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CN-121992035-A - GhHSL1 gene and application thereof in regulation and control of upland cotton verticillium wilt resistance

CN121992035ACN 121992035 ACN121992035 ACN 121992035ACN-121992035-A

Abstract

The invention relates to the field of biotechnology application, and discloses GhHSL genes and application thereof in regulating and controlling verticillium wilt resistance of upland cotton. The nucleotide sequence of GhHSL gene upland cotton genetic standard line TM-1 is shown as SEQ ID NO. 1, and the amino acid sequence is shown as SEQ ID NO. 2. The gene is obviously up-regulated by the induction of verticillium wilt. The invention further utilizes virus-induced gene silencing technology and transgenic over-expression technology to perform preliminary study on the function of cotton GhHSL gene in verticillium wilt resistance of cotton, and results show that GhHSL gene in upland cotton is silenced to obviously reduce verticillium wilt resistance of cotton plants, ghHSL gene is over-expressed in upland cotton to obviously improve verticillium wilt resistance of cotton, and GhHSL1 gene forward regulation and control on verticillium dahliae resistance of upland cotton is determined, and the invention firstly reveals the application of GhHSL1 gene in regulating verticillium wilt resistance of upland cotton and provides theoretical guidance and genetic engineering for improving verticillium wilt resistance of cotton through molecular design breeding or genetic engineering.

Inventors

  • HU YAN
  • XIONG HUIFANG
  • LI XIAORAN

Assignees

  • 浙江大学

Dates

Publication Date
20260508
Application Date
20260408

Claims (8)

  1. The application of the GhHSL1 gene in regulating and controlling the verticillium wilt resistance of upland cotton is characterized in that the nucleotide sequence of the GhHSL gene is shown as SEQ ID NO.1, and the encoded protein sequence is shown as SEQ ID NO. 2.
  2. 2. The use of claim 1, wherein overexpression of GhHSL gene increases upland cotton verticillium resistance and silencing GhHSL1 gene decreases upland cotton verticillium resistance.
  3. 3. The use according to claim 1, wherein the over-expression GhHSL gene can improve the verticillium wilt resistance of upland cotton by constructing an expression cassette, a recombinant expression vector, a recombinant host cell or a recombinant bacterium containing GhHSL gene and transferring the expression cassette, the recombinant vector, the recombinant host cell or the recombinant bacterium into upland cotton, thereby improving the verticillium wilt resistance of upland cotton.
  4. 4. The use of claim 1, wherein the silencing GhHSL gene reduces upland cotton verticillium resistance by amplifying the primers to obtain a VIGS fragment, and performing virus-induced gene silencing on upland cotton, thereby reducing upland cotton GhHSL1 gene expression to reduce upland cotton verticillium resistance.
  5. 5. The use according to claim 1, wherein the primers comprise an upstream primer VIGS-F and a downstream primer VIGS-R, the nucleotide sequence of the upstream primer is shown in SEQ ID No.5, and the nucleotide sequence of the downstream primer is shown in SEQ ID No. 6.
  6. 6. The use according to any one of claims 1 to 5, wherein the verticillium wilt is a plant disease caused by verticillium dahliae Verticillium dahliae.
  7. 7. A method of increasing verticillium resistance in a plant comprising: a) Constructing an over-expression recombinant plant expression vector containing GhHSL genes; b) Transforming plants with an over-expression recombinant plant expression vector containing GhHSL genes to enable GhHSL genes to be over-expressed in the plants, so that the verticillium resistance of the plants is improved; Wherein the plant is upland cotton, and the verticillium wilt is a plant disease caused by verticillium dahliae Verticillium dahliae.
  8. 8. A method of growing a verticillium wilt resistant plant variety comprising: a) Constructing an over-expression recombinant plant expression vector containing GhHSL genes; b) Transforming plants with an over-expression plant expression vector containing GhHSL genes to enable GhHSL genes to be over-expressed in the plants, obtaining transgenic positive plants and screening to obtain plant varieties with improved verticillium resistance; Wherein the plant is upland cotton, and the verticillium wilt is a plant disease caused by verticillium dahliae Verticillium dahliae.

