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Application of protein FoUpe2 in regulation and control of pathogenicity of fusarium oxysporum

A technology of banana fusarium wilt and pathogenicity, applied in the field of plant genetic engineering, can solve problems such as unclear function, and achieve the effects of increasing pathogenicity, restoring pathogenicity, and increasing sensitivity

Active Publication Date: 2022-07-22
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 FoUpe2 in Fusarium wilt of banana is still unclear

Method used

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  • Application of protein FoUpe2 in regulation and control of pathogenicity of fusarium oxysporum
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  • Application of protein FoUpe2 in regulation and control of pathogenicity of fusarium oxysporum

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Embodiment 1

[0039] 1 Experimental material

[0040] 1.1 Test strains and plants

[0041] The tested strain was Fusarium oxysporum f.sp. cubense 4 (Foc4), and the tested plant was Brazil banana (Cavendish, AAA) with 4-5 leaves.

[0042] 1.2 Host bacteria and plasmid vectors

[0043] The host strain is Escherichia coli DH5α strain. The cloning vector is pMD18-Tvector, 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 are replaced by bolai Zeocin gene).

[0044] 2 Experimental methods

[0045] 2.1 Amplification of upstream and downstream homologous fragments of FoUpe2 gene

[0046] The construction of Fusarium oxysporum FoUpe2 gene knockout vector is as follows figure 1 shown. The upstream and downstream lengths of the FoUpe2 gene were selected to be 1487 bp and 1049 bp (named as homology arm A fragment and homology...

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Abstract

The invention discloses application of a protein FoUpe2 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; the gene is knocked out from Foc4 by using a homologous recombination method, and a knockout mutant delta FoUpe2 is obtained; the method comprises the following steps: constructing a gene back-up vector, and introducing the gene back-up vector into a delta FoUpe2 protoplast; the gene is supplemented into the knockout mutant by using a random insertion method, so that a supplemented mutant delta FoUpe2-com is obtained. Compared with Foc4, the sensitivity of the delta FoUpe2 to SDS stress is obviously improved; a pathogenicity test shows that the deficiency of FoUpe2 enables the pathogenicity of Foc4 to be obviously improved; after the gene is supplemented, the pathogenicity of the gene is recovered. The research is helpful to deeply clarify the pathogenic molecular mechanism of banana fusarium oxysporum, 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, in particular to the application of an unknown protein (Uncharacterized protein) FoUpe2 of Fusarium wilt of banana in regulating the pathogenicity of Fusarium wilt of banana. 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. Secreted proteins are important pathogenic factors of Foc, but the functions of most of the secreted proteins of Foc are currently unclear. [0003] FoUpe2 (Uncharacterized protein) is an unknown protein that does not contain a known domain and is highly conserved in Fusarium. The specific function of FoUpe2 in Fusarium oxysporum is unclear. This patent contributes to a comprehensive understanding of the composition an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C12N15/31C12N1/15C12Q1/6895A01N57/16A01P3/00C12R1/77
CPCC07K14/37C12Q1/6895A01N57/16C12Q2600/13
Inventor 李云锋聂燕芳赵雅丽鄢甜甜张炎至李华平
Owner SOUTH CHINA AGRI UNIV
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