CN-121555474-B - Phytophthora GLS1 protein, and encoding gene and application thereof

Abstract

The invention discloses a glucosidase protein (GLS 1) from phytophthora plant pathogenic oomycetes, and a coding gene and application thereof. The PsGLS protein provided by the invention is a protein with a sequence shown as SEQ ID No.1 in phytophthora sojae (Phytophthora sojae), and the coding gene is shown as SEQ ID No. 2. Experiments prove that the protein provided by the invention plays an important role in the growth and development process of the phytophthora sojae, the hypha growth rate, sporangium number, zoospore yield and resting spore germination rate of the phytophthora sojae after PsGLS gene knockout are obviously reduced, and pathogenic bacteria pathogenicity is obviously influenced. The glucosidase inhibitor castanospermine has an inhibitory effect on Phytophthora sojae silk growth. In conclusion, the PsGLS gene of the invention has high application value in controlling epidemic diseases caused by phytophthora, and provides a molecular target for the research and development of novel bactericides in the future.

Inventors

• ZHANG CAN
• LIU ZHANYUN
• GENG SHAN
• Chang Zhengjie
• Yang Lilan
• LIU XILI
• LI XINPING
• ZHANG FAN

Assignees

• 中国农业大学

Dates

Publication Date

20260505

Application Date

20260121

Claims (4)

1. 1. The application of the biological material related to the GLS1 protein coding gene shown in SEQ ID No.1 is characterized in that the application is any one or more of the following 1) -3): 1) Use in inhibiting the production of sporocysts of phytophthora sojae; 2) Application in inhibiting zoospore yield of Phytophthora sojae; 3) The application of the composition in reducing pathogenicity of phytophthora sojae; Wherein the biological material related to the coding gene is any one of the following D1-D2): D1 A nucleic acid molecule that inhibits expression of a gene encoding the GLS1 protein; d2 An expression cassette and a recombinant vector comprising the nucleic acid molecule of D1).
2. 2. The use according to claim 1, wherein the coding gene is a DNA molecule represented by the nucleotide sequence shown in SEQ ID No.2 of the sequence listing.
3. 3. The use according to claim 1 or 2, comprising effecting said use of 1) -3) by knocking out sequences in said coding gene.
4. 4. A method for reducing the activity of phytophthora sojae comprises the steps of knocking out the coding gene shown in SEQ ID No.2 or inhibiting the activity of GLS1 protein shown in SEQ ID No.1 in phytophthora sojae; Wherein, the activity of reducing the phytophthora sojae is to reduce the sporangium yield and/or zoospore yield of phytophthora sojae and/or pathogenicity to a host.

Description

Phytophthora GLS1 protein, and encoding gene and application thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to GLS1 protein from phytophthora sojae (Phytophthora sojae), and a coding gene and application thereof. Background Oomycetes (Oomycete) are widely distributed, and the host range thereof covers plant kingdoms and animal kingdoms, wherein the plant pathogenic oomycetes account for about 60% of the total oomycetes and can be classified into Phytophthora (Phytophthora), pythum (Pythum), peronospora (Peronospora) and the like. The infection of crops by phytopathogenic oomycetes is of a wide variety and causes great economic losses to agricultural production, wherein phytopathogenic oomycetes pathogen (Phytophthora spp.) is particularly serious in agricultural production. Under natural conditions, the life history of phytophthora is divided into asexual and sexual stages. In the sexual reproduction stage, the oospores are formed through the cooperation of the same mass, the environment is resisted, the oospores survive in the soil for a plurality of years, and when the environment is proper, the oospores germinate and infect hosts to become primary infection sources. In the asexual propagation stage, phytophthora sporocysts generate and release zoospores in a room temperature and liquid environment, rapidly differentiate into resting spores after a short period of rest, and further germinate and infect hosts. As phytopathogenic oomycetes causing typical soil-borne diseases, phytophthora sojae (Phytophthora sojae) can cause the occurrence of soybean stem and root rot (PRR), which can be detrimental during the whole growth period of soybeans. The infected soybean plants show typical symptoms of decay from the root, gradually spreading from bottom to top along the stems, forming macroscopic brown lesions in the stems, especially at the base of the stems, with serious economic losses in soybean yield. The phytophthora sojae has a plurality of physiological seeds, has rapid variation and strong pathogenicity, severely limits the development and the utilization of phytophthora resistance genes, has long effective time for biological control and has higher requirements on manpower and material resources. Chemical control is still one of the most effective methods for controlling oomycete diseases at present. With the deep analysis of the action mechanism of the bactericide, targeting the key binding site has become a core strategy for creating novel oomycete inhibitors, so that the development of the oomycete bactericide based on novel targets is particularly important and urgent. In the process of infecting a host by plant pathogenic bacteria, defensive small molecules generated by plants can interfere with the normal infecting process of the pathogenic bacteria, and the pathogenic bacteria resist the reverse stress in the infecting process and are important for the normal infecting of the host. The glucosidase protein (GLS 1) is used as a key protein for quality control of endoplasmic reticulum, and can promote correct folding of the protein by transferring glucose molecules on peptide chains, but the functions of the protein in oomycetes such as phytophthora have not been reported yet. In summary, zoospores of phytophthora and the capability of infecting hosts are important factors influencing the occurrence and development of diseases, and if the process of phytophthora can be blocked by influencing the key protein of phytophthora, the damage caused by phytophthora can be controlled. Disclosure of Invention The invention identifies 1 glucosidase GLS1 (mannosyl-oligosaccharide glucosidase) in phytophthora sojae, the protein belongs to glycoside hydrolase 63 family (GH 63 family), and contains 1 glycosyl hydrolase 63 family N-terminal domain (Glyco _hydro_63N super family) and one trehalose family domain (TREHALASE SUPER FAMILY), and is named PsGLS1, Through the research of the inventor, the GLS1 in the phytophthora sojae is closely related to the growth rate of phytophthora sojae silk, the sporangium quantity, the zoospore yield and the resting spore germination rate, and simultaneously the response of pathogenic bacteria to endoplasmic reticulum stress is influenced. Therefore, the ability of phytophthora sojae to infect a host can be weakened by regulating and controlling the function of GLS1 protein, so that the occurrence and development of phytophthora sojae root rot can be controlled. The sequence of the GLS1 protein (PsGLS 1) in the phytophthora sojae is shown as SEQ ID No.1 in a sequence table, the SEQ ID No.1 (PsGLS) in the sequence table consists of 905 amino acid residues, the DNA sequence (coding gene and cDNA) required for coding the PsGLS protein can be specifically shown as SEQ ID No.2 in the sequence table, the SEQ ID No.2 (PsGLS 1) in the sequence table consists of 2718 nucleotides, and the protein PsGLS1 shown as the SEQ ID No.1 in t