Application of gene GhSWEET42 in prevention and treatment of cotton verticillium wilt

A cotton Verticillium wilt and anti-Verticillium wilt technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of lack of cotton germplasm resources and limited means of prevention and control of Verticillium wilt, so as to improve resistance and reduce Effect of Glucose Content

Active Publication Date: 2021-11-12
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the prevention and control methods for Verticillium wilt are very limited at present, and there is a lack of cotton germplasm resources with high resistance to Verticillium wilt

Method used

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  • Application of gene GhSWEET42 in prevention and treatment of cotton verticillium wilt
  • Application of gene GhSWEET42 in prevention and treatment of cotton verticillium wilt
  • Application of gene GhSWEET42 in prevention and treatment of cotton verticillium wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1. Materials

[0051] 1. Experimental materials and strains

[0052] Plant materials required for the experiment: cotton is the selfed seed of Zhongzhimian No. 2 in upland cotton; strains required for the experiment: Verticillium dahliae strain V592 with GFP, Verticillium dahliae strain V592 preserved in our laboratory.

[0053] 2. V592 culture and V592 infection

[0054] To determine the colonization process of Verticillium dahliae strain V592 in cotton, Verticillium dahliae with green fluorescent protein (GFP) was used to infect cotton and the fluorescence was observed under a confocal microscope. details as follows:

[0055] Take 100 μL of the spore liquid of Verticillium dahliae V592 with GFP stored at -80°C, inoculate it on the potato dextrose agar medium, cultivate it at a constant temperature and dark for one week at 25°C, cut the mycelium pieces the size of your thumb into pieces, and inoculate them on the In the culture medium, shake culture in a constant te...

Embodiment 2

[0068] 1. Association analysis of genes

[0069]The protein sequence of the grape SWEET gene family was downloaded from the Grape Genome website, and the protein sequence of the Arabidopsis SWEET gene family was downloaded from the TAIR database. The protein sequence of cotton (G. upland) was downloaded from the CottonGen website. The homology of the protein sequences of SWEET upland cotton, grapevine and Arabidopsis was compared, and then the phylogenetic tree was constructed by the neighbor-joining method, and the phylogenetic tree was constructed by MEGA (version 7.0).

[0070] Utilizing the protein sequences of SWEET family members of upland cotton, Arabidopsis SWEET family members and grape SWEET family members, MEGA7 software was used for evolutionary analysis, combined with previous literature research, the evolutionary tree was divided into four subgroups ( image 3 A). Because Verticillium dahliae infection can cause significant up-regulation of glucose content, the...

Embodiment 3

[0085] Arabidopsis overexpressing GhSWEET42 reduces resistance to Verticillium wilt

[0086] 1. Heterologous overexpression in Arabidopsis

[0087] (1) Take out the Agrobacterium liquid containing the recombinant plasmid (GhSWEET42-pCambia2300) from -80°C, activate the bacterial liquid, and use the resuspension liquid to adjust the OD of the bacterial liquid to 600 Adjust to 1.2, add the surfactant containing 0.02% silwet-77 into the bacterial liquid, and mix well.

[0088] (2) Two days before the blooming period of Arabidopsis thaliana, cut off the fruit pods, soak the cut Arabidopsis thaliana in the prepared bacterial solution for 60 seconds, then treat it in the dark for 24 hours, and then put it in the plant greenhouse normal growth.

[0089] (3) After Arabidopsis recovers to normal growth, carry out secondary infection.

[0090] (4) After the fruit pods of Arabidopsis thaliana matured, the seeds were harvested, placed in a centrifuge tube added with a desiccant, and th...

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Abstract

The invention relates to the technical field of prevention and treatment of cotton diseases, in particular to application of a gene GhSWEET42 in prevention and treatment of cotton verticillium wilt, and the nucleotide sequence of the gene GhSWEET42 is as shown in SEQ ID NO: 1. The inventor finds that the expression condition of the gene GhSWEET42 is closely related to the infection process of the cotton verticillium wilt through verification in multiple aspects. When the verticillium dahliae infects the cotton root, the glucose content is obviously increased in the colonization process of the verticillium dahliae, and the expression quantity of the gene GhSWEET42 is obviously up-regulated when the verticillium dahliae invades; and subcellular localization and yeast heterologous complementation experiments prove that the gene GhSWEET42 encoding protein is a glucose transporter localized on plasma membranes. The gene GhSWEET42, which is a key gene of verticillium dahliae in cotton infection, is further obtained through an overexpression gene GhSWEET42 and a silence gene GhSWEET42.

Description

technical field [0001] The invention relates to the technical field of cotton disease control, in particular to the application of gene GhSWEET42 in the control of cotton verticillium wilt. Background technique [0002] Verticillium wilt is a soil-borne fungal disease caused by Verticillium dahliae, which can infect many economically important crops. The growth cycle of Verticillium dahliae begins with spore germination, and the produced hyphae directly penetrate the roots of the plant and settle in the xylem vascular bundles. They can survive in the xylem of the plant for a long time without causing obvious disease symptoms. . When symptoms occur, they usually include wilting, wilting, yellowing, and abscission of leaves, stunted plant growth, decreased photosynthesis, and discoloration of vascular tissues, and the type of symptoms can vary depending on the host plant. Symptoms such as leaf wilting and yellowing are closely related to fungal conidia and mycelium produced ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/60C12Q1/6895
CPCC07K14/415C12N15/8282C12Q1/6895C12Q2600/158C12Q2600/13
Inventor 张飞杨代刚马雄风李威孙梦玺王星星任中英赵俊杰裴小雨刘艳改贺昆仑周晓箭张文生郭金峰
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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