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Peanut salt-tolerant associated gene Rab7 and application thereof to improvement of salt tolerance

A salt-tolerance, peanut technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., to achieve the effect of improving salt tolerance

Inactive Publication Date: 2012-07-25
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But it has not been seen in peanuts yet Rab7 Reports on Gene Cloning and Functional Research

Method used

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  • Peanut salt-tolerant associated gene Rab7 and application thereof to improvement of salt tolerance
  • Peanut salt-tolerant associated gene Rab7 and application thereof to improvement of salt tolerance
  • Peanut salt-tolerant associated gene Rab7 and application thereof to improvement of salt tolerance

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1: Peanut Rab7 Gene and its Application in Improving Salt Tolerance of Peanut

[0033] 1. Experimental materials

[0034] 1. Genes, strains and rice varieties

[0035] The present invention clones peanut Rab7 Gene( AhRab7 ), Escherichia coli DH5α was preserved by the genetic research laboratory of Qingdao Agricultural University, Agrobacterium tumefaciens strain EHA105 (purchased from Beijing Tianenze Gene Technology Co., Ltd.), and the recipient material of the transgene was peanut variety "Xuzhou 68-4" (Qingdao Provided by the Laboratory of Genetics, Agricultural University).

[0036] 2. Plant medium

[0037] The media used in peanut transformation are:

[0038] Basic medium: MS inorganic salt + B 5 Organic ingredients (referred to as MSB 5 ), add 3% sucrose, 0.8% agar, pH=5.8, and sterilize at 121℃, 105Kpa for 20min;

[0039] SIM induction medium: MSB 5 +5 mg / L BAP +1.5 mg / L 2,4-D;

[0040] SEM shoot elongation medium: MSB 5 +5 mg / L BAP.

[0...

Embodiment 2

[0069] Embodiment 2, peanut Rab7 Application of Genes in Improving Salt Tolerance of Escherichia coli

[0070] 1. Experimental materials

[0071] 1. Strains and rice varieties

[0072] Escherichia coli BL21 and Escherichia coli strain pET-28a were preserved by the Laboratory of Genetics, Qingdao Agricultural University.

[0073] 2. Plant medium

[0074] The medium used is LB medium.

[0075] 2. Experimental method

[0076] The present invention comprises following specific experimental steps:

[0077] 1, AhRab7 Construction of gene prokaryotic expression vector

[0078] to carry AhRab7 Genetic pUC18 recombined plasmid as template, amplified by PCR AhRab7 Gene, the primers used are:

[0079] P5 (5-CATATGATGCCTTCCAGAAGAAGAAC-3) ( Nde Ⅰ) (SEQ ID No: 8);

[0080] P6 (5-AAGCTTTCATCTCAGCATTCACAACCTGT-3) ( Hin dIII) (SEQ ID No: 9);

[0081] The PCR product was recovered and ligated with the cloning vector pUC18 under the action of T4 DNA ligase, and the ligated pro...

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Abstract

The invention provides a peanut salt-tolerant associated gene Rab7 (AhRab7) and an Rab7 (RabG3f) protein generated by encoding the gene. The amino acid sequence similarity of the protein to AtRabG3f in arabidopsis thaliana is 92 percent. After the gene is excessively expressed in a peanut, a transgenic plant of which the salt tolerance is obviously improved is obtained. After the gene is expressed in escherichia coli, the salt tolerance of a recombinant bacterium is obviously improved. The tolerance of the peanut and the escherichia coli to high-salt stress can be obviously improved through the over expression of the AhRab7 gene, and the normal growth and the economic character of the peanut are not obviously affected. The protein and the gene encoding the protein have an important theoretic and practical significance in study on the salt-tolerant mechanism of plants and the improvement of associated characters of the plants. The protein and the gene have an important effect in the improvement of salt-tolerant genetic engineering of the plants and have wide application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to genes related to peanut salt tolerance Rab7 ( AhRab7 ) and its application in improving salt tolerance. Background technique [0002] Peanut is an important oil crop and economic crop, which plays an important role in my country's agricultural production and even the entire national economy. Salt stress inhibited the growth of peanuts, resulting in wilting of plants, slow growth of pods, significant increase of small fruit, and reduced yield. At the same time, the quality of pods deteriorated and the commercial value decreased significantly. With the continuous deterioration of the environment, cultivating new varieties of crops with salt tolerance has become one of the main goals of the research of the majority of breeders. The current rapid development of genetic engineering technology provides a new way for plant genetic improvement, genetic transformation using ge...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/63C12N1/21C07K14/415A01H5/00
Inventor 隋炯明李瑞王晶珊乔利仙郑春花郭宝太
Owner QINGDAO AGRI UNIV
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