Plant stress resistance related protein TaWrky48 and coding gene and application thereof
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A stress resistance and encoding technology, applied in the direction of plant gene improvement, application, plant peptides, etc.
Inactive Publication Date: 2015-11-11
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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Embodiment 1
[0088] Embodiment 1, the cloning of wheat TaWrky48 gene
[0089] 1. Acquisition of cDNA
[0090] The RNA of wheat (Chinese Spring) was extracted, and cDNA was obtained by reverse transcription.
[0091] 2. PCR amplification
[0092] Using the cDNA obtained in step 1 as a template, PCR amplification was performed using TaWrky48F and TaWrky48R primers to obtain PCR amplification products. The primer sequences are as follows:
[0093] TaWrky48F: 5'-CTGACCACCGCCGTTTGA-3' (SEQ ID NO: 3);
[0094] TaWrky48R: 5'-ACAACAGGGAGCGGAGAAG-3' (SEQ ID NO: 4).
[0095] 3. Acquisition of TaWrky48 gene
[0096] Agarose gel electrophoresis detection of the PCR amplification product obtained in step 2 and sequencing: the results showed that the PCR amplification obtained an amplified fragment with a length of 414bp, which was named as TaWrky48 gene, and the nucleotide sequence of TaWrky48 gene is as in the sequence listing As shown in sequence 1, sequence 1 in the sequence listing consists o...
Embodiment 2
[0097] Example 2, the acquisition of trans-TaWrky48 rice and its stress resistance analysis
[0098] 1. Obtaining of rice transgenic for TaWrky48
[0099] 1. Acquisition of pCUbi1390-TaWrky48
[0100] (1) Acquisition of TaWrky48 gene
[0101] The RNA of wheat (Chinese Spring) was extracted, and cDNA was obtained by reverse transcription. The obtained cDNA was used as a template, and TaWrky48F and TaWrky48R primers were used for PCR amplification to obtain a PCR amplification product, namely the TaWrky48 gene. The primer sequences are as follows:
[0102] TaWrky48F: 5'-CTGACCACCGCCGTTTGA-3' (SEQ ID NO: 3);
[0103] TaWrky48R: 5'-ACAACAGGGAGCGGAGAAG-3' (SEQ ID NO: 4).
[0104] (2) the TaWrky48 gene obtained in step (1) and the pDONRTM / Zeo vector ( BP II (Invitrogen)) for BP reaction to obtain the entry vector.
[0105] (3) Transform the entry vector obtained in step (2) into E.coliDH5α cells by heat shock method, and the obtained clone is an entry clone, and the plasmi...
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Abstract
The invention discloses a plant stress resistance related protein TaWrky48 and a coding gene and application thereof. The plant stress resistance related protein provided by the invention is a protein of a), b) or c): a) protein with an amino acid sequence which is as shown by sequence 2 in a sequence table; b) fused protein which is obtained by connecting a label to the N end and/or C end of the protein with the amino acid sequence which is as shown by sequence 2 in the sequence table; c) protein which has the same functions and is obtained by substituting and/or deleting and/or adding one or a plurality of amino acid residues to the amino acid sequence which is as shown by sequence 2 in the sequence table. As proved by tests, the drought resistance of trans-TaWrky48 rice obtained by introducing the coding gene of the plant stress resistance related protein TaWrky48 disclosed by the invention to wild rice is obviously higher than that of the wild rice, and it has important theoretical and practical significance to the cultivation of new stress-resistant rice varieties, especially the cultivation of drought-resistant rice.
Description
technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a protein TaWrky48 related to plant stress resistance, its coding gene and application. Background technique [0002] Drought and salinity in the natural environment have an important impact on plant growth and development. In severe cases, it will cause large-scale crop yield reduction. Cultivating stress-resistant crops is one of the main goals of the planting industry. At present, it is an effective method to obtain stress-tolerant crop varieties through genetic engineering breeding. The most critical technical bottleneck in this method is the screening and function discovery of effective stress resistance genes. [0003] Transcription factors play an important role in the process of plant stress response. They can activate the expression of plant stress-related genes and the synthesis of different functional proteins, causing various physiological and biochemical reac...
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