlings

[0077] 1. Delinting of cotton seeds

[0078] Take 50g of cotton variety Kezimian 201 seeds, add about 15ml of concentrated sulfuric acid, stir quickly for 2-3 minutes, then neutralize with lime water, scrub continuously, and rinse with running water for 10-12 minutes to obtain clean delint seeds.

[0079] 2. Germination of cotton seeds

[0080] The depilled seeds were subjected to 0.1% (w / v) HgCl 2 The solution was sterilized for 10 minutes, then the seeds were washed 5-6 times with sterilized water, and finally 150ml of sterilized water was added, and germinated in the dark at 28°C for 24 hours to obtain germinated seeds.

[0081] 3. Germination of cotton seeds

[0082] Peel off the husks of the germinated seeds in an aseptic environment, plant the husk-removed cotton seed embryos on a germination medium, and culture them in the dark at ...

Embodiment 2

[0107] Example 2. Efficiently Obtaining Somatic Embryos and Regenerated Plants

[0108] (1) Acquisition of sterile cotyledon seedlings

[0109] 1. Delinting of cotton seeds

[0110] Take 50g of cotton variety Jiwu 2031 (purchased from Hebei Jifeng Seed Industry Co., Ltd., a variety approved by Hebei Provincial Crop Variety Approval Committee) seeds, add about 15ml of concentrated sulfuric acid, stir quickly for 2-3min, then neutralize in lime water, scrub continuously, and run water Rinse for 10-12 minutes to obtain clean depilled seeds.

[0111] 2. Germination of cotton seeds

[0112] The depilled seeds were subjected to 0.1% (w / v) HgCl 2 The solution was sterilized for 10 minutes, then the seeds were washed 5-6 times with sterilized water, and finally 150ml of sterilized water was added, and germinated in the dark at 28°C for 24 hours to obtain germinated seeds.

[0113] 3. Germination of cotton seeds

[0114] Peel off the husks of the germinated seeds in a sterile envi...

Embodiment 3

[0135] Example 3. Efficiently Obtaining Somatic Embryos and Regenerated Plants

[0136] (1) Acquisition of sterile cotyledon seedlings

[0137] 1. Delinting of cotton seeds

[0138] Take 50g of cotton variety Nongda 58 (purchased from Hebei Jifeng Seed Industry Co., Ltd., a variety approved by the Hebei Provincial Crop Variety Approval Committee) seeds, add about 15ml of concentrated sulfuric acid, stir quickly for 2-3min, then neutralize with lime water, scrub continuously, and rinse with running water 10-12min to obtain clean fluffed seeds.

[0139] 2. Germination of cotton seeds

[0140] The depilled seeds were subjected to 0.1% (w / v) HgCl 2 The solution was sterilized for 10 minutes, then the seeds were washed 5-6 times with sterilized water, and finally 150ml of sterilized water was added, and germinated in the dark at 28°C for 24 hours to obtain germinated seeds.

[0141] 3. Germination of cotton seeds

[0142] Peel off the husks from the germinated seeds in a steri...

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Abstract

The invention discloses a method for effectively inducing cotton somatic embryo and regenerating plants and culture medium thereof. The culture medium of the invention is applied to the sets of culture medium of the regeneration plants of the cotton somatic cells, and the culture medium comprises culture medium A, culture medium B and culture medium C. The culture medium A comprises basic culture medium of improved MS, hormone, glucose and coagulating agent. The culture medium B comprises the basic culture medium of the improved MS, the hormone, the glucose and amino acid additive. The culture medium C comprises the basic culture medium of the improved MS, the hormone, the glucose, the amino acid additive and the coagulating agent. The method of the invention is a method for cultivating the cotton callus tissues as the regeneration plants by using the sets of the culture medium. The invention cannot be limited by the types of the cotton genes and has short cultivation period and high repeatability, so the cotton effectively obtains a large number of cell embryos and has the ability of regenerating the plants so as to provide an important platform of the researches relative to the cell engineering and the genetic engineering of the cotton and the genetic improvement of the cotton.

Description

technical field [0001] The invention relates to a method for efficiently inducing cotton somatic embryogenesis and plant regeneration and a culture medium thereof. Background technique [0002] Cotton is an important economic crop, and its cell and tissue culture research has made great progress, and the technical system of cotton tissue culture has been initially established and improved. Cotton tissue culture and in vitro regeneration, as the basic technology of cotton cell engineering and genetic engineering, play an extremely important role in cotton genetic improvement. However, cotton is a crop that is difficult to obtain regenerated plants through somatic embryogenesis, which is characterized by strong genotype dependence, long cycle and poor reproducibility in tissue culture. In addition, during somatic embryogenesis in cotton, the efficiency of somatic embryogenesis is low and the frequency of malformed embryos is high. Contents of the invention [0003] The pur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 华金平张园刘正杰王彦霞王玉美
Owner CHINA AGRI UNIV
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