Method for artificially creating plants male sterility

A male sterility and plant technology, applied in botany equipment and methods, plant products, plant gene improvement, etc., can solve problems such as interfering with anthers, antisense fragments are difficult to reach the action site, and abortion traits cannot be stably inherited

Inactive Publication Date: 2008-09-10
LIAOCHENG UNIV
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  • Abstract
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Problems solved by technology

But this method is still not completely free from the influence of the Mariani method
[0005] The third method is to use antisense RNA technology or other related technologies to interfere with the normal gene expression during anther (pollen

Method used

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  • Method for artificially creating plants male sterility
  • Method for artificially creating plants male sterility
  • Method for artificially creating plants male sterility

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Experimental program
Comparison scheme
Effect test

Embodiment

[0030] 1. Construction of expression vector

[0031] 1.1 Cloning of rice anther-specific promoter Osg6B

[0032] According to the Osg6B sequence reported in the literature (Tsuchiya and Toriyama et al., 1994) (the sequence shown in SEQ ID NO: 1, NCBI accession number: D21160), the primers were designed using DNASTAR software and synthesized and amplified by Beijing Saibaisheng Biotechnology Company Primers: 6B F-EcoRI 5'ggg gaa ttc ttt ttt tta cac ag3' (SEQ ID NO:2), 6B R-NcoI 5'cca tgg tgg att aga gct tg-3' (SEQ ID NO:3). The genomic DNA of the japonica rice variety Nipponbare was used as a PCR amplification template for PCR amplification. The amplification program is: 94°C 5min; 94°C 45sec, 56°C 45sec, 72°C 2min, 35r; 72°C 5min. (Taq enzyme used in PCR was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., and dNTPs were purchased from Promega Company). After the amplification was completed, the agarose gel (agarose was purchased from Beijing Baierdi Biote...

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Abstract

The invention discloses a method of male sterility in plant by artificial hybridization, which is characterized in that: the method is a method which inhibits the expression of an endogenous HSP70 gene of plant anther through the antisense RNA technology, and interferes the normal growth of the transgenic plant anther so as to acquire a male sterility transgenic plant. In particular, the method is a method which transforms an antisense RNA expression vector of the HSP70 gene driven by an Osg6B promoter expressed by anther specificity into a graminaceous plant, inhibits the expression of the endogenous HSP70 gene of the anther, and interferes the normal growth of the transgenic plant anther so as to acquire the male sterility transgenic plant. The transgenic plant acquired by the method can implement male complete sterility.

Description

technical field [0001] The invention relates to a method for artificially creating male sterility of plants. Specifically, it relates to an antisense RNA expression vector of HSP70 gene driven by anther-specifically expressed Osg6B promoter, which is transferred into Gramineae grain crops to suppress the expression of endogenous HSP70 gene, thereby interfering with the anther growth of transgenic plants. A method for achieving male sterility through normal development, which can lay the foundation for the utilization of heterosis in plants. Background technique [0002] Plant male sterility is the basis of plant heterosis utilization. Usually, the way to obtain plant male sterility is to screen out male sterile mutants in natural resources or through mutagenesis, or through distantly related species. These two methods have disadvantages such as difficulty in obtaining sterile materials and long cycle time. With the development of plant molecular biology and developmental b...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/11A01H1/00A01H5/00
Inventor 刘立科刘春光刘根齐侯宁张文会陈建南
Owner LIAOCHENG UNIV
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