Wheat salt tolerance gene TaCHP and use thereof

A wheat salt tolerance gene and gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, to achieve the effect of improving salt tolerance

Inactive Publication Date: 2009-07-15
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no report about the role of CHP zinc finger protein gene in the process of plant salt tolerance

Method used

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  • Wheat salt tolerance gene TaCHP and use thereof
  • Wheat salt tolerance gene TaCHP and use thereof
  • Wheat salt tolerance gene TaCHP and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Cloning of embodiment 1, TaCHP

[0040] 1.1 Extraction of wheat Total RNA

[0041] 1. Put the tissue material into a liquid nitrogen pre-cooled mortar, and fully grind it into powder in liquid nitrogen;

[0042] 2. After the liquid nitrogen has evaporated, transfer it to a 2ml centrifuge tube immediately, add about 1ml of Invitrogen’s TRIzol extract for every 100mg of material, after melting, repeatedly suck and blow with a sample gun, shake and mix the sample vigorously, and make the sample Fully lyse and place at room temperature for 5 minutes;

[0043] 3. Add 0.2ml of chloroform (chloroform), vigorously shake and mix for 15 seconds, and place at room temperature for 10 minutes;

[0044] 4.4°C, centrifuge at 12000rpm for 15 minutes;

[0045] 5. Use a pipette to carefully suck out the upper aqueous phase, add it to a new 1.5ml centrifuge tube, add 500μl of isopropanol (1:1 volume), mix well, and settling at -20°C for 30min or overnight;

[0046] 6.4°C, centrifuge a...

Embodiment 2

[0194] Embodiment 2, prokaryotic expression analysis

[0195] 2.1 Construction of prokaryotic expression vector

[0196] The expression vector used was pET24a, and the recipient strain was DE3. Hind III and EcoR I were used to perform double enzyme digestion on pET24a and the pMD18-T vector containing the target gene, respectively, and recover the large fragment of the vector and the small fragment of the target gene, and transform Escherichia coli DH10B competent cells after ligation with T4DNA ligase to identify the recombinant The prokaryotic expression vector with the target gene was obtained after the offspring, and then the recipient strain DE3 was transformed to be competent for protein expression.

[0197] 1. Digestion of pET24a and pMD18-T

[0198] The plasmids of pET24a and pMD18-T were extracted by alkaline lysis method, and 10ul of each was taken for HindIII and EcoRI double enzyme digestion. The plasmid extraction method is as above, and the plasmid double enzy...

Embodiment 3

[0271] Embodiment 3, tissue in situ hybridization

[0272] Tissue in situ hybridization was performed according to the method of Detection of mRNA intissue sections using DIG-labeled RNA from Roche Company. see results Image 6 .

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PUM

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Abstract

The invention discloses a salt tolerant gene of wheat, namely CHP zinc finger protein gene TaCHP of wheat and a plant expression vector containing the gene TaCHP. The invention also discloses an application of the gene TaCHP to culturing a salt tolerant plant, in particular culturing the wheat. Experiments prove that the salt tolerant ability of the transgenic plants of the invention is improved obviously.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to a salt-tolerant gene-zinc finger protein gene TaCHP and its application. Background technique [0002] Soil salinization seriously affects crop yields. Especially with the development of industry, soil salinization is becoming more and more serious, which has become a social problem of global concern. my country has a large population, and soil salinization is more serious, which has become an important factor restricting my country's economic and social development. Therefore, in addition to alleviating soil salinization, cultivating new varieties of salt-tolerant crops has become a very urgent task at present. [0003] It is a technology with broad application prospects to use transgenic improved plant technology to transfer new traits into high-biomass plants to develop new high-efficiency transgenic plant varieties for planting in saline-alkali la...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 夏光敏李翠玲赵欣
Owner SHANDONG UNIV
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