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Plant stress tolerance associated protein GmNF-YA17, and encoding gene and application thereof

A stress tolerance and plant technology, applied in the direction of plant genetic improvement, botany equipment and methods, applications, etc., to achieve the effect of improving drought resistance

Inactive Publication Date: 2015-09-09
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are few reports on the functions of NF-Y transcription factors in plants, all of which play an important role in drought stress (Nelson et al, 2007)

Method used

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  • Plant stress tolerance associated protein GmNF-YA17, and encoding gene and application thereof
  • Plant stress tolerance associated protein GmNF-YA17, and encoding gene and application thereof
  • Plant stress tolerance associated protein GmNF-YA17, and encoding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, the acquisition of gene GmNF-YA17

[0063] 1. Acquisition of the gene GmNF-YA17

[0064] Soybean Tiefeng No. 8 grown in sandy soil for about 20 days (National Germplasm Bank, No. ZM242; recorded in the following documents: Wang Caijie, Sun Shi, Wu Baomei, Changru Town, Han Tianfu. Since the 1940s, a large area of ​​China Pedigree Analysis of Soybean Varieties. Chinese Journal of Oil Crops. 2013, 35(3): 246-252, available to the public from the Institute of Crop Science, Chinese Academy of Agricultural Sciences) Seedlings at the four-leaf stage were drought-treated for 2 hours, quick-frozen with liquid nitrogen,- Store at 80°C for later use. mRNA was isolated using Quikprep Micro mRNA Purification Kit (Pharmacia). First-strand cDNA synthesis was performed with Reverse Transcriptase XL (AMV). The ds cDNA was synthesized by SMART method, and the PCR products were detected by 1.0% agarose gel electrophoresis.

[0065] Sequence 2 in the full-length sequence...

Embodiment 2

[0125] Example 2, Application of GmNF-YA17 as a transcription factor

[0126] The main principle of using the yeast one-hybrid system to prove the activation properties of transcription factors is as follows: construct the CCAAT cis-acting element and the mutant CCAAT cis-acting element to the upstream of the basic promoter Pmin (minimal promoter) of the pHISi-1 vector and the pLacZi vector, respectively , The reporter genes (His3, LacZ and URA3) were connected downstream of the Pmin promoter. When the expression vector YEP-GAP (without activation function) connected with the target gene encoding the transcription factor is transformed into the yeast cells connected with the CCAAT cis-acting element and the mutant CCAAT cis-acting element, if the mutant CCAAT The reporter gene in yeast cells with cis-acting elements cannot be expressed, but the reporter gene in yeast cells with specific CCAAT cis-acting elements can be expressed, indicating that the transcription factor can bi...

Embodiment 3

[0178] Embodiment 3, the application of GmNF-YA17 in improving the stress tolerance of plants

[0179] 1. Obtaining transgenic GmNF-YA17 Arabidopsis

[0180] 1. Construction of recombinant vector

[0181] 1) Cloning of GmNF-YA17 gene

[0182] A primer pair (GmNF-YA17-121F and GmNF-YA17-121R) was designed according to the sequence of the GmNF-YA17 gene.

[0183] GmNF-YA17-121F: 5'-TCCCCCGGGATGCAGTCCAAGTCTGAAAC-3'

[0184] GmNF-YA17-121R: 5'-TACGAGCTCTCAATTGTGGTTCAGCTGCT-3'

[0185] Glycine max L. (Tiefeng 8) cDNA was used as a template, and GmNF-YA17-121F and GmNF-YA17-121R were used for PCR amplification to obtain a PCR product (GmNF-YA17 gene) with a size of about 1Kb. .

[0186] The above PCR product was recovered.

[0187] 2) Construction of recombinant vector

[0188] ① Digest the PCR product recovered in step 1 with restriction enzymes SmaI and SacI, and recover the 1029bp digested product;

[0189] ② Digest pBI121 (purchased by Clontech) with restriction enzymes ...

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Abstract

Plant stress tolerance associated protein GmNF-YA17, and an encoding gene and an application thereof. The protein is a protein (a) composed of an amino acid sequence represented by sequence 1 in a sequence table, or a protein (b) obtained by substituting and / or deleting and / or adding one or more amino acid residues to the protein (a), associating with the plant stress tolerance and derived from the sequence 1. Experiments prove that gene GmNF-YA17 can be obtained through cloning from soybeans, is expressed under induction of drought, salt, ABA and ethylene, positions the coded protein to a nucleus, and can specifically regulate the transcription expression of gene containing a CCAAT-box cis-element. The GmNF-YA17 can improve the drought resistance of plants, provides a foundation for artificial control of expression of stress resistance and stress tolerance associated genes, and plays a great role in the cultivation of stress resistance and stress tolerance reinforced plant.

Description

Technical field [0001] The invention relates to the field of biotechnology, and in particular to a plant stress tolerance-related protein GmNF-YA17 and its encoding gene and application. Background technique [0002] Adverse stresses such as drought, high salinity and low temperature seriously restrict the growth and development of soybeans. Therefore, understanding the response and signal transduction mechanism of soybean to stress conditions and improving the stress resistance of soybean varieties have become one of the important tasks in soybean genetic research and variety improvement. [0003] Under adverse stress, plants will produce a series of responses, accompanied by many physiological, biochemical and developmental changes. Clarifying the response mechanism of plants to stress will provide scientific evidence for the research and application of stress-resistant genetic engineering. At present, research on plant stress resistance has gradually deepened to the cel...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N15/82A01H5/00
CPCC07K14/415C12N15/8273
Inventor 徐兆师马有志冯志娟郑炜君陈明李连城
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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