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Protein capable of enhancing plant drought resistance and coding gene and application thereof

A gene and plant technology, which is applied to a protein that enhances plant drought resistance and its coding gene and application fields, can solve the problem of not bred drought-resistant corn varieties, etc.

Inactive Publication Date: 2019-12-24
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the world has not bred a genetically modified "drought-resistant maize variety"

Method used

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  • Protein capable of enhancing plant drought resistance and coding gene and application thereof
  • Protein capable of enhancing plant drought resistance and coding gene and application thereof
  • Protein capable of enhancing plant drought resistance and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Preparation of transgenic drought-resistant corn

[0025] The corn seed material is:

[0026] (1) With reference to the sequence of SEQ ID NO.1, use software to design primers for cloning the gene, and clone the full-length cDNA gene from the maize cDNA library;

[0027] (2) The cDNA sequence of the target gene was cloned into the commercial plasmid pGSA1252 at the Nco1 / BamH1 restriction site downstream of the 3×CaMV35S strong promoter to obtain the recombinant plasmid pGSA1252::RA33G4; the commercial plasmid pGSA1252 contains a chloramphenicol-resistant Gene and selection marker gene Bar for herbicide resistance Basta;

[0028] (3) preparing the Agrobacterium tumefaciens introduced into the plasmid into a bacterial liquid with an OD600 of 0.5 and containing 100 μM of acetosyringone;

[0029] (4) Slightly wound the mature embryo of corn with the tip of a scalpel, then sterilize it with 75% alcohol, then transfer the seeds into the bacterial solution of step ...

Embodiment 2

[0033] Example 2 Molecular identification of transgenic maize plant material with high drought resistance

[0034] 1. Identification by PCR method

[0035] The genomic DNA of the plant material was extracted, and the fragments of the Basta gene and 35S-RA33G4 gene were amplified by PCR, wherein the primer sequences for amplifying the Basta gene were: Bar-F 5'-GCACCATCGTCAACCACTACATCG-3' and Bar-R 5'-AAATCTCGGTGACGGGCAGGAC3 ', the primer sequences for amplifying the 35S-RA33G4 gene are: 35S–F5'-CGTCTTCAAAGCAAGTGGATTG-3' and RA33G4–R 5'-GCGGTACCGTGACAGATGATGCATGGGG-3'. The 35S-RA33G4 gene primer pair amplified the partial segments of the 35S and RA33G4 genes. The specific PCR process and conditions were carried out according to conventional methods, and the PCR products were subjected to agarose gel electrophoresis ( figure 1 ), figure 1 M is 100bp marker; 1: positive plasmid; 2: non-transgenic plant; 3: unreverse-transcribed RNA; band, confirmed to be transgenic plants.

...

Embodiment 3

[0038]Example 3 Identification of the drought resistance function of the self-homogenous T4 generation of transgenic maize plants

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Abstract

The invention provides a gene capable of enhancing plant drought resistance, a protein encoded by the gene and application of the protein in acquirement of high-drought-resistance corn by using a transgenic technology. Transgenic corn plants are obtained by introducing the gene into corn through a recombinant eukaryotic expression vector taking chloramphenicol resistance and herbicide resistance Basta as selection markers. The invention provides the gene capable of enhancing plant drought resistance, the protein encoded by the gene and application of the protein in cultivation of high-drought-resistance corn, and the gene is introduced into corn plants through an agrobacterium tumefaciens-mediated method so as to obtain transgenic corn. Compared with an untransformed corn variety, the transgenic corn plants have relatively strong drought resistance.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a protein that can enhance the drought resistance of plants and its encoding gene and application. Background technique [0002] Maize (Oryza sativa L.) is an important food crop. Corn production is a water-intensive process, so corn production is very sensitive to water scarcity. The main corn planting periods in my country (April to September in Northeast China, March to October in Huanghuai Region, March to November in the Yangtze River Basin, and January to December in South China and Southwest China) often suffer from varying degrees of drought. According to preliminary estimates, the crop drought-affected area and the drought-affected area are roughly increasing at a rate of 4.088 million hectares and 2.090 million hectares per decade. According to the forecast of the Agricultural Policy Research Center of the Chinese Academy of Agricultural Sciences, my country w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/84A01H5/00A01H6/46
CPCC07K14/415C12N15/8205C12N15/8273
Inventor 李有志樊宪伟潘虹胡海洋吴振波
Owner GUANGXI UNIV
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