CN-122011144-A - Huang-Huai sacculus mildew EPI effector protein, coding gene and application thereof

Abstract

The invention belongs to the technical field of genetic engineering, and discloses a coding gene and application of a balloon-mould flavescens EPI effector protein, wherein the Huang Huaiqiu balloon-mould EPI effector protein is EPI1, the amino acid sequence of the EPI effector protein is shown as SEQ ID NO.2, and the nucleotide sequence of a nucleic acid or gene for coding the EPI effector protein is shown as SEQ ID NO. 1. The cloning of the sacculus flavescens EPI1 protein gene and the transient expression of tobacco prove that the sacculus flavescens EPI1 protein plays an important role in the process of improving plant disease resistance.

Inventors

• PENG JINFENG
• LV WANQING
• CHEN JIAJIA
• YU JIAN
• SUN YAYA
• FU DANYANG
• ZHENG XIANGRONG
• WU XIANGQIAN
• LI SONG
• HUANG XIAOXIAO

Assignees

• 江苏农林职业技术学院

Dates

Publication Date

20260512

Application Date

20260205

Claims (10)

1. 1. The balloon mold EPI effector protein for Huang Huai is characterized in that the Huang Huaiqiu balloon mold EPI effector protein is EPI1, and the amino acid sequence of the EPI effector protein is shown as SEQ ID NO. 2.
2. 2. A nucleic acid or gene encoding the Huang Huaiqiu cyst mould EPI effector protein of claim 1, wherein the nucleotide sequence of said nucleic acid or gene is shown in SEQ ID No. 1.
3. 3. An expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacterium comprising a nucleic acid or gene of the Huang Huaiqiu sporangium EPI effector protein of claim 2.
4. 4. The method for constructing a recombinant expression vector according to claim 3, wherein the expression vector is obtained by introducing a gene encoding Huang Huaiqiu sporangium EPI effector protein into a plant expression vector pCAMBIA 1300-FLAG.
5. 5. The construction method according to claim 4, wherein the nucleotide sequence of the primer used in the construction process of the expression vector is shown in SEQ ID NO. 3-4.
6. 6. The recombinant bacterium according to claim 3, wherein the recombinant expression vector according to claim 3 is inserted into E.coli and the recombinant bacterium is obtained by screening.
7. 7. The transgenic recombinant strain of claim 6, wherein the escherichia coli is a DH5 a competent cell.
8. 8. Use of Huang Huaiqiu sporangium EPI effector protein according to claim 1, a nucleic acid or gene according to claim 2, an expression cassette, a recombinant expression vector, a transgenic cell line or a transgenic recombinant bacterium according to claim 3 for the preparation of a reagent for inducing plant allergy.
9. 9. Use of Huang Huaiqiu sporangium EPI effector protein according to claim 1, a nucleic acid or gene according to claim 2, an expression cassette, a recombinant expression vector, a transgenic cell line or a transgenic recombinant bacterium according to claim 3 for the preparation of a reagent for increasing plant resistance.
10. 10. The use according to claim 9, wherein said increasing plant resistance is increasing the resistance of a plant to phytophthora capsici infection.

Description

Huang-Huai sacculus mildew EPI effector protein, coding gene and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a sacculus flavus EPI effector protein, a coding gene and application thereof. Background Huang Huaiqiu the cyst mould (Globisporangium huanghuaiense) belongs to oomycetes, has strong pathogenicity and quick transmission, mainly infects plant root systems, damages the structure and the function of the plant, causes the plant to have symptoms of wilting, yellowing, growth stagnation and the like, and causes the plant to die when serious. The existing control means still mainly depend on chemical pesticides, but the problems of enhanced drug resistance of pathogenic bacteria, deterioration of soil ecological environment, exceeding of agricultural product residues and the like are easily caused after long-term use, so that the urgent requirements of green and high-quality development of agriculture are difficult to meet. Therefore, the exploration of new strategies for green prevention and control based on crop autoimmune mechanisms has become a key direction of industrial sustainable development. Studies have shown that Huang Huaiqiu sporangium secretes a range of effector proteins into the host during infection, which are key weapons for its pathogenesis and also play a central role in pathogen interactions with plants. Among them, the elicitor proteins that are recognized by the plant immune system and activate the defensive response have great potential to be novel biogenic immune elicitors. Although elicitors as an important class of plant immunomodulators have shown good promise in improving crop disease resistance, current excavation of saccharum chrysosporium-specific elicitors is still very limited. Therefore, the systematic identification of key elicitor proteins of Huang Huaiqiu sporangium can provide key theoretical basis and core technical support for developing novel immune elicitors with independent intellectual property rights, creating disease-resistant germplasm resources and establishing a green accurate prevention and control technical system. Disclosure of Invention Object of the invention A first object of the present invention is to provide a Huang Huaiqiu cyst EPI effector protein that can induce plant allergic reactions (HYPERSENSITIVE RESPONSE, HR). A second object of the present invention is to provide a nucleic acid or gene encoding the above Huang Huaiqiu cyst membrane EPI effector protein. The third object of the present invention is to provide an expression cassette, a recombinant expression vector, a transgenic cell line or a transgenic recombinant bacterium containing the above nucleic acid or gene, and construction methods of the recombinant expression vector and the transgenic recombinant bacterium. The fourth object of the invention is to provide the use of the Huang Huaiqiu sporangium EPI effector protein, nucleic acid or gene, expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacterium. In order to achieve the above purpose, the present invention adopts the following technical scheme: the invention discloses a yellow-Huai sacculus mildew EPI effector protein, wherein Huang Huaiqiu sacculus mildew EPI effector protein is EPI1, and the amino acid sequence of the EPI effector protein is shown as SEQ ID NO. 2. The nucleotide sequence of the nucleic acid or the gene for encoding the Huang Huaiqiu cyst membrane EPI effector protein is shown as SEQ ID NO. 1. The invention relates to an expression cassette, a recombinant expression vector, a transgenic cell line or a transgenic recombinant bacterium containing the nucleic acid or the gene of the Huang Huaiqiu cyst membrane EPI effector protein. The invention relates to a construction method of a recombinant expression vector, which is obtained by introducing a gene for encoding Huang Huaiqiu cyst membrane EPI effector protein into a plant expression vector pCAMBIA 1300-FLAG. The nucleotide sequence of the primer used in the construction process of the expression vector is shown in SEQ ID NO. 3-4. The transgenic recombinant strain is obtained by inserting the recombinant expression vector into escherichia coli and screening. Wherein the E.coli is DH5 alpha competent cells. The Huang Huaiqiu cyst membrane EPI effector protein, nucleic acid or gene, expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacteria are applied to the preparation of plant allergic reaction inducing reagent. The Huang Huaiqiu cyst membrane EPI effector protein, nucleic acid or gene, expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacteria are applied to preparation of plant resistance improving agent. Wherein, the improvement of plant resistance is the improvement of the resistance of plants to phytophthora capsici infe