CN-122012551-A - Application of disease-resistant related protease GmRD21A gene and encoding protein thereof in plant epidemic disease resistance

Abstract

The invention belongs to the technical field of plant molecular biology, and discloses a soybean disease-resistant related protease GmRD21A gene and a protein encoded by the same, and application of the soybean disease-resistant related protease GmRD21A gene in plant epidemic disease resistance. The nucleotide sequence of the protease GmRD21A gene is shown as SEQ ID NO.1, and the amino acid sequence of the polypeptide encoded by the protease GmRD21A gene is shown as SEQ ID NO. 2. According to the invention, the protein mass spectrum analysis of the phytophthora sojae pectin lyase PsPL is carried out, so that the degradation of the pectin lyase secreted by phytophthora sojae by the protease GmRD21A can resist the attack of pathogenic bacteria pectin lyase on the plant cell wall, and meanwhile, the over-expression of the GmRD21A gene in soybean can obviously improve the resistance of plants to phytophthora sojae. The invention can be applied to the improvement of crop breeding epidemic disease resistance, and is hopeful to improve the disease resistance of plants to epidemic disease, thereby achieving the purposes of increasing yield and reducing medicine.

Inventors

• SUN GUANGZHENG
• WANG YUANCHAO
• ZHU QINSHENG

Assignees

• 南京农业大学

Dates

Publication Date

20260512

Application Date

20260311

Claims (10)

1. 1. The disease-resistant related protease GmRD21A gene is characterized in that the nucleotide sequence of the disease-resistant related protease GmRD21A gene is shown in any one of the following, 1) Nucleotide sequence shown as SEQ ID NO.1 and its complementary sequence, or 2) The nucleotide sequence shown as SEQ ID NO.1 is substituted, deleted and/or added with one or more nucleotides to express the nucleotide sequence of the same functional protein, or 3) Nucleotide sequence which hybridizes under stringent conditions with the sequence shown in SEQ ID No.1, or 4) A sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity with the nucleotide sequence shown in SEQ ID No.1, and a polypeptide encoded thereby having a function of improving plant disease resistance.
2. 2. A polypeptide coded by disease-resistant related protease GmRD21A gene is characterized in that the amino acid sequence of the polypeptide is shown in any one of the following, 1) An amino acid sequence as shown in SEQ ID NO.2, or 2) An amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO.2, said polypeptide having a function of improving disease resistance of a plant, or 3) An amino acid sequence obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence shown in SEQ ID NO.2, and the amino acid sequence has the function of regulating plant disease resistance.
3. 3. A recombinant expression vector comprising the disease-resistant related protease GmRD21A gene of claim 1.
4. 4. A host cell comprising the disease-resistant related protease GmRD21A gene of claim 1, or comprising the recombinant vector of claim 3.
5. 5. The use of the disease-resistant related protease GmRD21A gene of claim 1, the polypeptide of claim 2, the recombinant expression vector of claim 3, or the host cell of claim 4, wherein the use is any of, 1) Improving the degradation capability of the plant to pectin lyase secreted by pathogenic bacteria; 2) Improving the resistance of plants to phytophthora; 3) Breeding plant variety with epidemic disease resistance.
6. 6. The use according to claim 5, wherein said disease-resistant or said pathogenic bacteria include, but are not limited to, phytophthora sojae, phytophthora nicotianae, phytophthora infestans and phytophthora capsici.
7. 7. Use according to claim 5, wherein the plant is a leguminous plant or a solanaceous plant, preferably soybean, tomato, potato or tobacco.
8. 8. A method for improving the basic disease resistance of plants, which comprises transferring the disease resistance related protease GmRD21A gene, the polypeptide, the recombinant expression vector or the host cell into the plant tissue.
9. 9. The method according to claim 8, wherein the plant is a leguminous plant or a solanaceous plant, preferably soybean, tomato, potato or tobacco.
10. 10. Method for breeding plant disease-resistant variety and detecting disease-resistant related protease in plant variety to be detected GmRD21A The expression level of the gene, if the disease-resistant related protease GmRD21A The detection primer sequences adopted in detection are preferably shown as SEQ ID NO.3 and SEQ ID NO.4, wherein the upstream primer is shown as SEQ ID NO.3, and the downstream primer is shown as SEQ ID NO. 4.

