Protein related to salt tolerance, coding gene thereof and application thereof

A technology that encodes genes and proteins, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as slow growth, shedding, death, etc., and achieve the effect of improving salt tolerance and increasing yield

Active Publication Date: 2010-06-23
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most plants, especially crops, are sensitive to salt stress. When they grow in saline-alkali land, due to the stress of saline-alkali, they grow slowly, and their l

Method used

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  • Protein related to salt tolerance, coding gene thereof and application thereof
  • Protein related to salt tolerance, coding gene thereof and application thereof
  • Protein related to salt tolerance, coding gene thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. Screening of the salt-tolerance-related protein gene ThST3 and its cDNA cloning

[0045] 1. Screening of ThST3

[0046] The seedlings of Thellugiella halophila were treated with 200mM NaCl, the mRNA of the treated Thellugiella halophila was extracted and reverse-transcribed into cDNA, the cDNA library of Thellugiella halophila was constructed on the binary vector pGreen-SA, and the recombinant vector was used to transform Pseudomonas halophila. southern thaliana. At the same time, the Arabidopsis thaliana was transformed with the empty vector pGreen-GFP, and the obtained Arabidopsis was used as a control. The transgenic Arabidopsis was treated with 200mM NaCl, and the salt-tolerant strain was screened. It was found that strain No.0003 was more salt-tolerant than the control, and it was named ThST3 (Salt tolerant 3).

[0047] Genomic DNA of ThST3 strain Arabidopsis thaliana was extracted, the primers on the vector pGreen were used to amplify the insert fragm...

Embodiment 2

[0053] Embodiment 2, the acquisition of transgenic ThST3 gene plant and identification of stress tolerance

[0054] 1. Obtaining of ThST3 gene transgenic plants

[0055] The pTE-ThST3 vector was digested with XbaI and SacI, and the digested product was connected to the back of the 35S promoter of the binary vector pCAMBIA1300-221 (CAMBIA, Canberra, Australia) after the same digestion, and then the gene encoding the GFP fluorescent protein The recombinant plasmid was introduced to construct the sense expression vector pCAMBIA1300-221-35S-ThST3.

[0056] The sense expression vector was transformed into wild-type Arabidopsis (Col-0) by Agrobacterium-mediated vacuum infiltration. Screening was performed on an MS plate with a kanamycin concentration of 50 μg / ml, and 17 positive strains were obtained. The DNA of transgenic plants was extracted for PCR identification, and the results showed that all 17 transgenic lines were overexpressed lines of ThST3 gene. T 0 Transgenic Arabid...

Embodiment 3

[0082] Example 3, Long-distance transport activity of ThST3 protein and its application in cultivating salt-tolerant plants

[0083] 1. Long-distance transport activity of ThST3 protein

[0084] To identify whether ThST3 can carry out long-distance transport, Arabidopsis grafting experiment was carried out. Specific steps are as follows:

[0085] Get the Col-0 that grows 7d on the medium and the T from embodiment 2 3 For transgenic plants of generation 2.20, the seedlings were quickly cut horizontally from the middle of the cotyledon hypocotyl with a sharp blade, and then the breakpoints of the rootstock and the scion were quickly aligned, so that the cut surfaces of the scion and the rootstock matched as much as possible, and then melted low-melting point agar was dripped on it. After the agarose is cooled and solidified, place the plate vertically and continue to cultivate under the light. The entire operation process was carried out under a microscope and under sterile c...

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Abstract

The present invention discloses a protein related to salt tolerance, a coding gene thereof and an application thereof. The protein provided refers to the following protein (a) or the protein (b). The protein (a) consists of amino acid sequences shown in sequence 1 of a sequence table. The protein (b) related to plant salt tolerance is derived from the sequence 1 in which the amino acid sequences of the sequence 1 are replaced by one or several amino acid residues, and/or deleted, and/or added. The present invention also provides a coding gene of the protein. A transgenic plant of improved salt tolerance can be obtained by leading the coding gene of the protein into the plant, and novel plants of improved salt tolerance can be obtained by grafting the transgenic plant to other plants. The present invention has great significance in the cultivation of salt tolerant plants.

Description

technical field [0001] The invention relates to a salt-tolerance-related protein, its coding gene and application. Background technique [0002] At least 20% of the arable land and more than 40% of the irrigated arable land in the world are affected by salt damage to varying degrees. Soil salinization is a common concern in the world today. Salt stress is one of the main limiting factors for plant growth and development. Most plants, especially crops, are sensitive to salt stress. When they grow in saline-alkali land, due to the stress of saline-alkali, they grow slowly, and their leaves often turn yellow, die, and fall off, seriously affecting photosynthesis, and sometimes even the whole plant withers. death, resulting in reduced crop yields. Environmental stresses such as high salinity have greatly limited the increase in global food production. Therefore, understanding the response mechanism of plants to salt stress and improving plant salt resistance are important foun...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H1/00A01G1/00
Inventor 谢旗李刚张华伟夏然张译月
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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