Application of protein WRKY78 to regulation of biological stress resistance of plants
A biological stress and protein technology, applied in angiosperms/flowering plants, applications, plant products, etc., can solve problems such as environmental pollution, increased rice production costs, and rapid variation of physiological races
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Embodiment 1
[0082] Embodiment 1, rice stripe blight resistance, rice blast resistance and SBPH resistance improvement of transgenic WRKY78 rice
[0083] 1. Construction of recombinant plasmid 35S:WRKY78-YFP
[0084] 1. Artificially synthesize the DNA double-stranded molecule shown in sequence 1 in the sequence listing and use it as a template, using a primer pair consisting of upstream primer F: 5'-CAAGGGATCCATGGCCGATTCGCCAAACCCTAGCT-3' and downstream primer R: 5'-CAAGGCGGCCGCCACGGACCCATGACTAAAT-3' Perform PCR amplification to obtain PCR amplification products.
[0085] 2. After step 1 is completed, the PCR amplification product is taken, and a DNA fragment of about 1880 bp is recovered.
[0086] 3. Ligate the DNA fragment recovered in step 2 and the pENTR-3C cloning vector by T4 DNA ligase to obtain an intermediate vector.
[0087] 4. After completing step 3, perform LR reaction with the intermediate vector and the pH7-WG2Y vector to obtain the recombinant plasmid 35S:WRKY78-YFP.
[008...
Embodiment 2
[0126] Example 2. Rice stripe blight resistance, rice blast resistance and BPH resistance of mutants knocking down the WRKY78 gene in rice are reduced
[0127]wrky78#6 and wrky78#8 are rice knockdown mutants of WRKY78 gene obtained from selfing first generation of OsWRKY78-RNAi plants. OsWRKY78-RNAi plants are described in the following literature: Chang-Quan Zhang, Yong Xu, YanLu, Heng-Xiu Yu, Ming-Hong Gu, Qiao-QuanLiu. The WRKY transcription factor OsWRKY78 regulates stem elongation and seed development in rice. Planta 234:541 -554., named "OsWRKY78-RNAi plants" in the literature. Both wrky78#6 and wrky78#8 were provided by Professor Liu Qiaoquan of Yangzhou University.
[0128] 1. Real-time fluorescent quantitative PCR detection of relative expression of WRKY78 gene
[0129] The rice seeds to be tested are the seeds of wrky78#6, the seeds of wrky78#8 or the seeds of Nipponbare.
[0130] 1. Same as 1 in Step 3 of Example 1.
[0131] 2. Same as 2 in Step 3 of Example 1. ...
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