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A transcription factor pwnac2 related to plant stress tolerance and its coding gene and application
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A stress-tolerant, plant-based technology, applied in the fields of plant gene improvement, application, plant peptides, etc., can solve the problem of less research reports on NAC gene function
Active Publication Date: 2021-04-16
BEIJING FORESTRY UNIVERSITY
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At present, important progress has been made in the research on the relationship between NAC gene and plant stress resistance in model plants such as Arabidopsis and rice, but there are few reports on the function of NAC gene in woody plants
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Embodiment 1
[0080] Example 1. Acquisition of transcription factor PwNAC2 and its coding gene
[0081] 1. Acquisition of the transcription factor PwNAC2 and its coding gene
[0082] The Gateway method was used to construct the Cinnamon cDNA library, which was completed by Yingwei Jieji (Shanghai) Company. Using the EST sequence of PwNAC2 of P. japonicus as template, 5'-CCCAATCATCCAACCTTAGGCT-3' and 5'-CAACTGGGCCTCTGCATTC-3' primers were used for PCR amplification to obtain 5'RACE-PCR; -CTGCCAGGAAACAGCTATGAC-3'primers were used for PCR amplification to obtain 3'RACE-PCR, and the RACE-PCR product was spliced with EST sequence to obtain the full-length cDNA sequence, which was then sequenced.
[0083] Sequencing results show that: the cDNA nucleotide sequence is sequence 1 in the sequence table, consisting of 1164 nucleotides, compared with the sequence in Genbank, it is determined that it belongs to the NAC transcription factor of C. PwNAC2, the protein encoded by PwNAC2 is named PwNAC2,...
Embodiment 2
[0087] Example 2, Application of PwNAC2 Gene in Improving Plant Stress Tolerance
[0088] 1. Construction of transgenic Arabidopsis thaliana
[0089] 1. Acquisition of PwNAC2 gene
[0090] Prepare the following primer pairs:
[0091] Primer 1: 5'-CAACATGGGGAGACCGAATGC-3';
[0092] Primer 2: 5'-TCAATCCCTCTAATAAGAA-3'.
[0093] Using the cDNA of Pseudomonas japonica as a template, PCR amplification was carried out with the above primers 1 and 2 to obtain a PCR product with a size of 1164bp, which was then sequenced.
[0094] Sequencing results show that the nucleotide sequence of the PCR product is sequence 1 in the sequence listing, which can encode the protein PwNAC2 shown in sequence 2 in the sequence listing.
[0095] 2. Acquisition of recombinant expression vector
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Abstract
The invention discloses a transcription factor PwNAC2 related to plant stress tolerance, its coding gene and application. The protein provided by the present invention is a protein as follows a) or b) or c) or d): a) the amino acid sequence is the protein shown in sequence 2; b) at the N-terminal and / or C-terminal of the protein shown in sequence 2 A fusion protein obtained by linking tags; c) a protein with the same function obtained by substituting and / or deleting and / or adding the amino acid sequence shown in Sequence 2 by one or several amino acid residues; d) the same function as shown in Sequence 2 A protein with 75% or more homology in amino acid sequence and the same function. The present invention discovers a new gene PwNAC2, and introduces it into Arabidopsis thaliana to obtain PwNAC2-transformed Arabidopsis plants. Experiments prove that the drought tolerance and salt tolerance of PwNAC2-transferred Arabidopsis plants are significantly improved, indicating that PwNAC2 or its encoded protein With stress resistance.
Description
technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a transcription factor PwNAC2 related to plant stress tolerance, its coding gene and its application. Background technique [0002] Abiotic stresses such as drought and salinity seriously affect the normal growth and development of plants, and plants will develop a series of effective mechanisms during the long-term evolution process, including regulating the expression of stress-related genes to cope with external adverse environments. Stress-induced expression genes can be roughly divided into two categories: functional protein-coding genes and transcriptional regulatory protein-coding genes involved in stress signal transduction. [0003] NAC transcription factor is a plant-specific transcriptional regulator with multiple biological functions, and plays an important role in plant growth and development, morphogenesis, hormone regulation and stress response. At present,...
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