Application of protein FoAtg27 in regulation of pathogenicity of fusarium oxysporum

A technology of banana fusarium wilt and banana wilt, which is applied in the field of plant genetic engineering and can solve problems such as unclear specific functions

Active Publication Date: 2022-07-29
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The specific function of FoAtg27 in Fusarium wilt of banana is still unclear

Method used

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  • Application of protein FoAtg27 in regulation of pathogenicity of fusarium oxysporum
  • Application of protein FoAtg27 in regulation of pathogenicity of fusarium oxysporum
  • Application of protein FoAtg27 in regulation of pathogenicity of fusarium oxysporum

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Experimental program
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Effect test

Embodiment 1

[0038] 1 Experimental material

[0039] 1.1 Test strains and plants

[0040] The test strain was Fusarium oxysporum f.sp. cubense 4 (Foc4), and the test plant was Brazil banana (Cavendish, AAA) with 4-5 leaves.

[0041] 1.2 Host bacteria and plasmid vector

[0042] The host strain is Escherichia coli DH5α strain. The cloning vector is pMD18-T vector, the gene knockout vector is the filamentous fungus expression vector pCT74, and the gene complementing vector is pCTZN (which was transformed from the pCT74 plasmid in our laboratory, that is, the gfp and hph genes on pCT74 were replaced with bo lyomycin (Zeocin) gene).

[0043] 2 Experimental methods

[0044] 2.1 Amplification of upstream and downstream homologous fragments of FoAtg27 gene

[0045] The construction of Fusarium wilt FoAtg27 gene knockout vector is as follows figure 1 shown. The upstream and downstream sequences of the FoAtg27 gene with a length of about 1500 bp (named as homology arm A fragment and homology a...

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Abstract

The invention discloses application of a protein FoAtg27 in regulation and control of pathogenicity of fusarium oxysporum, and belongs to the field of plant genetic engineering. The method comprises the following steps: constructing a gene knockout vector, and introducing the gene knockout vector into a Foc4 protoplast to obtain a knockout mutant delta FoAtg27; the method comprises the following steps: constructing a gene back-filling vector, and introducing the gene back-filling vector into a knockout mutant protoplast to obtain a back-filling mutant delta FoAtg27-com. Compared with Foc4, the sporulation quantity of delta FoAtg27 is remarkably reduced, and the sensitivity to Congo red stress is remarkably reduced; a pathogenicity test shows that the deficiency of FoAtg27 enables the pathogenicity of Foc4 to be obviously reduced; after the gene is supplemented, the sporulation quantity and pathogenicity of the gene are recovered. The invention proves that the FoAtg27 gene is necessary for the generation and pathogenicity of Foc4 conidia. The research is helpful to deeply clarify the pathogenic molecular mechanism of Foc4, and a target gene is provided for the development of effective bactericides.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and particularly relates to the application of an autophagy-related protein (Autophagy-related protein 27) FoAtg27 of Fusarium oxysporum in regulating the pathogenicity of Fusarium oxysporum. Background technique [0002] Fusarium wilt of banana (FWB), also known as Panama disease (Panamadisease) or banana yellow leaf disease, is a disease caused by Fusarium oxysporumf.sp.cubense (Foc). This devastating soil-borne disease seriously threatens the development of the global banana industry. Foc can infect different banana varieties. According to this, Foc can be divided into race 1 (Foc1), race 2 (Foc2) and race 4 (Foc4). Among them, Foc4 has the strongest infectivity and is more harmful The largest, can infect almost all banana varieties. This patent contributes to a comprehensive understanding of the composition and function of Foc4 effector proteins, and provides a theoretical basis for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/80C12N15/31A01N47/44A01P3/00C12R1/77
CPCC12N15/80C07K14/37A01N47/44
Inventor 聂燕芳赵雅丽李云锋鄢甜甜李洁玲李华平
Owner SOUTH CHINA AGRI UNIV
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