CN-122012501-A - Application of RWTD gene in prevention and control of sugarcane smut

Abstract

The invention discloses an application of RWTD gene in prevention and control of sugarcane smut, wherein the gene is a key factor for regulating sexual coordination of pathogenic bacteria and development of pathogenic symptoms (black whip). The functional verification proves that the pathogenic bacteria can not complete the whole life history after RWTD gene deletion, and even if the pathogenic bacteria can infect a host, the pathogenic bacteria can not induce plants to form black whips and produce winter spores, thereby cutting off disease transmission chains. Based on the gene, RWTD gene, encoding protein or related regulatory sequences thereof are used as control targets, various application approaches including RNA interference, gene editing, chemical inhibition and disease-resistant breeding are provided, and a core molecular foundation and a brand new solution are provided for developing a high-efficiency, accurate and environment-friendly novel sugarcane smut prevention and control technology.

Inventors

• LU PAN
• TAN SHAOFENG
• Wu Nianchen
• ZHOU CHUNLING
• LIANG MI
• LI XIAOYI
• CHEN LIRONG

Assignees

• 广西大学

Dates

Publication Date

20260512

Application Date

20260226

Claims (10)

1. 1. A method for controlling sugarcane smut, which comprises inhibiting the function and/or expression of RWTD gene in alternaria alternata (Sporisorium scitamineum), wherein the nucleotide sequence of RWTD gene is shown as SEQ ID No.1 or encodes a protein with the same function.
2. 2. The method of claim 1, wherein the inhibition is achieved by administering an RNA interference molecule, antisense nucleic acid, or gene editing system for the RWTD gene to sugarcane, a sugarcane growing environment, or a septoria Saccharopolyspora.
3. 3. The method of claim 1, wherein the inhibition is achieved by applying a compound capable of inhibiting RWTD protein activity to sugarcane, a sugarcane growing environment, or a septoria Saccharum sinensis Roxb.
4. 4. A method according to any one of claims 1 to 3, wherein the inhibition results in inhibition of black whip formation and/or winter spore development following infection of sugarcane by the septoria Saccharopolyspora.
5. 5. The method of claim 4, wherein the inhibition results in the ability of the alternaria alternata to infect sugarcane but not to induce black whip.
6. Application of RWTD1 gene or coded protein thereof in preventing and treating sugarcane smut, wherein the nucleotide sequence of RWTD gene is shown as SEQ ID No.1 or codes protein with the same function.
7. 7. A kit for preventing and treating sugarcane smut is characterized by comprising an active ingredient for inhibiting RWTD gene functions and/or expression in the sugarcane black powder, wherein the active ingredient is selected from an RNA interference molecule targeting the RWTD1 gene, an antisense nucleic acid, a gene editing system or a compound capable of inhibiting RWTD protein activity.
8. 8. A recombinant microorganism is characterized by comprising an expression vector capable of expressing an RNA interference molecule or antisense nucleic acid of a target Saccharopolyspora sinensis RWTD gene or an expression vector capable of expressing a protein inhibiting RWTD protein activity, wherein the nucleotide sequence of the RWTD gene is shown as SEQ ID No.1 or encodes a protein with the same function, and the recombinant microorganism is a bacterium or a fungus.
9. 9. A molecular marker for identifying or monitoring the effect of preventing and controlling sugarcane smut, characterized in that the molecular marker is the expression level or functional state of the gene RWTD of the fusarium graminearum, wherein the down-regulation or functional inactivation of RWTD1 gene indicates that prevention and control are effective.
10. 10. A method of growing an anti-smut sugarcane plant, the method comprising introducing into sugarcane a construct capable of expressing a dsRNA, siRNA or shRNA of a RWTD gene, thereby producing in the sugarcane an interfering RNA targeting the fusarium graminearum RWTD gene.

