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

CN111826390AActive Publication Date: 2020-10-27INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2020-10-27

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Abstract

The invention discloses application of protein WRKY78 to regulation of the biological stress resistance of plants. An amino acid sequence of the protein WRKY78 is as shown in sequence 2 in a sequencetable. The expression quantity and / or activity of the protein WRKY78 in oryza sativa L. spp. japonica are improved to obtain transgenic rice A; compared with the oryza sativa L. spp. japonica, the transgenic rice A is significantly improved both in rice stripe disease resistance and rice blast resistance; the expression quantity and / or activity of the protein WRKY78 in the oryza sativa L. spp. japonica are inhibited to obtain transgenic rice B; and compared with the oryza sativa L. spp. japonica, the transgenic rice B is significantly reduced in the aspects of rice stripe disease resistance, rice blast resistance and laodelphax striatellus resistance. Thus it can be seen that the protein WRKY78 can regulate the biological stress resistance of the plants. The protein WRKY78 has important application value.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to the application of protein WRKY78 in regulating plant biological stress resistance. Background technique

[0002] Rice (Oryza sativa L.) is one of the most important food crops in the world and is the staple food for more than half of the world's population. In the breeding process of human beings for thousands of years, high yield, stable yield and excellent yield have always been the three important goals of rice variety breeding, and they are also the main goals pursued by agricultural production. In the process of rice production, due to the single planting method, rice is easily affected by biotic stress, which seriously affects the yield and quality of rice.

[0003] Biological stress refers to the general term of various biological factors that are unfavorable to plant survival and development, usually caused by infection and competition, such as diseases, insect ...

Examples

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. ...