Description

GhHSL1 gene and application thereof in regulation and control of upland cotton verticillium wilt resistance Technical Field The invention belongs to the field of biotechnology application, and particularly relates to GhHSL gene and application thereof in regulating and controlling upland cotton verticillium wilt resistance. Background Cotton (Gossypium spp.) has found wide use in agriculture, textile industry, and medical and health fields as one of the most prominent commercial crops worldwide. China is the largest cotton producing country worldwide, but faces a serious disease called "cotton cancer", cotton verticillium wilt. The disease can cause yellowing and wilting of cotton plant leaves, not only greatly reduces cotton yield, but also seriously damages cotton fiber quality, and becomes a main factor affecting cotton production. Cotton verticillium wilt (Verticillium wilt) is a fungal soil-borne disease, pathogenic bacteria are verticillium dahliae (Verticillium dahliae Kleb), and the pathogenic bacteria can cause systemic disease by infecting cotton vascular bundles, and the cotton verticillium dahliae is characterized by wide distribution, serious hazard and long survival time, and has become a main obstacle for the production of cotton which is a worldwide hazard. The cotton has huge genome, complex genetic background, deficient verticillium wilt resistance gene resource and complex verticillium wilt resistance pathogenesis, and the factors restrict the research of verticillium wilt resistance and the molecular breeding progress of verticillium wilt resistance of cotton. The cloning and functional identification of the cotton verticillium wilt resistance gene are developed, and the research on the molecular mechanism of the interaction of cotton and verticillium wilt resistance bacteria is an important subject of cotton verticillium wilt resistance breeding research. However, the pathogen causing the disease lives in soil layers of 40 cm, and the variation is frequent, so that the traditional bactericides and chemical agents are difficult to prevent and treat. And the host of the plant is up to more than 120 plants, and the short-term effect of the traditional crop rotation disease-resistant mode is also not ideal. Therefore, planting verticillium resistant varieties is the most effective method of controlling the disease. However, the disease resistance of cotton varieties in China can only reach the level of disease resistance, so that the cultivation of good varieties with high verticillium wilt resistance is still a bottleneck to be solved in cotton variety breeding. Research on the pathogenic mechanism of verticillium wilt and the disease resistance mechanism of cotton, and the transfer of resistance characters into the existing excellent cultivars by using molecular markers and genetic engineering technology is the current direction of verticillium wilt-resistant breeding. Disclosure of Invention The invention aims at overcoming the defects of the prior art and provides GhHSL gene and potential and application thereof in regulating and controlling verticillium wilt resistance of upland cotton and cultivating new cotton germplasm with verticillium wilt resistance. In order to achieve the above purpose, the present invention proposes the following technical scheme: In a first aspect, the invention provides GhHSL genes which are separated from upland cotton and are related to verticillium wilt resistance regulation, wherein the nucleotide sequence of the GhHSL genes is shown as SEQ ID NO. 1, and the coded protein sequence of the GhHSL genes is shown as SEQ ID NO. 2. A GhHSL gene related to cotton verticillium wilt resistance has a nucleotide sequence shown in SEQ ID NO.1 in upland cotton standard line TM-1. The expression level of the gene is positively correlated with the verticillium wilt resistance of cotton, and the over-expression GhHSL gene in upland cotton can obviously reduce the sensitivity of the upland cotton to verticillium wilt, reduce the disease index, lighten the blackness degree of vascular bundles and enhance the content of gossypol, and the reduction of GhHSL gene expression in upland cotton can obviously enhance the sensitivity of the upland cotton to verticillium wilt, improve the disease index, increase the blackness degree of vascular bundles and reduce the content of gossypol. In a second aspect, the invention provides an application of GhHSL gene in enhancing upland cotton verticillium wilt resistance, wherein expression of GhHSL gene is improved in upland cotton, and upland cotton verticillium wilt resistance is enhanced. The GhHSL gene shown as the over-expressed SEQ ID NO. 1 is applied to enhancing the verticillium wilt resistance of upland cotton or cultivating new cotton varieties resistant to verticillium wilt. In a third aspect, the invention provides the application of the biological material for improving GhHSL gene expression in enhancing upland cotton verticil