Application of Gene ghsinas in Controlling Cotton Verticillium Wilt

A cotton Verticillium wilt and anti-Verticillium wilt technology, applied in the field of plant disease control, can solve problems such as unknown functions and applications

Active Publication Date: 2022-12-02
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 function and application of the SINA E3 ligase associated with Verticillium wilt resistance remain unknown

Method used

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  • Application of Gene ghsinas in Controlling Cotton Verticillium Wilt
  • Application of Gene ghsinas in Controlling Cotton Verticillium Wilt
  • Application of Gene ghsinas in Controlling Cotton Verticillium Wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Bioinformatics Analysis

[0093] In order to study the role of cotton SINA ubiquitin ligases, a total of 12 Asian cotton, 12 Raymond cotton and 24 upland cotton SINA genes were identified in the cotton genome database. Among them, 3 cotton varieties of Upland cotton, Asian cotton and Raymond cotton were obtained from the CottonFGD database (http: / / www.cottonfgd.org / ). Afterwards, the upland cotton SINA gene was cloned, and the coding sequence of GhSINA8 obtained by sequencing was modified on the basis of its initial annotation. These SINAs encode two typical conserved domains, a RING finger domain (at the amino-terminus)) and a SINA domain (at the carboxy-terminus) ( figure 1 A)). In order to study the sequence of the SINA homology domain of upland cotton and the conserved frequency of each residue, the sequence alignment was performed and the sequence characteristics of the SINA protein were obtained, such as figure 1 As shown in B, the sequence identification map s...

Embodiment 2

[0097] Expression analysis of GhSINA gene in response to Verticillium wilt

[0098] 24 hours after GFP-labeled Verticillium dahliae infected upland cotton, it was detected that the conidia on the roots of cotton seedlings had germinated and the hyphae of Verticillium dahliae had grown and extended. In order to determine whether the expression levels of GhSINA genes changed 24 hours after Verticillium dahliae infection, their transcription levels in the roots of an upland cotton resistant variety (Zhongzhimian No. 2) were studied, and the primers involved are shown in Table 1. Among them, GhSINA7, GhSINA8, and GhSINA9 exist in subgroup A chromosome and subgroup D chromosome respectively, but the sequences of their homologous gene pairs are highly similar, and it is difficult to distinguish these homologous gene pairs by real-time fluorescent PCR (qRT-PCR). , so they are amplified together. figure 2 The expression level of SINA gene in Zhimian No. 2, a resistant variety, was i...

Embodiment 3

[0103] 1. Yeast two-hybrid experiment

[0104] Yeast two-hybrid screening was constructed according to the instructions of the Matchmaker Gold yeast two-hybrid system (Clontech, Palo Alto, CA, USA). The full-length cDNAs of GhSINAs (see Table 2 for specific primers) were cloned into the bait vector pGBDK7 and the prey vector pGADT7, respectively. The constructed recombinant plasmid was co-transformed into yeast Y2HGold, and cultured on SD / -Leu / -TrpDO (DDO) medium at 30°C for 3 days. Pick independent colonies of the same size and transfer them to SD / -Leu / Trp / -Ade / -His DO (QDO) medium containing X-α-Gal (Clontech), and evaluate the GhSINA protein by whether the colonies turn blue. interaction between.

[0105] Table 2 GhSINAs primers for different vectors

[0106]

[0107]SINA proteins perform biological functions by forming homodimers or heterodimers. In order to determine whether these three candidate GhSINAs proteins have the ability to form homo- or hetero-dimers, the...

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Abstract

The invention relates to the technical field of plant disease control, in particular to the application of gene GhSINAs in the control of cotton verticillium wilt. The invention provides the application of gene GhSINAs in the prevention and treatment of cotton verticillium wilt, said gene GhSINAs comprises any one or more in gene GhSINA7, gene GhSINA8 and gene GhSINA9; The nucleic acid sequence of gene GhSINA7 is as SEQ ID NO:1 and / or shown in SEQ ID NO:2; the nucleic acid sequence of gene GhSINA8 is shown in SEQ ID NO:3 and / or SEQ ID NO:4; the nucleic acid sequence of gene GhSINA9 is shown in SEQ ID NO:5 and / or SEQ ID NO:6 shown. The present invention found in the research that among these three genes, each overexpressed transgenic line has improved resistance to Verticillium wilt, but the silencing of each gene has suppressed the defense ability to pathogen infection, indicating that GhSINA7, GhSINA8 and GhSINA9 play a positive regulatory role in cotton defense against Verticillium dahliae.

Description

technical field [0001] The invention relates to the technical field of plant disease control, in particular to the application of gene GhSINAs in the control of cotton verticillium wilt. Background technique [0002] Verticillium dahliae is a devastating soil-borne fungal pathogen that causes Verticillium wilt, which affects the growth and development of more than 200 plant species, including cotton, an important agricultural economic crop. As we all know, Verticillium dahliae is a very difficult to control pathogen, which has strong infection pathogenicity and strong survival ability in soil, so that Verticillium dahliae can penetrate the main root xylem vessel and vascular vascular tissue, which severely clogs the plant's vessels, eventually leading to stunting and wilting of the plant. In addition, the pathogen has a wide host range and can survive in soil for many years without a host. Breeding of disease-resistant varieties is an effective and practical strategy to co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/61C12N15/82C12N15/11C12Q1/6895A01H5/00A01H6/60
CPCC12N9/93C12N15/8282C12Q1/6895C12Y603/02019C12Q2600/13C12Q2600/158
Inventor 任中英杨代刚马雄风李威王星星赵俊杰裴小雨刘艳改贺昆仑张飞周晓箭张文生郭金峰
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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