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Application of WRKY polypeptide Glyma02g39870 in promotion of salicylic acid biosynthesis and enhancement of disease resistance of plants

A technology of salicylic acid and plants, which is applied in the field of polypeptides that enhance plant disease resistance, and can solve the problems of rare plant salicylic acid

Active Publication Date: 2014-08-06
FUJIAN SANAN SINO SCI PHOTOBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few research reports on the regulation of plant salicylic acid synthesis.

Method used

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  • Application of WRKY polypeptide Glyma02g39870 in promotion of salicylic acid biosynthesis and enhancement of disease resistance of plants
  • Application of WRKY polypeptide Glyma02g39870 in promotion of salicylic acid biosynthesis and enhancement of disease resistance of plants
  • Application of WRKY polypeptide Glyma02g39870 in promotion of salicylic acid biosynthesis and enhancement of disease resistance of plants

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specific Embodiment approach

[0066] The present invention will be described in further detail below in combination with specific embodiments.

[0067] The methods used in the following examples are conventional methods unless otherwise specified. For specific steps, please refer to: "Molecular Cloning: A Laboratory Manual" (Sambrook J., Russell, David W., Molecular Cloning: A Laboratory Manual, 3rd edition, 2001, NY, Cold Spring Harbor). The primer synthesis and DNA sequence determination used were all carried out by Beijing Yingjun Biological Co., Ltd.

Embodiment 1

[0068] Example 1. Acquisition of Glyma02g39870 gene related to salicylic acid synthesis

[0069] The Phytozome database and literature were searched to obtain the sequence of the Glyma02g39870 gene (accession number: Glyma02g39870), and the forward primer was designed: 5'>GCTCTAGAATGGCATCTTCTTCTGGTAGT>3' and the reverse primer 5'>GCGAGCTCGGCATAGTAGAGACTCAAGGAA>3'. Extract the RNA of soybean leaves, obtain cDNA by reverse transcription, amplify the gene with TaKaRa’s PrimeSTAR HS high-fidelity enzyme method, construct it into the 326-FLAG expression vector, and transform the ligated product into Escherichia coli by heat shock method For the DH5α strain, select positive colonies and add them to 2.5ml of LB liquid medium containing 50mg / L ampicillin, and culture them in a shaker at 37°C and 180rpm for 12-16 hours. A small amount of plasmid was extracted, and after enzyme digestion and identification were correct, it was sent to Yingjun Biotechnology Co., Ltd. for sequence deter...

Embodiment 2

[0071] Embodiment 2, Arabidopsis salicylic acid synthetic key gene ICS1 gene promoter (Pro AtICS1 ) of the acquisition

[0072] Search the NCBI database and literature, obtain the sequence of AtICS1 (accession number: AT1G74710), analyze the promoter sequence according to the method of bioinformatics, and design forward primers according to the sequence analysis results: 5'>AACTGCAGTTATCCACGCTTTGTCACACA>3' and reverse Primer 5'>CGGGATCCCCATTGCAGAAATTCGTAAAGT>3'. The RNA of Arabidopsis leaves was extracted, cDNA was obtained by reverse transcription, the gene was amplified by TaKaRa’s PrimeSTAR HS high-fidelity enzyme method, and constructed into a 326-GFP expression vector, and the ligated product was transformed into Escherichia coli DH5α strain, select positive colonies and add them to 2.5ml LB liquid medium containing 100mg / L ampicillin, and cultivate them in a shaker at 37°C and 180rpm for 12-16 hours. A small amount of plasmid was extracted, and after enzyme diges...

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Abstract

The invention relates to a method for producing plants with enhanced disease resistance. The invention provides a WRKY polypeptide and a coding gene thereof. The WRKY polypeptide enhances the disease resistance of the plants through regulating the synthesis of salicylic acid in the plants.

Description

technical field [0001] The invention relates to a polypeptide for enhancing plant disease resistance, its coding nucleotide sequence and application. Some aspects of the present invention relate to introducing the nucleotide sequence encoding the WRKY polypeptide provided by the present invention into plant cells and expressing it, thereby promoting the synthesis of salicylic acid in plants and enhancing the disease resistance of plants. Background technique [0002] Plants often suffer from various diseases during the cultivation and production process. Some diseases pose a great threat to plant growth, such as root rot and gray spot. For crops, these diseases seriously impair their yield and quality. For plant diseases, at present, the main measures are to control them with drugs. Recently, people have begun to use abiotic inducers to induce plant disease resistance, and have been widely used in many important crops such as tobacco, potato, tomato, cucumber, bean, and ri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N5/10C07K14/415A01H5/00
Inventor 金京波蔡斌周晓锋
Owner FUJIAN SANAN SINO SCI PHOTOBIOTECH CO LTD