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Application of stress-tolerant relative protein MtMYB5 of plant in cultivating stress-tolerant plant

A stress-tolerant plant and protein technology, applied in the field of genetic engineering, can solve problems such as unsatisfactory effects

Inactive Publication Date: 2011-10-05
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In many studies on plant stress resistance, scientists have cloned many stress-related functional genes from different sources and transformed them into plants, but the results are not satisfactory

Method used

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  • Application of stress-tolerant relative protein MtMYB5 of plant in cultivating stress-tolerant plant
  • Application of stress-tolerant relative protein MtMYB5 of plant in cultivating stress-tolerant plant
  • Application of stress-tolerant relative protein MtMYB5 of plant in cultivating stress-tolerant plant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Cloning of embodiment 1, MtMYB5 gene

[0030] 1. Acquisition of MtMYB5 gene

[0031] The gene whose expression was significantly up-regulated was found in chip screening of alfalfa truncated A17 after salt stress. In order to study the function of this gene in abiotic stress, 4-week-old seedlings of alfalfa truncated A17 were poured with 250 mM NaCl aqueous solution for one hour. RNA was extracted, reverse-transcribed to obtain cDNA, and the cDNA of truncated alfalfa A17 was used as a template, and MtMYB5___5' and MtMYB5___3' were used as primers to carry out PCR amplification to obtain a PCR product, which was inserted into the pMD18 T-simple vector (purchased from TaKaRa Bioengineering Company), obtained the recombinant vector, sequenced, and the result was that the PCR product had the nucleotide sequence 1 in the sequence listing, and the gene of the PCR product was named MtMYB5, and the coding region of the gene was sequence 1 to 5 in the sequence listing. 1-837 at...

Embodiment 2

[0042] Example 2, Obtaining and Functional Research of Transforming MtMYB5 Arabidopsis

[0043] 1. Construction of pCAMBIA1302-MtMYB5 constitutive high-efficiency expression vector

[0044] Invitrogen synthesized specific primer pairs containing BglII and Spe I restriction sites respectively:

[0045] 5'-AGATCTTATGGATACTAATTACAAAACCAATA-3' (SEQ ID NO: 5)

[0046] 5'-ACTAGTTCATAAATCATCAGCCAATTGTTGC-3' (SEQ ID NO: 6)

[0047] Using pMD18 T-simple-MtMYB5 as a template, using the DNA molecule shown in Sequence 5 and the DNA molecule shown in Sequence 6 as primers, amplified to obtain a fragment containing the restriction sites of BglII and Spe I, which is the same enzyme The cut vector pCAMBIA1302 (Center for the Application of Molecular Biology to International Agriculture, www.cambia.org) was ligated, and the ligated product was transformed into Escherichia coli to obtain a transformant, and the plasmid of the transformant was extracted and sequenced. The results showed that t...

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Abstract

The invention discloses an application of a stress-tolerant relative protein MtMYB5 of a plant in cultivating a stress-tolerant plant. According to the application of the MtMYB5 protein in cultivating the stress-tolerant plant, an amino acid sequence of the MtMYB5 protein is a sequence 2 in a sequence table; and the application is as follows: an encoding gene of the MtMYB5 protein is introduced into a target plant to obtain a transgenic plant with stress tolerance higher than that of the target plant. According to the invention, experiments prove that a transcription factor MtMYB5 outstandingly increased in expression level after being induced by salt is screened, and important value is reflected on cultivating salt-tolerant drought-tolerant transgenic leguminous crops by introducing the gene into leguminous crops, i.e. cash crop alfalfa and the like.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the application of plant stress tolerance-related protein MtMYB5 in cultivating stress-tolerant plants. Background technique [0002] Drought, low temperature, soil salinization, diseases and pests are serious problems affecting agricultural production. In many studies on plant stress resistance, scientists have cloned many stress-related functional genes from different sources and transformed them into plants, but the results were not satisfactory. Plant stress resistance traits are complex traits controlled by multiple genes. The tolerance of plants to high salinity, drought and low temperature often does not depend on a single factor, and its traits are affected by many factors. [0003] Transcription factors, also known as trans-acting factors, regulate the transcription of downstream genes by binding to specific DNA elements. Some transcription factors act as transcript...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00
Inventor 董江丽王涛胡晓娜张运芹
Owner CHINA AGRI UNIV