Description

Application of disease-resistant related protease GmRD21A gene and encoding protein thereof in plant epidemic disease resistance Technical Field The invention belongs to the technical field of plant molecular biology, and particularly discloses an application of a disease-resistant related protease GmRD21A gene and a protein encoded by the same in plant epidemic disease resistance. Background Phytophthora seriously harms crops such as soybeans, potatoes and the like, and causes huge economic loss. Phytophthora belongs to the phylum oomycetes, is known to be the 'root rot' of important diseases on various crops, for example, the root rot caused by the phytophthora sojae (Phytophthora sojae) is a destructive disease on soybean production, and potato late blight bacteria cause the explosion of Irish hunger in the 19 th century, and the damage to crops such as potatoes is huge every year. The economic loss of phytophthora caused to crops in China every year is up to 160 hundred million yuan, and the production of the crops in China is seriously influenced. At present, the pathogenic bacteria are easy to generate drug resistance and the crop resistance is easy to lose in production for preventing and controlling epidemic diseases. Therefore, research on plant disease resistance mechanism is developed, and the method has important significance for improving crop resistance and designing plant epidemic disease prevention and control strategies. The plant apoplast is the main battlefield of plants against pathogens, where cysteine proteases secreted by plants play a key role in plant disease resistance. For example, papain-like cysteine protease (Papain-LIKE CYSTEINE proteases PLCPs) in corn can cleave Prozip precursor and release of the Zip1 small peptide can activate the salicylic acid signaling pathway. Therefore, the plant protease is utilized and modified, is a new way for durable and safe prevention and control of plant diseases, and meets the requirements of green development and sustainable development of the current agriculture. Disclosure of Invention The invention mainly aims to provide an application of disease-resistant related protease GmRD21A gene capable of degrading pathogenic bacteria pectin lyase and a coded protein thereof in plant disease resistance. In order to achieve the above object, the present invention provides the following technical solutions. In one aspect, the invention provides a disease-resistant related protease GmRD21A gene, the nucleotide sequence of the disease-resistant related protease GmRD21A gene is shown in any one of the following, 1) Nucleotide sequence shown as SEQ ID NO.1 and its complementary sequence, or 2) The nucleotide sequence shown as SEQ ID NO.1 is substituted, deleted and/or added with one or more nucleotides to express the nucleotide sequence of the same functional protein, or 3) Nucleotide sequence which hybridizes under stringent conditions with the sequence shown in SEQ ID No.1, or 4) A sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity with the nucleotide sequence shown in SEQ ID No.1, and a polypeptide encoded thereby having a function of improving plant disease resistance. On the other hand, the invention also provides a polypeptide coded by the disease-resistant related protease GmRD21A gene, the amino acid sequence of the polypeptide is shown in any one of the following, 1) An amino acid sequence as shown in SEQ ID NO.2, or 2) An amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO.2, said polypeptide having a function of improving disease resistance of a plant, or 3) An amino acid sequence obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence shown in SEQ ID NO.2, and the amino acid sequence has the function of regulating plant disease resistance. On the other hand, the invention also provides a recombinant expression vector, which comprises the disease-resistant related protease GmRD21A gene. In another aspect, the invention also provides a host cell comprising the disease-resistant related protease GmRD21A gene, or comprising the recombinant vector. In another aspect, the invention also provides the use of the disease-resistant related protease GmRD21A gene, or the polypeptide, or the recombinant expression vector, or the host cell, the use being any of, 1) Improving the basic disease resistance of plants, 2) improving the degradation capability of the plants to pectin lyase secreted by pathogenic bacteria, and 3) breeding disease-resistant plant varieties. Further, the disease-resistant or pathogenic bacteria include, but are not limited to, phytophthora sojae, Further, the pathogenic bacteria resistant to or susceptible to disease include, but are not limited to, phytophthora sojae, phytophthora nicotianae, phytophthora infestans and phytophthora capsici. Further, the pla