Protein related to plant heat resistance property and coding gene and application thereof

A technology that encodes genes and proteins, which can be applied in the fields of plant gene improvement, application, plant peptides, etc., and can solve the problems of less research and development.

Inactive Publication Date: 2009-11-25
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

And the overexpression of GASA4, a member of the GAST family in Arabidopsis, improves the h

Method used

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  • Protein related to plant heat resistance property and coding gene and application thereof
  • Protein related to plant heat resistance property and coding gene and application thereof
  • Protein related to plant heat resistance property and coding gene and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1, the cloning of TaGAST gene

[0044] (1) Electronic cloning of wheat TaGAST gene sequence

[0045] According to Affymetrix The probe sequence given by the wheat Genome array with the probe number "Ta.11162.1.S1_at" was compared by Blast on the NCBI (www.ncbi.nlm.nih.gov) website to obtain a highly homologous Wheat EST sequences, download these EST sequences, and use the sequence assembly program of DNAMAN software to splice the partially overlapping sequences in the results, and obtain the ORF splicing sequence with start codon and stop codon. Use tblastx in GeneBank to verify whether the amino acid sequence encoded by the fragment contains the predicted functional domain, design PCR-specific primers for the ORF, and perform amplification. The primer pair used was: 5'-GACCAGGATGAGCAAGCCAT-3' and 5'-GTGGAGTAGAGGTTGAAGAAGCC-3'.

[0046] (2) Cloning of wheat TaGAST gene promoter

[0047] The promoter of wheat TaGAST gene was cloned by using BD GenomeWalker...

Embodiment 2

[0051] Embodiment 2, the stress-induced expression pattern of TaGAST gene

[0052] 20% polyethylene glycol (PEG-6000) stress, 250 mM sodium chloride (NaCl) salt stress, 100 μM H 2 o 2 Oxidative stress treatment, and phytohormone ACC 200μM stress, ABA 100μM stress, MeJA 100μM stress, gibberellin (GA) stress and heat (HS) stress were used to treat wheat varieties TAM107 (Triticum aestivumL.) and Chinese Spring (Chinese Spring, ) of seedlings grown for ten days, and wheat leaves with different treatment times were taken, RNA was extracted and cDNA was reverse-transcribed, and primer 1: 5'-CCACCGTCTCAAGTCTCAAC-3' and primer 2: 5'-ACAGGCAGCAAACTCACTC-3' were used.

[0053] The temporal and spatial expression characteristics of TaGAST gene were detected by real-time fluorescent quantitative PCR. The specific methods of coercion treatment are as follows:

[0054] Heat treatment and gibberellin treatment: at the same time, transfer 2 wheat materials into ①40℃ light incubator for he...

Embodiment 3

[0065] Example 3, Phenotype and Physiological Identification of Wild Type and Transgenic Plants Under High Temperature Stress Treatment Conditions

[0066] 1. Obtaining of transgenic plants

[0067] (1) Construction of pCAMBIA-35S-TaGAST expression vector

[0068] Using the cDNA of the leaves of the wheat variety TAM107 (Triticum aestivum L.) as a template, using primers 3: 5'-GCTCTAGAATGGCGCAGCGGGATCACA-3' and 5'-GGGGTACCTCAGAACCTGACGAGCTTG-3', PCR amplification obtained the TaGAST coding sequence (sequence 1 in the sequence listing The nucleotide sequence from the 696th to the 992nd deoxynucleotide of the 5' end), the obtained TaGAST fragment was digested with restriction endonucleases XbaI and KpnI, and inserted into plasmid pCAMBIA-1300 (purchased from CAMBIA Company) Between the restriction endonuclease XbaI and the KpnI restriction site, obtain the expression vector pCAMBIA-TaGAST containing TaGAST coding region fragment; With restriction endonuclease HindIII and XbaI d...

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Abstract

The invention discloses a protein related to plant heat resistance property and coding gene and application thereof. The name of the protein related to plant heat resistance property provided by the invention is TaGAST (Triticum aestivum Gibberellin stimulated transcript), the protein is originated from the common wheat (Triticum aestivum L.), is the protein of the following 1) or 2): 1) the protein composed of amino acid sequence shown by the sequence 2 in the sequence table; 2) the protein derived from the 1) related to the plant heat resistance property with amino-acid residue sequence of the sequence in the sequence table by the substitution and/or deletion and/or adding of one or several amino-acid residues. The protein TaGAST related to plant heat resistance property and coding gene provided by the invention has important theory and actual meanings for cultivating new species such as crops, forests and grasses with improved stress resistance, can be used for the cultivating and the identifying of stress resistance plant species required by the farming and animal husbandry and ecologic environment harnessing.

Description

technical field [0001] The invention relates to a protein related to plant stress resistance and its coding gene and application, in particular to a protein related to plant heat resistance and its coding gene TaGAST and application. Background technique [0002] As the global population increases, the demand for food is increasing. As the main food crop in the world, wheat is a cool C 3 For crops, the optimum growth temperature is 23°C-25°C. High temperature reduces wheat yield by 10%-20% in the main wheat areas of my country, which is an important limiting factor in wheat production, especially the high temperature stress in the late growth period seriously affects the yield of wheat and quality, have brought great influence to people's life. Therefore, how to improve the heat tolerance of wheat, explore the mechanism of wheat heat tolerance, develop heat tolerance-related genes, and apply them to wheat breeding, so as to breed heat-tolerant varieties has become an import...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/00C12N15/11C12N15/82
Inventor 彭惠茹周春蕾孙其信倪中福姚颖垠梁荣奇解超杰刘志勇杜金昆秦丹丹
Owner CHINA AGRI UNIV
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