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Application of elicitor protein derived from the fungus Cladobrachia in the control of cassava bacterial wilt

A technology of elicitor and Cladosporium, applied in the field of molecular biology, can solve problems such as undiscovered coding genes

Active Publication Date: 2021-08-31
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, many new protein elicitors have been discovered, but there is no report on the elicitor protein and its encoding gene that can induce plant disease resistance in the fungus of the genus Cladostratum

Method used

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  • Application of elicitor protein derived from the fungus Cladobrachia in the control of cassava bacterial wilt
  • Application of elicitor protein derived from the fungus Cladobrachia in the control of cassava bacterial wilt
  • Application of elicitor protein derived from the fungus Cladobrachia in the control of cassava bacterial wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the cloning of the coding gene of elicitor protein SbES and its cDNA

[0043]A high-efficiency biocontrol strain isolated from Brachiaria leaves in our laboratory was identified as Sarocladium brachiariae and named as Sarocladium brachiariae HND5 (Liu X, Guo Z, Huang G: Sarocladium brachiariae sp. nov., an endophytic fungus isolated from Brachiaria brizantha. Mycosphere 2017, 8:827-834.). The inventors of the present invention, on the basis of the whole genome sequencing data of the strain, compared and analyzed the elicitor sequences reported by relative fungi, found and identified the potential for improving plant resistance and inducing plant defense response activity. Elicitor protein gene.

[0044] The method for obtaining the full-length coding DNA of the elicitor protein gene of the present invention and the full-length cDNA fragment is as follows:

[0045] Using the total DNA extracted from the Sarocladium brachiariae HND5 strain as a template, d...

Embodiment 2

[0052] Embodiment 2, eukaryotic expression and purification of SbES protein

[0053] 1. Optimization of cDNA sequence based on codon preference of Pichia pastoris

[0054] In order to use Pichia pastoris X-33 strain for heterologous expression of SbES protein, according to the codon preference of Pichia pastoris, the signal peptide and propeptide encoding genes of SbES protein were excluded, and the 316-1164 nucleotides of sequence 2 were Codon optimization was carried out with acid to obtain sequence 4, which is encoded from the protein shown in amino acid 106-387 of the amino terminal of sequence 1 in the sequence listing. Sequence 4 was synthesized by Beijing Huada Biotechnology Co., Ltd.

[0055] 2. Construction of eukaryotic expression vector

[0056] The primers SbES-PF and SbES-PR were paired, and the sequence 4 was used as a template, and the amplified fragment was connected to the T vector, named pMD18-T / SbESp; pMD18-T / SbESp was double-digested with EcoRI and KpnⅠ, ...

Embodiment 3

[0066] Example 3, Evaluation of the control effect of SbES protein on cassava bacterial wilt

[0067] Pathogen strain: Xanthomonas axonopodis strain XamGX11 (Construction and quality evaluation of the transposon library of cassava bacterial wilt [J]. Journal of Tropical Crops, 2013, 34(6): 1144-1148.) , the public can obtain the strain from the Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences.

[0068] Cassava variety: SC10

[0069]Medicament: recombinant protein SbES (0.1 mg / mL), Artelin (3% oligosaccharide protein·3% ​​Alternaria activator protein WP) (Institute of Plant Protection, Chinese Academy of Agricultural Sciences), and PBS Buffer as the control.

[0070] Chemical treatment: Adjust the concentration of the recombinant protein His-SbES to 0.1 mg / mL, spray 10 potted cassava seedlings for 3 times with an interval of 3 days, and treat with Alterin: Alterin (3% oligosaccharide protein·3% ​​chain spacer Sporogenin WP) was ...

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Abstract

The invention discloses the application of an elicitor protein derived from a fungus of the genus Brachidella to prevent and treat cassava bacterial wilt disease. The application is an application in inducing cassava to produce disease resistance to bacterial wilt disease or improving cassava disease resistance with the elicitor protein derived from the fungus of the genus Brachiaria. The elicitor protein is a protein with one of the following amino acid residue sequences: 1) the amino acid residue sequence of SEQ ID No. 1 in the sequence listing; 2) the amino acid residue sequence of SEQ ID No. 1 in the sequence listing A protein derived from SEQ ID No. 1 that has undergone substitution and / or deletion and / or addition of one or several amino acid residues and has an elicitor protein function. Experiments have proved that the protein elicitor SbES can induce cassava production or improve plant resistance to target pathogens, reduce disease incidence and disease index, provide a new way to improve plant disease resistance, and have broad application prospects in agricultural production .

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to the application of an elicitor protein derived from a fungus of the genus Brachia and its coding gene in preventing and treating cassava bacterial wilt. Background technique [0002] Protein elicitors are derived from some proteins secreted by bacteria and fungi, mainly including Harpin, Cryptogein, and Activator. By inducing the expression of plant defense-related genes and activating defense signaling pathways in plants, such as salicylic acid, etc., plants can be stimulated to acquire resistance, and plant self-immunity can be enhanced to resist the infection of pathogenic microorganisms. Many new protein elicitors have been discovered so far, but there is no report on the elicitor protein and its encoding gene that can induce plant disease resistance in the fungus of the genus Cladopa. Contents of the invention [0003] The high-efficiency biocontrol strain isolated from the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/37
CPCA01N63/50A01N63/60C07K14/37C12N15/815
Inventor 李超萍杨扬时涛王国芬蔡吉苗李博勋黄贵修
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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