CN-122011141-A - Huang-Huai sacculus mould GIP12 effector protein, coding gene and application thereof

Abstract

The invention belongs to the technical field of genetic engineering, and discloses a sacculus flavus GIP12 protein, a coding gene and application thereof, wherein the amino acid sequence of Huang Huaiqiu sacculus flavus GIP12 effector protein is shown as SEQ ID NO.2, and the nucleotide sequence of nucleic acid or gene for coding the effector protein is shown as SEQ ID NO. 1. The cloning of the G.chrysotium GIP12 protein gene and the transient expression of tobacco prove that the G.chrysotium GIP12 protein plays an important role in the process of improving plant disease resistance.

Inventors

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

Assignees

• 江苏农林职业技术学院

Dates

Publication Date

20260512

Application Date

20260205

Claims (10)

1. 1. The Huang-Huai sacculus fungus GIP12 effector protein is characterized in that the amino acid sequence of the Huang Huaiqiu sacculus fungus GIP12 effector protein is shown as SEQ ID NO. 2.
2. 2. A nucleic acid or gene encoding the Huang Huaiqiu cyst GIP12 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 sporadic GIP12 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 GIP12 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 GIP12 effector protein as defined in claim 1, nucleic acid or gene as defined in claim 2, expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacteria as defined in claim 3 for the preparation of a reagent for inducing plant allergy.
9. 9. Use of Huang Huaiqiu sporangium GIP12 effector protein as defined in claim 1, a nucleic acid or gene as defined in claim 2, an expression cassette, a recombinant expression vector, a transgenic cell line or a transgenic recombinant bacterium as defined in 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 mould GIP12 effector protein, coding gene and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a Huai sacculus mould GIP12 effector protein, a coding gene and application thereof. Background Plants are subjected to attack by various microbial pathogens throughout the life cycle, during the co-evolution of plants and pathogens, adaptive pathogens evolve various effector proteins that attack the plants, whereas non-toxic proteins (Avr effectors) are a class of proteins that can be recognized directly or indirectly by plant R proteins, and can trigger allergic reactions (HYPERSENSITIVE RESPONSE, HR). The purpose of the HR response is to limit further spread and infection of pathogens. Plants form a physical barrier through cell death, prevent the transmission of pathogens, secrete antibacterial substances, active oxygen and the like to inhibit the growth of pathogens, and are also important signals for stimulating the immunity of plants. Glucose-dependent insulinotropic polypeptide effector proteins (GIPs) are a family of secreted protein genes specific to oomycetes (e.g., phytophthora, downy mildew), encoding "regulatory-interfering" bifunctional proteins comprising specific functional domains and signal regulatory modules. In the infection process, GIP regulates host sugar metabolism and immune balance by targeting a key signal path in a host cell to build a proper infection microenvironment for a pathogen, and serves as an effector to be recognized by a plant immune system to trigger complex immune response (hormone signal reprogramming, defense gene expression regulation and cell metabolism remodeling). Because of the core regulation and control effects of the GIP in oomycete pathogenic strategy and host immune escape, GIP has become a key target for analyzing oomycete-plant interaction molecular mechanism and is explored as a potential target protein of novel crop disease prevention and control strategy, and is not reported in sacculus mildew at present. Huang Huaiqiu cyst mould (Globisporangium huanghuaiense) is a pathogenic oomycete and can cause root rot of plants such as wheat, rice, soybean and the like, so that serious economic loss is caused. Disclosure of Invention Object of the invention A first object of the present invention is to provide Huang Huaiqiu cyst GIP 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 GIP 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 present invention is to provide the use of the above Huang Huaiqiu cyst GIP 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 saccorium chrysosporium GIP12 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 GIP12 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 GIP12 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 GIP12 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 GIP12 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 GIP12 effector protein, nucleic acid or gene, expression cassette, recombinant expression vector, transgenic cell line or transgenic recombinant bacteria of the invention are used for 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