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Optimization method of wheat pollen cell electric-shock-method transformation system and application of optimization method

A technology for wheat pollen and optimization methods, applied in the direction of biochemical equipment and methods, applications, botany equipment and methods, etc., can solve the problems of low seed setting rate, high requirements, and long time consumption, and achieve low cost, simple operation, and future generations The effect of stable homozygous fast

Inactive Publication Date: 2016-03-30
NORTHWEST A & F UNIV
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Problems solved by technology

[0012] The purpose of the present invention is to provide an optimization method and application of an electric shock transformation system for wheat pollen cells, aiming to solve the problems of cumbersome, time-consuming, high requirements and low seed-setting rate when transforming by means of tissue culture

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  • Optimization method of wheat pollen cell electric-shock-method transformation system and application of optimization method
  • Optimization method of wheat pollen cell electric-shock-method transformation system and application of optimization method
  • Optimization method of wheat pollen cell electric-shock-method transformation system and application of optimization method

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[0084] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0085] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0086] Such as figure 1 As shown, the optimization method and application of the wheat pollen cell electric shock transformation system of the embodiment of the present invention include the following steps:

[0087] S101: Extracting plasmids containing DREB4A and W17 genes from the bacterial liquid;

[0088] S102: Optimization of the optimum electric shock solution for wheat pollen;

[0089] S103: FDA staining and observation and statistics after electric shock;

[0090] S104: Opt...

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Abstract

The invention discloses an optimization method of a wheat pollen cell electric-shock-method transformation system and application of the optimization method. The optimization method takes a wheat ripe pollen cell as a transformation acceptor; electric shock conditions of electric shock liquid and germination liquid are screened and optimized so that the vitality and in-vitro germination rate of the wheat ripe pollen cell are improved; by using an electric shock transformation method, transcription factor genes related to dry, high-salt and low-temperature induced expression are transformed to obtain transgenic plants. Compared with current main transgenic methods including an agrobacterium transformation method and a gene gun transformation method and the like, a pollen electric shock transformation method is simple to operate and has relatively low cost, a tissue culture process is omitted and the problems that the reproducibility is not strong and a chimera is easy to generate are solved; the acceptor has no gene type limitation and the transformed plants have the advantages of stable later generations, rapidness in homozygosis and the like; a resistance screening process in the tissue culture process does not need to be carried out and the safe transgenic plants without screening markers can be obtained.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering breeding, and in particular relates to an optimization method for the transformation system of wheat pollen cells by electroshock method, and the application of this method in obtaining retrograde-resistant genetically modified wheat. Background technique [0002] Wheat is one of the main food crops, and its total output is only lower than that of corn. So far, the cultivation of new wheat varieties in my country is mainly through the method of conventional hybrid breeding. However, this method has a long cycle, narrow selection range, poor predictability, and low efficiency of trait improvement, thus delaying the development of wheat breeding. [0003] Compared with conventional breeding, molecular breeding shortens the breeding period, provides the possibility of genetic exchange between different species, makes up for the shortcomings of conventional breeding, and has been widely u...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00
CPCC12N15/8273C12N15/8206
Inventor 张小红闵东红邵景侠黎飞飞赵雪晶邓艳芳胡晓薇田原付金梅乔梦王晓东普正菲
Owner NORTHWEST A & F UNIV
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