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A protein related to plant stress tolerance and its coding gene scmyb3r1 and application

A technology for encoding genes and plants, applied in the direction of plant gene improvement, application, plant peptides, etc.

Active Publication Date: 2018-03-02
CROP RES INST SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the characteristics of rye is mainly focused on phenotypic identification, map construction and gene positioning, and there are relatively few cloning and functional studies of excellent rye genes. Therefore, the in-depth excavation and application of excellent rye genes is of great importance Improving the stress resistance of plants is of great significance

Method used

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  • A protein related to plant stress tolerance and its coding gene scmyb3r1 and application
  • A protein related to plant stress tolerance and its coding gene scmyb3r1 and application
  • A protein related to plant stress tolerance and its coding gene scmyb3r1 and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, the acquisition of ScMTB3R1

[0046] 1. Acquisition of ScMTB3R1

[0047] 1. Treat the seedlings of rye AR132 grown for 2 weeks in a refrigerator at 4°C for 5 hours to obtain the treated seedlings; extract the total RNA of the treated seedlings, and construct a cDNA library according to the instructions of Bao Bio cDNA Library Construction Kit; use 12% agar Sugar gel electrophoresis detection, recovery and purification.

[0048] 2. Connect the purified cDNA to the NTT vector, transform E.coli strain DH5α, collect all the grown E. coli colonies, extract the mixed plasmids and transform yeast EGY48, and spread the auxotrophic plate (SD / His2 / Leu2), 30°C Culture for 2-4 days, and use NTT to screen for nuclear localized proteins.

[0049] 3. Extract yeast plasmids from the screened yeast clones and transform them into Escherichia coli. A batch of clones are obtained after sequencing. One of the proteins contains the MYB domain. The protein was compared at NCBI...

Embodiment 2

[0063] Example 2, Acquisition and Functional Analysis of Transgenic ScMTB3R1 Arabidopsis

[0064] 1. Construction of recombinant expression vector

[0065] 1. Extraction of RNA

[0066] The seedling stage rye AR132 was used as the experimental material, and the total RNA was extracted by the TRNzol method, and the qualified RNA was reverse-transcribed into cDNA with the reverse transcription kit of TaKaRa Company, and stored at -20°C for future use.

[0067] 2. PCR amplification

[0068] The cDNA obtained above was used as a template, and primers ScMYB3R1-F2 and ScMYB3R1-R2 were used for PCR amplification to obtain the target gene ScMYB3R1. The primer sequence is as follows: ScMYB3R1-F2: 5'-TCC CCCGGG ATGGGGGCCATGGCGGAGG-3' (the underlined part is the recognition site of Sma I);

[0069] ScMYB3R1-R2: 5'-GG ACTAGT TTAGGTTACATCCATGTTTGTCG-3' (the underlined part is the recognition site of Spe I).

[0070] PCR reaction system: 2×GC Buffer 25μl, dNTP Mix 4μl, upstream and ...

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Abstract

The invention discloses a plant stress tolerance correlated protein as well as an encoding gene ScMYB3R1 and applications thereof. The protein is a protein (a) or (b) shown as follows: a protein of which the amino acid sequence is shown in a sequence 2 in a sequence table; and (b) a protein which is obtained by carrying out substitution and / or deletion and / or addition on the amino acid sequence shown in the sequence 2 in the sequence table by using one or more amino acid residues and can improve the plant stress tolerance correlation. Experiments show that the protein provided by the invention has a function of improving the plant stress tolerance, and the protein has an important application value in the aspect of plant breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a protein related to plant stress tolerance, its coding gene ScMYB3R1 and its application. Background technique [0002] Temperature is one of the important environmental factors for plant growth, which limits the distribution, growth and yield of plants. When the temperature of the external environment drops to a certain level, the plant body will be injured or even die. Under the condition of low temperature stress, plants will naturally produce a series of physiological and biochemical reactions and induce the expression of a series of related genes. Different plants respond differently to low temperature, some plants show an increase in the expression level of original proteins, and some plants produce low temperature-induced proteins. [0003] The expression products of low temperature stress-induced genes can be divided into two categories according to their functi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 曹新有刘建军程敦公王灿国刘爱峰李豪圣宋健民刘成孙正娟赵振东
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI