Protein MtCBF4 associated with stress tolerance of plants, encoding gene and application thereof

A stress tolerance, gene technology, applied in plant genetic improvement, botanical equipment and methods, applications, etc., can solve problems such as significant differences in expression patterns

Inactive Publication Date: 2010-10-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although many salt-tolerance-related genes have been obtained, most of these genes come from the model organism Arabidopsis. Since the expression patterns of salt-tolerance genes are significantly different among different species, the salt-tolerance genes of Arabidopsis are transformed into other plants. These plants must be able to have a salt-tolerant phenotype, so it is necessary to study the salt-tolerant mechanism of various plants in depth and dig out salt-tolerant genes

Method used

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  • Protein MtCBF4 associated with stress tolerance of plants, encoding gene and application thereof
  • Protein MtCBF4 associated with stress tolerance of plants, encoding gene and application thereof
  • Protein MtCBF4 associated with stress tolerance of plants, encoding gene and application thereof

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Embodiment 1

[0043] Embodiment 1, the discovery of MtCBF4 gene

[0044] Establish a perfect alfalfa seed germination and seedling salt stress gene expression profile of truncated alfalfa through two microarray experiments, and perform cluster analysis and GO annotation analysis on genes significantly induced by salt stress, and perform GeneBins on the probes of interest Horizontal pathway classification analysis basically constructed alfalfa salt tolerance regulatory network framework. On this basis, a salt stress expression database of alfalfa truncation was established. A transcription factor whose expression was significantly up-regulated after salt induction was found, and the expression of this transcription factor was up-regulated by 64 times after induction with 200mM NaCL for 5 hours.

[0045] The protein composed of amino acid residues shown in Sequence 1 is named MtCBF4 protein, and the coding gene of MtCBF4 protein is named MtCBF4 gene, and its coding region is shown in the 11t...

Embodiment 2

[0046] Embodiment 2, the acquisition of transgenic plants and the identification of stress tolerance

[0047] 1. Cloning of MtCBF4 gene

[0048] Specific primer pairs were synthesized by Invitrogen.

[0049] MtCBF4_5': 5'-TAC CAT GGA CAT GTT TAC TAT GAA TCA ATT-3';

[0050] MtCBF4_3': 5'-ATA CTA GTT TAA AAT GAG TAA CTC CAC A-3'.

[0051] Using the truncated alfalfa cDNA as a template, PCR amplification was carried out with specific primers. The PCR amplification product was inserted into the pMD18-T simple vector (purchased from TaKaRa Bioengineering Company) to obtain the recombinant vector pMD18-Tsimple-MtCBF4. After sequencing verification, the PCR amplification product has the nucleotide (MtCBF4 gene) shown in sequence 2 in the sequence listing.

[0052] 2. Construction of recombinant expression vector (p1302-MtCBF4)

[0053] See the build process figure 1 .

[0054] 1. Digest the vector pMD18-T simple-MtCBF4 with restriction enzymes Nco I and Spe I, and reclaim the...

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Abstract

The invention discloses a protein MtCBF4 associated with the stress tolerance of plants, an encoding gene and application thereof. The protein provided by the invention is a protein as the following (a) or (b): (a) the protein consisting of an amino acid sequence shown by a sequence 1 in a sequence table; and (b) the protein which is prepared by carrying out substitution and/or deletion and/or addition of one or more amino acid residues on the amino acid sequence in the sequence 1, is associated with the stress tolerance of plants and is derived from the sequence 1. The MtCBF4 gene participates in salt stress and drought stress response reactions of medicago truncatula. A transgenic plant, of which the stress tolerance is obviously higher than a starting plant, can be obtained by introducing the MtCBF4 gene into the starting plant. The invention has important value for cultivation of transgenic leguminous crops with salt tolerance and drought tolerance.

Description

technical field [0001] The invention relates to a plant stress tolerance related protein MtCBF4 and its coding gene and application. Background technique [0002] As one of the main negative factors affecting grain yield and quality in the world, salt stress can cause harm to many crops. Under the condition of salt stress, the growth, development and biomass of crops will be greatly affected, especially the yield reduction of some crops that depend on irrigation is more obvious. More than one-fifth of the cultivated land in the world is threatened by salt stress, and in the context of global population growth and increasingly serious environmental pollution, the problem of salinization of land has become more prominent and has become a key to normal social and economic development. resistance. [0003] The adaptation of plants to salt stress in the natural environment is a complex process, which depends on the cascade activation of a series of molecular networks, namely: t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 董江丽王涛申小叶张运芹
Owner CHINA AGRI UNIV
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