CN-121975830-A - Application of cotton GbABH gene 132 in verticillium wilt resistance of plants

Abstract

The invention discloses a cotton gene GbABH and application thereof in plant verticillium wilt resistance, the nucleotide sequence of the gene is shown as SEQ ID No.1, and the coded amino acid sequence is shown as SEQ ID No. 2. The cotton plant with the gene silenced by the VIGS technology has increased susceptibility to verticillium wilt, and the cotton gene GbABH132,132 is introduced into Arabidopsis thaliana to obtain transgenic plant with stronger verticillium wilt resistance than wild Arabidopsis thaliana. Therefore, the invention has important significance in breeding and researching the improvement of the verticillium wilt resistance of cotton and can be applied to breeding of disease-resistant cotton varieties.

Inventors

• LI PENGTAO
• LU QUANWEI
• PENG RENHAI
• LIU XIAOYAN
• WANG WENKUI
• Cheng Mingzheng
• GE QUN
• YANG RUI
• GONG JUWU
• WEI YANGYANG
• LIU YULING

Assignees

• 安阳工学院

Dates

Publication Date

20260505

Application Date

20260305

Claims (5)

1. 1. A cotton gene GbABH132,132 is characterized in that the nucleotide sequence of the gene is shown as SEQ ID No.1, and the amino acid sequence encoded by the gene is shown as SEQ ID No. 2.
2. 2. The use of the cotton gene GbABH132,132 as claimed in claim 1 for plant verticillium wilt resistance.
3. 3. The use of cotton gene GbABH132,132 as claimed in claim 2 for the resistance of plants to verticillium wilt, wherein said plants are cotton or arabidopsis.
4. 4. The use of cotton gene GbABH132,132 as claimed in claim 2 for plant verticillium wilt resistance, wherein verticillium wilt resistance of plants is enhanced by over-expression of said GbABH132,132 gene in plants.
5. 5. A method for obtaining a verticillium wilt resistant crop comprising the steps of: (1) Obtaining GbABH gene as defined in claim 1 and connecting with expression vector plasmid to construct recombinant vector containing GbABH 132; (2) Transforming the recombinant vector containing GbABH132,132 into competent cells of agrobacterium tumefaciens by a freeze thawing method, and screening to obtain positive transformants; (3) Transfecting bacterial liquid containing positive transformant into wild crop, harvesting T0 generation after the crop is mature, screening T0 generation to obtain transgenic crop with verticillium resistance; (4) And selecting the homozygote of the T3 generation GbABH gene with stable neutrality of the selfing offspring for reservation to obtain the verticillium wilt-resistant crop.

Description

Application of cotton GbABH gene 132 in verticillium wilt resistance of plants Technical Field The invention particularly relates to application of cotton GbABH gene 132 in verticillium wilt resistance of plants, and belongs to the technical field of cotton genetic engineering. Background Verticillium wilt of plants is a typical soil-borne fungal vascular bundle disease caused by verticillium dahliae infection. In China, verticillium dahliae is mainly used as a dominant pathogen, more than 400 plants can be caused to generate diseases, and a large amount of crop losses can be caused each year. Verticillium wilt is extremely difficult to control, mainly because verticillium dahliae is a xylem colonizing pathogen, and no effective treatment method can cure infected plants at present. Another reason is that verticillium dahliae produces a microhard core, i.e. a darkened resting structure, that is able to survive for decades in the soil. Morphology and physiological symptoms of cotton infected with verticillium dahliae include leaf wilting, hair Huang Juanqu, and vascular bundle browning. The plant-growth-promoting and plant-growth-promoting agent has multiple pathogenic bacteria physiology, rapid variation, can survive for a plurality of years in soil, can germinate repeatedly, is difficult to thoroughly control, and seriously damages the production and development of crops. Cotton planting areas in China are wide, and the yield is nearly one fourth of the global total yield. However, the cotton loss caused by verticillium accounts for 32.49% of the total loss of other different diseases, and the cotton verticillium prevention and control of cotton verticillium are extremely difficult due to single cotton planting structure, lack of cotton resistance resources, high verticillium mutation speed and the like in China. The current selection of cotton with excellent verticillium wilt resistance is the most economical and effective strategy for solving the dilemma. The Alpha/beta hydrolase superfamily (ABH) family is a widely distributed and functionally plastic hydrolase, known for its diverse biochemical activity in three areas of life. ABHs are generally associated with functions involved in cell metabolite breakdown and recycling, external nutrient processing, and detoxification of exogenous substances. In addition ABHs plays a key regulatory role in metabolism, such as carboxypeptidase II is involved in regulating protein longevity, function and turnover, and phospholipase produces second messengers such as diacylglycerol and phosphatidic acid that regulate cellular function and respond to environmental stimuli. The ABH family also supports a variety of unique catalytic functions for defense and hormonal regulation. Cotton related studies have reported that salicylic acid binding protein 2 (SABP 2) can regulate the response to salicylic acid, and is involved in plant immune response regulation. Gibberellin receptors are members of the ABH family, and GID1 as a receptor binds gibberellin, activating a plant immune response. In the research of the invention, a cotton gene GbABH132,132 is cloned and identified, and the sequence structure, functional analysis and expression pattern analysis show that the cotton gene 3835,132 can be expressed in a large amount at the root when being infected by verticillium, and then a transgenic plant is created by using a gene silencing and over-expression method, thereby verifying the important role of the cotton gene 3835,132 in resisting verticillium. Disclosure of Invention The invention aims at identifying cotton gene GbABH132,132 and researching the application of the gene in improving verticillium wilt resistance of plants. In a first aspect, the invention provides a cotton gene GbABH132,132, the nucleotide sequence of which is shown in SEQ ID No.1, and the encoded amino acid sequence is shown in SEQ ID No. 2. In a second aspect, the invention provides the use of cotton gene GbABH132,132 in plant verticillium wilt resistance. Further, the plant is cotton or arabidopsis thaliana. Further, the application is to introduce a cotton gene GbABH to Arabidopsis thaliana to obtain a transgenic plant with stronger verticillium resistance compared with wild Arabidopsis thaliana. The invention identifies a cotton gene GbABH132,132 which can improve plant disease resistance for the first time. Silencing the expression level of the gene in the medium cotton plant No. 36, liu Haijian osmotic MBI8255 increases the susceptibility of cotton to verticillium wilt, while increasing verticillium wilt resistance in Arabidopsis is achieved by an excessive amount. In a third aspect, the invention also provides a method of obtaining a verticillium wilt resistant crop comprising the steps of: (1) Obtaining GbABH gene as defined in claim 1 and connecting with expression vector plasmid to construct recombinant vector containing GbABH 132; (2) Transforming the recombinant vector containing GbABH132,132