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Plant disease-resistant protein GhWRKY40, coding gene and application thereof

A technology for encoding and planting, applied in the field of plant disease resistance protein GhWRKY40 and encoding gene and its application, can solve problems such as difficult to breed disease-resistant varieties

Active Publication Date: 2019-09-20
九圣禾种业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the difficulty in breeding cotton disease-resistant varieties lies in the acquisition of disease-resistant germplasm resources
At present, due to the lack of germplasm resources for disease resistance and immunity, it is difficult to effectively breed disease-resistant varieties using conventional breeding methods

Method used

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  • Plant disease-resistant protein GhWRKY40, coding gene and application thereof
  • Plant disease-resistant protein GhWRKY40, coding gene and application thereof
  • Plant disease-resistant protein GhWRKY40, coding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Experimental materials and reagents

[0052]Cotton material was China Cotton Research Institute 35, which was purchased from Seedling Company of Cotton Research Institute, Shanxi Academy of Agricultural Sciences; Escherichia coli 'DH5α' competent cells and Agrobacterium tumefaciens strain 'GV3101' were preserved by the State Key Laboratory of Plant Genomics; The strain (Verticillium dahliae, V.dahliae) 'V991' was donated by Jian Guiliang, a researcher at the Institute of Plant Protection, Chinese Academy of Agricultural Sciences; various restriction endonucleases and buffers were purchased from TaKaRa Company, and total plant RNA was isolated, extracted and purified by column The kit was purchased from Shanghai Sangon Bioengineering Co., Ltd., and the plasmid miniprep kit (upgraded spin column type) was purchased from Shanghai Jierui Bioengineering Co., Ltd.; TaqDNA polymerase, dNTPs, Taq Buffer, T4DNA ligase, PCR Purification recovery kit, PCR gel cutting recovery k...

Embodiment 2

[0074] 1. Experimental materials and reagents

[0075] With embodiment 1.

[0076] 2. Experimental method and result analysis: Analysis of the resistance of GhWRKY40 gene silenced plants to Verticillium dahliae

[0077] (1) Cultivation of virus-induced gene silencing (VIGS) plants (1.1) Construction of virus-induced gene silencing vector pYL-156-GhWRKY40

[0078] The viral silencing expression vector pYL-156 was added by Professor Liu Yule from Tsinghua University. Firstly, PCR technology was used to clone the target fragment of GhWRKY40 gene, and the primer used for amplification was VGhWRKY40-F: 5'-CG GGATCC GAATACTGAGAAAGAACTTAGTCCTAC-3'; VGhWRKY40-R:5'-GG GGTACC AACCGGAGTCGAACCAACAG-3' (the underlines are the restriction sites of BamH I and Kpn I, respectively). Digest the pYL-156 vector and the amplified product with restriction endonucleases BamH I and Kpn I, and then perform a ligation reaction to insert the fragment between the BamH I and Kpn I sites of the pYL-1...

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Abstract

The invention discloses a protein, the name of which is GhWRKY40, which is a protein shown in (a) or (b) or (c): (a) a protein with the amino acid sequence shown in sequence 2 in a sequence table; (b) a fusion protein obtained by connecting tags at the N end or / and the C end of the protein shown in the sequence 2 in the sequence table; and a protein with the function or activity of GhWRKY40 transcription factors which is obtained by deriving the protein shown in (a) or (b) through substitution, deletion or / and insertion of one or more amino acids. The gene has the advantages that the GhWRKY40 regulates transcription expression of specific genes, causes physiological and biochemical changes of plants, increases resistance of plants to pathogen invasion, so as to protect growth and development of plants, and has important significance in application of improving response of plants to pathogens.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, and in particular relates to a plant disease-resistant protein GhWRKY40, an encoding gene and an application thereof. Background technique [0002] Cotton belongs to the genus Gossypium in the family Malvaceae, and there are 4 cultivated species, including Upland Cotton, Sea Island Cotton, Grass Cotton and Asian Cotton, among which Upland Cotton is the main species. Cotton is an important economic crop, and its fiber is an important raw material for the textile industry. Cottonseed oil is one of the main sources of food and chemical oils. Cottonseed meal is a protein additive component of ruminant feed such as cattle and sheep. However, the yield and quality of cotton are affected by various biotic and abiotic stresses, among which cotton verticillium wilt seriously restricts the production of cotton. Verticillium wilt is a soil-borne vascular disease caused by Verticillium dahliae, which ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/60
CPCC07K14/415C12N15/8282
Inventor 吴家和刘建芬胡保民陈爱民
Owner 九圣禾种业股份有限公司
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