CN-122012603-A - Gene GhTLP b for improving verticillium wilt resistance of cotton and application thereof

Abstract

The invention relates to application of a cotton sweet protein gene GhTLP b in improving verticillium resistance of cotton. The invention clones GhTLP b gene from disease-resistant material ZZM, utilizes agrobacterium-mediated transformation technology to obtain transgenic cotton strain with over-expression and gene silencing of the gene, and proves that GhTLP b over-expression strain enhances the resistance of cotton to verticillium wilt, and is characterized by delayed onset and reduced disease index, ghTLP b silencing strain causes the cotton to be susceptible, and is characterized by reduced active oxygen burst and reduced accumulation of lignin and callose. The invention identifies and verifies the important resistance gene of cotton for resisting verticillium wilt, and provides effective gene resources for genetic breeding.

Inventors

• LI ZHANSHUAI
• LIU YUANYUAN
• WEI FENG
• PENG JUN
• HE JIE
• ZHANG XINJIE
• YANG LU
• WANG YUAN

Assignees

• 三亚中国农业科学院国家南繁研究院

Dates

Publication Date

20260512

Application Date

20260410

Claims (10)

1. The application of the GhTLP1b gene in improving the verticillium wilt resistance of cotton is characterized in that the nucleotide sequence of the GhTLP b gene is shown as SEQ ID NO.1, and the improvement of the resistance of cotton to verticillium wilt is shown as the enhancement of active oxygen burst, lignin accumulation and callose accumulation.
2. 2. The use according to claim 1, wherein the sequence of the protein coding region of the GhTLP b gene is shown in SEQ ID No.2 and the amino acid sequence of the protein is shown in SEQ ID No. 3.
3. 3. The use according to claim 1, wherein the use is to increase the resistance of cotton to verticillium by increasing the expression level or protein activity of GhTLP b gene in cotton.
4. 4. The method of claim 3, wherein the enhanced resistance to verticillium in cotton is further characterized by delayed onset and reduced disease index.
5. 5. The use of claim 1, further comprising attenuating resistance to verticillium by silencing or reducing GhTLP b gene expression or protein activity in cotton.
6. 6. The use according to claim 5, wherein the impaired cotton resistance to verticillium is manifested by reduced bursts of active oxygen, reduced lignin accumulation, reduced callose accumulation.
7. The application of the GhTLP1b gene in cotton verticillium wilt resistance genetic breeding is characterized in that the nucleotide sequence of the GhTLP b gene is shown as SEQ ID NO.1, and the improvement of the cotton resistance to verticillium wilt is shown as the enhancement of active oxygen burst, lignin accumulation and callose accumulation.
8. 8. A biological material for improving verticillium wilt resistance of cotton is characterized in that the biological material is an over-expression strain obtained by over-expressing GhTLP b genes in cotton, the nucleotide sequence of the GhTLP b genes is shown as SEQ ID NO.1, and the improvement of the verticillium resistance of cotton is shown by the enhancement of active oxygen burst, the increase of lignin accumulation and the increase of callose accumulation.
9. 9. A method for enhancing verticillium wilt resistance of cotton is characterized by comprising the step of enhancing expression quantity or protein activity of GhTLP b gene in receptor cotton, wherein the nucleotide sequence of GhTLP b gene is shown as SEQ ID NO.1, and the enhancement of verticillium resistance of cotton is shown as enhancement of active oxygen burst, lignin accumulation and callose accumulation.
10. 10. The method of claim 9, wherein increasing the expression level or protein activity of GhTLP b gene in the recipient cotton is achieved by introducing into the recipient cotton and over-expressing an expression vector comprising the coding region sequence set forth in SEQ ID No. 2.

Description

Gene GhTLP b for improving verticillium wilt resistance of cotton and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a gene GhTLP b for improving verticillium wilt resistance of cotton and application of the gene GhTLP b in improving verticillium wilt resistance of cotton. Background Cotton is an important commercial crop whose yield and fiber quality are often affected by a variety of diseases, with verticillium dahliae (Verticillium dahliae) causing verticillium wilt being a serious soil-borne vascular bundle disease. After the pathogenic bacteria infects from root systems, the plant vascular bundles are blocked, so that leaves are yellow, wilting and falling off, the plants die when serious, and the yield of cotton is reduced by 10% -30% all the year round. At present, the disease lacks a high-efficiency stable control method, and has long pathogenic bacteria soil survival period and wide host range, so that the effects of measures such as rotation and the like are limited. Therefore, breeding disease-resistant varieties becomes a strategy for controlling the disease to be durable and effective. Plants have developed a complex immune system in long term evolution, where pathogen-associated Proteins (PRs) play a key role in disease resistance. Sweet proteins (TLPs) belong to the PR-5 family, are widely present in plants and have been reported to be involved in responses to fungal, bacterial and abiotic stresses. TLPs typically have a β -sheet structure, which inhibits their growth by disrupting the pathogenic cell membrane or directly interacting with the pathogenic bacteria. Several TLP members have been found in cotton, but their specific functions and mechanisms in verticillium resistance are not yet defined, especially the lack of key genes with defined disease resistance effects that can be used in molecular breeding. The current cotton verticillium wilt resistance breeding mainly depends on traditional hybridization and phenotype selection, and has the problems of long period, low efficiency, complex resistance genetic basis and the like. Although the related genes of disease resistance are reported, the key genes which are proved by functions and have obvious disease resistance effect are few, and the breeding development of disease resistance molecules is limited. Therefore, the method has important significance in improving the verticillium wilt resistance of cotton and guaranteeing the production safety by digging the cotton endogenous gene with definite disease resistance, clarifying the mechanism and developing the application of the cotton endogenous gene in disease resistance breeding. Disclosure of Invention The invention aims to provide a cotton verticillium wilt resistant material ZZM and application of a disease resistance gene GhTLP b carried by the cotton verticillium wilt resistant material in enhancing cotton verticillium wilt resistance, and provides effective germplasm resources for cultivating a cotton upland variety resistant to verticillium wilt. The invention identifies and clones a resistance gene GhTLP b for enhancing verticillium wilt of cotton, proves that GhTLP b up-regulates expression after verticillium dahliae induction, and the gene can enhance the resistance of cotton to verticillium wilt after over-expression in cotton, and plant susceptibility is improved after silencing the gene. In order to achieve the aim, the invention provides a gene GhTLP b for regulating and controlling verticillium wilt resistance of cotton, and the nucleotide sequence of the gene is shown as SEQ ID NO. 1. The invention also provides the coding protein of the gene, the coding region sequence (CDS) of the coding protein is shown as SEQ ID NO.2, and the corresponding amino acid sequence is shown as SEQ ID NO. 3. The invention also provides an application of GhTLP b gene in improving verticillium wilt resistance of cotton, and a nucleotide sequence of GhTLP b gene is shown as SEQ ID NO. 1. Preferably, the use is to enhance the resistance of cotton to verticillium by increasing the expression level or protein activity of GhTLP b gene in cotton. Preferably, the enhanced cotton resistance to verticillium is manifested by delayed onset and reduced disease index. Preferably, the use further comprises attenuating resistance to verticillium in cotton by silencing or reducing the amount of GhTLP b gene expression or protein activity in cotton. Preferably, the impaired cotton resistance to verticillium is manifested by reduced bursts of active oxygen, reduced lignin accumulation, and reduced callose accumulation. The invention also provides a biological material for improving verticillium wilt resistance of cotton, which is an over-expression strain obtained by GhTLP b genes in cotton. The invention also provides an application of GhTLP b gene in cotton verticillium wilt resistant genetic breeding. The invention also p