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Wound-induced gene of soybeans, and coded protein and application thereof

A protein-encoding and gene-encoding technology, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as agricultural production losses, and achieve the effect of increasing plant resistance to bacterial diseases

Active Publication Date: 2018-11-16
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pseudomonas syringae is a large group of extremely popular plant pathogenic bacteria, which can infect more than 300 economic crops such as leguminous, cruciferous, and solanaceous, and the incidence of plant diseases caused by it ranks among the highest among bacterial plant diseases First, causing huge losses to agricultural production

Method used

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  • Wound-induced gene of soybeans, and coded protein and application thereof
  • Wound-induced gene of soybeans, and coded protein and application thereof
  • Wound-induced gene of soybeans, and coded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Cloning GmWIP Gene

[0023] Soybean ( Glycine max ) a gene encoding a wound-inducible protein was highly expressed in the nodule transcriptome GmWIP . Using the soybean nodule cDNA as a template, the polymerase chain reaction (PCR) method was used to clone GmWIP CDS sequence of the gene. The full length of the CDS sequence is 255bp, encoding a protein consisting of 84 amino acid residues. This result correlates with that published by the Soybean Genome Database (https: / / phytozome.jgi.doe.gov / pz / ) GmWIP The relevant sequence information of the genes is completely consistent.

Embodiment 2

[0024] Example 2: Construction GmWIP constitutive expression vector

[0025] First PCR clone with upstream and downstream primers with restriction enzyme sites Xho I and BamH I respectively GmWIP CDS sequence, through a series of enzyme digestion-ligation reactions GmWIP The CDS was ligated into the intermediate vector pA7 to obtain 35S:GmWIP - pA7 intermediate vector. Then use Hind III and EcoR I double digestion reaction to 35S:GmWIP The fragment was excised and connected to the binary vector pCAMBIA2301 to obtain the final 35S:GmWIP - pCAMBIA2301 vector.

Embodiment 3

[0026] Embodiment three: GmWIP Transformation of Arabidopsis thaliana with constitutive expression vector

[0027] Complete the above construction 35S:GmWIP -The pCAMBIA2301 vector was transformed into Agrobacterium strain GV3101 by heat shock method, and then the target gene was transformed into Arabidopsis thaliana through flower dipping method through Agrobacterium infection. The transgenic positive plants were obtained by kanamycin plate screening, and then T3 generation homozygous transgenic plants were obtained after two generations of self-propagation.

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Abstract

The invention relates to a wound-induced gene of soybeans, and a coded protein and application thereof. The gene is a nucleotide sequence as shown in SEQ ID No. 1, and the protein coded by the gene isan amino acid sequence as shown in SEQ ID No. 2. According to the invention, the brand novel wound-induced gene GmWIP is cloned in soybeans for the first time; and constitutive expression of the geneGmWIP in plants proves that the gene and the protein coded by the gene have the function of improving the disease resistance of plants.

Description

technical field [0001] The invention relates to a soybean injury-inducing gene, its encoded protein and its application. Background technique [0002] Plant bacterial diseases are diseases caused by infection by bacterial pathogens, such as soft rot, canker, bacterial wilt, etc. Bacterial diseases are one of the most serious diseases that endanger agricultural production. When an epidemic breaks out, it often leads to a large-scale reduction in crop production, or even failure. At present, the prevention and control measures for bacterial diseases mainly rely on the use of pesticides. However, on the one hand, large-scale use of pesticides will easily cause the plants to develop drug resistance and lead to poor control effects; on the other hand, the residues of pesticides, especially antibiotic pesticides, have become a threat. Huge hidden danger to human and animal health. In recent decades, a large number of studies have shown that there is a natural immune system in pl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/20
CPCC12N15/8281C07K14/415
Inventor 于亮亮罗利闫军辉王亚雯
Owner SHANGHAI UNIV
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