Drought resistance-associated protein, encoding gene thereof and application of drought resistance-associated protein and encoding gene thereof in regulation of plant drought resistance
A technology related to proteins and transgenic plants, applied in applications, plant peptides, plant products, etc., can solve the problems of complexity of drought tolerance traits, inability to indicate functional genes, deepening and other problems
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Embodiment 1
[0156] Embodiment 1, the construction of transgenic Arabidopsis thaliana
[0157] This embodiment provides a drought-resistance-related protein gene derived from wild-type Colombian ecotype Arabidopsis thaliana (named col), the gene name is CHYR1 gene, and the genomic DNA sequence of the CHYR1 gene is shown in sequence 1 in the sequence table, the sequence 1 consists of 2275 nucleotides, of which the 230-325th, 414-540th, 601-685th, 719-803rd, 882-958th, 1003-1107th, 1st Positions 1149-1234, 1318-1401, 1494-1580, 1651-1730 and 1773-1873 are the sequences of 11 introns of the CHYR1 gene, respectively. The cDNA sequence of the CHYR1 gene is shown in sequence 2 in the sequence listing, and sequence 2 consists of 1262 nucleotides, wherein the 153-1028th position of sequence 2 is the drought resistance-related protein (ie CHYR1) shown in sequence 3 in the coding sequence listing , sequence 3 consists of 291 amino acid residues.
[0158] 1. Construction of recombinant vector and r...
Embodiment 2
[0171] Example 2, CHYR1 can improve the drought resistance of Arabidopsis
[0172] The effects of CHYR1 gene on the drought resistance of Arabidopsis thaliana were detected by transgenic Arabidopsis thaliana and CHYR1 gene-deleted Arabidopsis mutants. The transgenic Arabidopsis thaliana used in this embodiment is the T 2 Generation OE35 and T 2 Substitute OE42, and place T 2 Generation VC was used as a control; the CHYR1 gene-deleted Arabidopsis mutants used in this example were two T-DNA insertion mutants with CHYR1 gene expression loss, and their names were chyr1-1 and chyr1-2, chyr1-1 The genetic background of and chyr1-2 is the wild type Arabidopsis thaliana (named col), and col is used as a control.
[0173] 1. Drought tolerance experiment
[0174] Fully mix the vermiculite and nutrient soil according to the ratio of 1:1, weigh 250g and put them into each small plate (12cm×12cm, 6 in total) of the black seedling tray with a flat bottom, put it into a large tray and fi...
Embodiment 3
[0197] Example 3, CHYR1 T178D Can improve the drought resistance of Arabidopsis
[0198] The protein obtained by mutating threonine at position 178 of sequence 3 into alanine is named CHYR1 T178A , the DNA molecule encoding the protein is a DNA molecule obtained by mutating the 684-686th nucleotide (i.e. act) in the 153-1028th position of sequence 2 into gct, and the DNA molecule is named CHYR1 T178A Gene; the protein obtained by mutating the 178th threonine in sequence 3 to aspartic acid is named CHYR1 T178D , CHYR1 T178D The sequence of the protein is shown in sequence 5, the DNA molecule encoding the protein is the DNA molecule obtained by mutating the 684-686th nucleotide (i.e. act) in the 153-1028th position of sequence 2 into gat, the DNA molecule named CHYR1 T178D gene, CHYR1 T178D The sequence of the gene is shown in SEQ ID NO:4.
[0199] 1. Construction of transgenic plants
[0200] Replace the fragment between the Bam HI and Xho I recognition sequences of the ...
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