Use of verticillium dahliae pathogenicity associated protein VdpdaAl

A technology of Verticillium dahliae, pathogenicity, used in applications, fungicides, fungi, etc.

Inactive Publication Date: 2015-09-23
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since no effective control agents and disease-resistant varieties have been developed so far, it is called the "cancer" of cotton. Th...

Method used

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  • Use of verticillium dahliae pathogenicity associated protein VdpdaAl
  • Use of verticillium dahliae pathogenicity associated protein VdpdaAl
  • Use of verticillium dahliae pathogenicity associated protein VdpdaAl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Example 1. Preparation of Verticillium dahliae VdpdaA1 knockout mutant

[0108] 1. Preparation of recombinant Agrobacterium pRF-HU2::PDA1

[0109] Using the genomic DNA of the wild-type strain V592 (V592 strain) of the cotton Verticillium wilt strain (V.dahliae) as a template, PCR amplification was carried out with PDA1up-s and PDA1up-x as primers to obtain PCR amplification The product was named as the left homology arm fragment of the VdpdaA1 gene;

[0110]The genomic DNA of the V592 strain was used as a template, and PDA1down-s and PDA1down-x were used as primers for PCR amplification to obtain a PCR amplification product, which was named the right homology arm fragment of the VdpdaA1 gene.

[0111] PacI and Nt.BbvCI restriction endonucleases were used to double-digest the pRF-HU2 vector ( figure 1 ) to obtain two linearized fragments of the pRF-HU2 vector.

[0112] Ligate the fragment of the left homology arm of the VdpdaA1 gene and the fragment of the right homo...

Embodiment 2

[0150] Example 2. Phenotype of complementary mutants

[0151] 1. Construction of complementary vector

[0152] 1. Take the genomic DNA of the V592 strain as a template, and carry out PCR amplification with PDA1-s and PDA1-x as primers to obtain a PCR amplification product, which is identified as the VdpdaA1 gene by agarose gel electrophoresis ( Figure 9 ), including the start codon ATG and the stop codon TAG, with a size of 988 bp. Using water as a template, the above experiments were performed as a negative control. The nucleotide sequence of the PCR product is the nucleotide sequence shown in positions 36 to 1023 of SEQ ID No. 3.

[0153] 2. Use restriction endonuclease SalI and SpeI double enzyme to cut the PCR amplification product of the VdpdaA1 gene obtained in step 1 to obtain the target gene fragment; insert the target gene fragment into the carrier pSULPH-mut-RG#PB (its nucleotide sequence As shown in SEQ ID No. 6) between the SalI and SpeI recognition sites, othe...

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Abstract

The invention discloses use of verticillium dahliae pathogenicity associated protein VdpdaAl. The use comprises the following steps: knocking out VdpdaAl genes from a wild type verticillium dahliae strain V592 to obtain a VdpdaAl gene knocked out mutant, wherein the sporulation quantity and the cotton pathogenicity of the VdpdaAl gene knocked out mutant are both obviously lower than those of the wild type verticillium dahliae strain V592, which shows that the VdpdaAl genes not only participate in the formation of conidia, but also are a key type of genes of the wild type verticillium dahliae strain V592. The verticillium dahliae pathogenicity associated protein VdpdaA and the related gene are assayed for the first time, so as to provide a foundation for further clarification of the nosogenesis of the verticillium dahliae; the VdpdaAl gene knocked out mutant of the wild type verticillium dahliae strain provides a pathogenic bacterium resource for research of interaction between the verticillium dahliae and a host. The use has an important application value for controlling of cotton verticillium wilt and breeding of a novel cotton variety being resistant to the verticillium dahliae.

Description

technical field [0001] The invention relates to the use of the pathogenicity-related protein VdpdaA1 of Verticillium dahliae in the field of biotechnology. Background technique [0002] Cotton Verticillium wilt is a soil-borne disease of the vascular system, caused by Verticillium dahliae, which causes serious harm to cotton production. Since 1935, cotton Verticillium wilt was introduced into my country with the introduction of cotton seeds, and then began to gradually spread across the country. Especially since the 1990s, cotton verticillium wilt has spread very rapidly in my country. Among them, cotton verticillium wilt was the most serious in 1993, with an incidence area of ​​2,666,700 hm2. 2 , followed by continuous outbreaks nationwide in 1995, 1996, 1999, 2000, 2002, and 2003, resulting in serious losses. Cotton Verticillium wilt has posed a great threat to my country's cotton production and sustainable development. [0003] Cotton verticillium wilt is caused by fung...

Claims

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Application Information

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IPC IPC(8): C12N15/80C12N1/15C07K14/37C12N15/31C12N15/82A01H5/00A01N57/16A01P3/00
Inventor 高峰贾培松毛建才
Owner SHIHEZI UNIVERSITY
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