| Genetic transformation of wheat with dehydrin wzy1-2 gene and the expression level at different growth stages under drought stress |
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| DOI：10.7606/j.issn.1000-7601.2019.06.01 |
| Key Words: dehydrin wheat water stress particle bombardment drought\|inducible gene |
| Author Name | Affiliation | | TIAN Ye | College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Area, Northwest A & F University, Yangling, Shaanxi 712100, China | | YU Zhengyang | College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Area, Northwest A & F University, Yangling, Shaanxi 712100, China | | SHI Xueying | College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Area, Northwest A & F University, Yangling, Shaanxi 712100, China | | QI Yuhong | College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Area, Northwest A & F University, Yangling, Shaanxi 712100, China | | ZHANG Dapeng | College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Area, Northwest A & F University, Yangling, Shaanxi 712100, China | | ZHANG Linsheng | College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Area, Northwest A & F University, Yangling, Shaanxi 712100, China |
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| Abstract: |
| Based on the principle of RNA interference and the homology analysis, a 298 bp cDNA sequence between 0~298 bp in the dehydrin wzy1-2 gene was selected as the interference fragment, and then cloned interfering fragment was inserted into the efficient plant expression vector pTCK303, clone site, to successfully constructed RNA interference expression vectors containing inverted repeats. Using immature embryos of Zhengyin 1 wheat as explants, 2512 immature embryo calli was bombarded by particle bombardment, and 26 regenerated plants were obtained with a regeneration rate of 1.03%. Using the specific primers of the Hpt gene on the expression vector, 26 T0 generation regenerated plants were subjected to PCR analysis. Six Hpt gene positive plants were detected, and the positive conversion rate of the Hpt gene was 0.24%. Through quantitative real\|time analysis of the dehydrin wzy1-2 gene, it was found that the expression of the target gene significantly decreased. To lay the foundation for further study of dehydrin wzy1-2 gene function, two drought tolerant wheat varieties(Shaanhe 6 and Zhengyin 1), were treated with drought stress at four growth stages (seedling, tillering, jointing, and flowering), and expression of wheat WZY1-2 protein was studied. The expression level of WZY1-2 protein in wheat incread with increasing drought degree and at the seedling stage it was higher than that in the other three growth stages. Western blot results showed that the size of the non\|specific bands of Shaanhe 6 and Zhengyin 1 were different. The wheat wzy1-2 derived from the molecular level was a drought\|inducible gene. |
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