Salicornia europaea SeVHA-A protein and encoding gene and application thereof

A technology that encodes genes and proteins, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as soil salinization exacerbated by cultivation measures

Inactive Publication Date: 2011-10-05
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Improper cultivation practices are still exacerbating soil salinization

Method used

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  • Salicornia europaea SeVHA-A protein and encoding gene and application thereof
  • Salicornia europaea SeVHA-A protein and encoding gene and application thereof
  • Salicornia europaea SeVHA-A protein and encoding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the preparation of gene and protein

[0054] 1. Gene preparation

[0055] 1. Cloning of Salicornia seVHA-A gene

[0056] (1) Acquisition of 3'RACE gene fragment

[0057] The total RNA of Salicornia salicornia leaves was extracted by Trizol mini-extraction method, and the first-strand cDNA was synthesized by a reverse transcription kit (Quanshi Jinbio). SeV-1F / 3'oligdT was amplified by PCR with primers to obtain the 3' terminal fragment of cDNA. The specific PCR reaction conditions are as follows: 94°C 5min; 2 cycles: 94°C 30sec, 68°C 30sec, 72°C 1min; 2 cycles: 94°C 30sec, 66°C 30sec dT=-2°C, 72°C 1min; 25 cycles: 94°C 30sec, 54°C 30sec, 72°C 1min; 72°C 10min.

[0058] Upstream degenerate primer SeV-1F: 5'-CCTCTA(A / T / G)CTC(G / C / T)ACTGGACAGCG-3'. The downstream primer is 3'oligdT.

[0059] The obtained PCR products were subjected to agarose gel electrophoresis, and DNA fragments with similar sizes to the expected bands were recovered and sequenced afte...

Embodiment 2

[0088] Embodiment 2, the application of gene

[0089] 1. Overexpression—transformation of wild-type Arabidopsis

[0090] 1. Recombinant expression vector

[0091] The plant expression vector pBI121 was disclosed in the document "Overexpression of Organellar and Cytosolic AtHSP90 in Arabidopsis thaliana Impairs Plant Tolerance to Oxidative Stress. Hongmiao Song, Pengxiang Fan, Yinxin Li. Plant Mol Biol Rep (2009) 27: 342-349." Publicly available from Institute of Botany, Chinese Academy of Sciences. The plant expression vector pBI121 used contained a Kana (kanamycin) selection marker, and SeVHA-A and AtVHA-A genes were respectively inserted for gene function verification in Arabidopsis thaliana.

[0092]The SeVHA-A gene obtained in Example 1 was inserted between the Xba I and Sma I restriction sites of the plant expression vector pBI121, and the resulting recombinant expression vector was denoted as pBI121-SeVHA-A; sequencing verification, the results were in pBI121-SeVHA- T...

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Abstract

The present invention discloses a Salicornia europaea SeVHA-A protein and an encoding gene and application thereof. The protein is selected from: a) a protein composed of the amino acid sequence shown in SEQ ID NO: 1; or b) a protein, which is associated with the plant adverse resistance and is derived from a) by the substitution and/or deletion and/or addition of the amino acid sequence shown inSEQ ID NO: 1 with one or more amino acid residues. Experiments prove that the gene of the Salicornia europaea SeVHA-A protein provided by the invention has a salt stress resisting function, therefore, the Salicornia europaea SeVHA-A protein provided by the invention has great significance and has broad application prospects in the fields of plant genetics and breeding.

Description

technical field [0001] The invention relates to Salicornia seVHA-A protein, its coding gene and application. Background technique [0002] According to statistics from the Food and Agriculture Organization of the United Nations in 2008, the saline soil in the world has exceeded 800 million hectares. Improper cultivation measures are still exacerbating soil salinization. Salt stress is the main abiotic stress faced by plants. It seriously affects the growth of plants and causes yield reduction. Therefore, it is especially important to improve the salt resistance of crops. [0003] The growth and development of plants are inseparable from material transport and energy conversion, and ATPase plays an important role in the two. The main function of V-ATPase (V-typeATPase, vacuolar ATPase) in plant cells is to use the energy generated by the hydrolysis of ATP to convert H + Pump into the vacuole, adjust the pH of the vacuole and form a transmembrane electrochemical potential g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16C12N15/55C12N15/11C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12N7/01A01H5/00
Inventor 李银心江萍
Owner INST OF BOTANY CHINESE ACAD OF SCI
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