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Application of gene FoPao in regulation and control of pathogenicity of banana fusarium wilt

A technology of banana fusarium wilt and banana wilt, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2020-08-14
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No related function of polyamine oxidase has been reported in Fusarium wilt

Method used

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  • Application of gene FoPao in regulation and control of pathogenicity of banana fusarium wilt
  • Application of gene FoPao in regulation and control of pathogenicity of banana fusarium wilt
  • Application of gene FoPao in regulation and control of pathogenicity of banana fusarium wilt

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Embodiment 1

[0038] 1. Experimental materials

[0039] 1.1 Tested strains and plants

[0040] Physiological race No. 4 (Foc4) of Fusarium wilt of banana, and the tested banana variety is Brasilia Cavendish (AAA).

[0041] 1.2 Host bacteria and plasmid vector

[0042]The host bacterium was Escherichia coli DH5α, the cloning vector was pMD18-T vector, and the gene knockout vector was pUC19-HPH (reformed by our laboratory on the basis of pUC19 plasmid, that is, inserted between the Kpn I and BamHI sites on pUC19 TrpC+HPH fragment), the gene complement carrier is pCTZN (reformed by our laboratory on the basis of the pCT74 plasmid, that is, replacing the SGFP and hph genes on pCT74 with the bleomycin (Zeocin) gene), also in the patent "201910986131.4 , The application of the gene FoPDCD5 in regulating the pathogenicity of Fusarium wilt of banana".

[0043] 2. Experimental method

[0044] 2.1 Amplification of upstream and downstream homologous fragments of FoPao gene of Fusarium wilt of bana...

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Abstract

The invention discloses application of a gene FoPao in regulation and control of pathogenicity of banana fusarium wilt, and belongs to the field of plant genetic engineering. The application comprisesthe following steps: constructing a gene knockout vector, and introducing the vector into a banana fusarium wilt protoplast; and knocking out the gene from banana fusarium wilt by using a homologousrecombination method to obtain a knockout mutant delta FoPao. The gene supplementing vector is constructed, the gene supplementing vector is introduced into the delta FoPao protoplast, and the gene issupplemented back to the knockout mutant by using a random insertion method to obtain the supplemented mutant delta FoPao-com. Pathogenicity tests show that the deletion of the gene FoPao significantly increases the toxicity of banana fusarium wilt; and after the gene is supplemented, the toxicity of the supplemented mutant is consistent with the wild type of the banana fusarium wilt. The invention proves that the FoPao is related to pathogenicity of banana fusarium wilt. The research is helpful to deeply clarify the pathogenic molecular mechanism of banana fusarium wilt.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to the application of a gene FoPao in regulating the pathogenicity of Fusarium wilt of banana. Background technique [0002] Banana Fusarium wilt, also known as Panama disease, is a devastating soil-borne vascular disease caused by Fusarium oxysporumf.sp.cubense (Foc). ". Because the disease spreads rapidly, it is difficult to cure it, which seriously affects the development of the banana industry. [0003] In the secretome study of Fusarium wilt, we identified a predicted polyamine oxidase (N1-acetylpolyamine oxidase, predicted, PAO) containing amino oxidase (Amino oxidase) domain. Polyamine oxidase is an important enzyme in the polyamine metabolic pathway of fungi, which can oxidize acetylspermidine to spermidine, or oxidize acetylspermine to spermine; polyamine metabolism helps maintain the balance of polyamines in organisms dynamic equilibrium. [0004] At present, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/06C12N15/80C12N15/90C12Q1/18C12R1/77
CPCC12N9/0032C12N15/80C12N15/902C12Q1/18C12Y105/03011G01N2333/37
Inventor 聂燕芳李云锋颜瑞李华平
Owner SOUTH CHINA AGRI UNIV
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