CN-122011142-A - Huang-Huai sacculus mildew EPI7 protein, coding gene and application thereof

Abstract

The invention belongs to the technical field of genetic engineering, and discloses a sacculus flavus EPI7 protein, a coding gene and application thereof, wherein the amino acid sequence of the Huang Huaiqiu sacculus flavus EPI7 protein is shown as SEQ ID NO.2, and the nucleotide sequence of a nucleic acid or a gene for coding the EPI7 protein is shown as SEQ ID NO. 1. The cloning of the sacculus flavescens EPI7 protein gene and the transient expression of tobacco prove that the sacculus flavescens EPI7 protein plays an important role in the process of improving plant disease resistance.

Inventors

• CHEN JIAJIA
• PENG JINFENG
• YU JIAN
• SUN YAYA
• FU DANYANG
• HUANG XIAOXIAO
• ZHENG XIANGRONG
• LI SONG
• QIU XIUWEN

Assignees

• 江苏农林职业技术学院

Dates

Publication Date

20260512

Application Date

20260205

Claims (10)

1. 1. The sacculus flavomorphus EPI7 protein is characterized in that the amino acid sequence of the Huang Huaiqiu sacculus flavomorphus EPI7 protein is shown as SEQ ID No. 2.
2. 2. A nucleic acid or gene encoding the Huang Huaiqiu cyst mould EPI7 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 sporadic mold EPI7 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 EPI7 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 EPI7 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 EPI7 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 EPI7 protein, coding gene and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a balloon mold EPI7 protein of Huang Huai, a coding gene and application thereof. Background Effector proteins are a class of virulence molecules secreted by pathogenic bacteria during infection of host plants that disrupt the immune system of plant cells, thereby promoting infection by pathogenic bacteria. Huang Huaiqiu the cyst mould (Globisporangium huanghuaiense) is used as a plant pathogenic oomycete, and can secrete a plurality of effector proteins in the infection process of a host, wherein the effector proteins can interact with different targets in the host to interfere the immune response of plants or change the metabolic pathway of the plants, so that the infection and the colonization of the pathogenic bacteria are promoted. However, once these effector proteins are recognized by the immune receptor of the host, they activate the defensive response of the plant, limiting the spread of pathogenic bacteria. Thus, the effector protein plays a dual role in the interaction of pathogenic bacteria and hosts, and not only affects the pathogenicity of the pathogenic bacteria, but also regulates and controls the expression of plant disease resistance. The function and action mechanism of effector protein are studied deeply, which is not only helpful for revealing the pathogenic mechanism of pathogenic bacteria and the immune response mechanism of plants, but also provides important theoretical basis for designing a durable and efficient disease prevention and control strategy based on host-pathogen interaction relation. EPI effector proteins are secreted functional proteins with conserved Kazal type structural domains in oomycetes, promote pathogen infection by targeted inhibition of host disease course related protease activity and interference of plant immune signaling, but part of EPI proteins can be used as excitons to be recognized by plant specific receptors, rapidly trigger allergic necrosis reaction (HR) and activate systemic immune response, and have the potential of being developed into plant immune elicitors. 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 amino acid sequence of the Huang Huaiqiu sporangium EPI7 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 EPI7 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 sporangium EPI7 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 EPI7 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 EPI7 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 sporangium EPI7 protein, nucleic acid or gene, expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacteria are used in preparing plant resistance reagent. Wherein, the improvement of plant resistance is the improvement of the resistance of plants to phytophthora capsici infection. Compared with the prior art, the invention has the remarkable advantages that 1 gene EPI7 which is derived from the sacculus flavdi