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Method for remarkably improving resistance of cotton to greensickness through verticillium dahlia interference VdRGS1 gene expression

A Verticillium dahliae, cotton technology, applied in the direction of genetic engineering, plant genetic improvement, chemical instruments and methods, can solve functional analysis and phenotypic identification has not been reported, has not been applied to the breeding practice of improving cotton resistance, no Issues such as being verified

Inactive Publication Date: 2018-01-16
NANJING AGRICULTURAL UNIVERSITY
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  • Application Information

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Problems solved by technology

[0007] So far, the functional analysis and phenotypic identification of RGS in Verticillium dahliae have not been reported, and have not been applied to the breeding practice of improving cotton resistance to Verticillium dahliae
At the same time, VIGS-based HIGS technology has not yet been verified in the interaction between cotton and Verticillium wilt

Method used

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  • Method for remarkably improving resistance of cotton to greensickness through verticillium dahlia interference VdRGS1 gene expression
  • Method for remarkably improving resistance of cotton to greensickness through verticillium dahlia interference VdRGS1 gene expression
  • Method for remarkably improving resistance of cotton to greensickness through verticillium dahlia interference VdRGS1 gene expression

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Embodiment 1

[0050] (1) Discovery and functional analysis of Verticillium dahliae VdRGS1

[0051] Using the reported genome information of Verticillium dahliae (http: / / genome.jgi.doe.gov / Verda1 / Verda1.home.html), the RGS system was analyzed, and 8 RGS genes were found after verification. Based on the above sequence information, primers were designed for further homologous cloning in Verticillium dahliae Vd8. Cotton seedling roots were treated with Vd8 bacterial solution for 0, 2, 4, 6, 12 and 24 hours, and the expression characteristics of 8 RGS genes were analyzed at different times when Verticillium dahliae was exposed to cotton seedling root tissues. It was found that only the VdRGS1 gene had the highest expression level and the most obvious induced expression. After 12 and 24 hours of contact with cotton roots, the expression level increased by 20-30 times, suggesting that this gene plays an important role in the pathogenicity of Verticillium dahliae ( figure 1 ). The primers used in...

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Abstract

The invention discloses a method for remarkably improving resistance of cotton to greensickness through verticillium dahlia interference VdRGS1 gene expression and belongs to the field of biological technique application. The invention provides a nucleotide sequence of a VdRGS1 gene and an ORF sequence and an amino acid sequence of cDNA. Function and phenotypic analysis shows that VdRGS1 is capable of regulating and controlling pathogenicity of verticillium dahlia, formation of microsclerotia, growth of spores and hyphae, and the like, and plays a significant role when verticillium dahlia invades and infect a host. By using a transgenic technique, a greensickness bacterium which can express green fluorescent protein (GFP) is created, by using a virus-induced gene silencing (VIGS) technique, a GFP gene of the transgenosis verticillium dahlia can be efficiently silenced in cotton, the result shows that host induced gene silencing (HIGS) is achieved on the basis of the VIGS technique, andinvading verticillium dahlia genes can be successfully silenced in cotton by using the technique. By using an HIGS technique based on VIGS, the verticillium dahlia VdRGS1 is silenced in cotton, and the resistance of cotton to verticillium dahlia is remarkably improved.

Description

technical field [0001] The invention relates to a VdRGS1 gene of Verticillium dahliae and a method for significantly improving the resistance of cotton to Verticillium dahliae by interfering with the expression of the VdRGS1 gene. Verticillium dahliae Vd8 and V991 are two highly pathogenic strains that infect cotton, causing the cotton Verticillium wilt phenotype. RGS is a negative regulator of the G protein α subunit in the G protein signaling pathway. The VdRGS1 gene in Verticillium dahliae Vd8 was knocked out and complemented by the method mediated by Agrobacterium, and the knockout and complementation mutants of VdRGS1 were obtained. Functional and phenotypic analysis showed that VdRGS1 regulates the pathogenicity, microsclerotia formation, spore and hyphae growth of Verticillium dahliae, and plays an important role in the infection of the host by Verticillium dahliae. By creating a transgenic V991 Verticillium dahliae expressing green fluorescent protein (GFP), it was ve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/37C12N15/82A01H6/60
Inventor 郭旺珍徐君
Owner NANJING AGRICULTURAL UNIVERSITY
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