Description

Application of RWTD gene in prevention and control of sugarcane smut Technical Field The invention belongs to the technical field of plant disease prevention and control, and particularly relates to application of RWTD gene in prevention and control of sugarcane smut. Background Sugar cane (Saccharum officynarml.) is one of the most important sugar materials and bioenergy crops worldwide, and its production has strategic significance in guaranteeing sugar safety and sustainable development of energy. The production of cane is about 90% of the total sugar production in China. However, sugarcane production is threatened by a variety of diseases throughout the year, wherein smut (Sugarcane smut) caused by Saccharopolyspora spinosa (Sporisorium scitamineum) is a devastating systemic fungal disease that commonly occurs in various sugarcane areas worldwide, resulting in serious yield and sugar losses. The typical symptom of the disease is that pathogenic bacteria induce the abnormal differentiation of the meristem at the top of the cane stem to form a black whip structure (commonly called as 'black whip'). A large number of winter spores are generated in the black whip, and spread along with wind after maturation, so that the black whip becomes a main infection source for spreading diseases in the next year. Besides black whip, the infected plants also show abnormal phenotypes such as early germination, increased tillering, slender stalks, narrowed leaves and fading green leaves, and the normal growth and development of the sugarcane are seriously affected. The life history of the sorangium cellulosum is complex, and the sorangium cellulosum has polymorphism. The infection cycle starts from sexual coordination of haploid basidiomycetes of different mating types (Mat 1 and Mat 2) to form binuclear hyphae with infection capability. After the binuclear hyphae invade sugarcane buds, the binuclear hyphae grow and expand in host tissues, finally stimulate the hosts to form black whips and complete differentiation and maturation of winter spores in the black whips. Thus, sexual coordination of pathogenic bacteria is a key step in establishing infection, and formation of black whip and winter spores is a key step in completing and spreading disease circulation. At present, the prevention and treatment of sugarcane smut mainly depend on breeding and planting disease-resistant varieties. However, pathogenic bacteria are easy to mutate in physiological race, so that the original disease-resistant variety is often lost in resistance, for example, yue sugar 85-177 and Xintai sugar 10 which are disease-resistant varieties widely planted in China become disease-resistant varieties due to physiological race change (Shen Mokuan, 2004). Chemical control effects are limited and environmental residue risks exist, while biological control is still in a research and development stage and cannot be applied on a large scale. Therefore, the pathogenic mechanism of the Saccharopolyspora spinosa is deeply analyzed from the molecular level, and particularly, the key regulation and control path of sexual coordination and black whip development of the Saccharopolyspora spinosa has urgent need for developing novel, efficient and durable disease prevention and control strategies. In the process of researching the sexual coordination before and after gene expression mode of the sorangium cellulosum in the earlier stage, the team discovers that a specific gene in the sorangium cellulosum is named RWTD (Regulation ofWhip andTeliosporeDevelopment gene 1) and is specifically and highly expressed in binuclear hyphae formed after sexual coordination. The gene codes a protein predicted to contain four transmembrane domains, and presumably the gene can regulate the sexual coordination of the sorangium cellulosum, but no report on the functions of the protein and homologous proteins thereof exists before. Based on the above, the function of RWTD gene in the Saccharopolyspora spinosa is studied deeply, the functional characterization of four transmembrane proteins in the Saccharopolyspora spinosa can be filled, the understanding of sexual coordination, pathogenesis and disease symptom formation of the Saccharopolyspora spinosa is deepened, and a key novel regulation gene capable of comprehensively regulating and controlling the related proteins of the pathogenic bacteria cell membranes in the sexual coordination process is hopeful to be disclosed. Based on the gene, the research and development of a new method, a new medicament and a new product for preventing and controlling sugarcane smut by interfering the function of the gene provides a direct theoretical basis and a technical basis, and has great significance for solving the industrial problems of easy loss of resistant varieties, lack of prevention and control means and the like in the current prevention and control of sugarcane smut. Disclosure of Invention In order to solve the technical problems,