National Agri-Food Biotechnology Institute, Mohali 140306, Punjab, India; Department of Biotechnology, Panjab University, Chandigarh, India.
National Agri-Food Biotechnology Institute, Mohali 140306, Punjab, India.
Plant Sci. 2022 Nov;324:111413. doi: 10.1016/j.plantsci.2022.111413. Epub 2022 Aug 10.
The rice Hybrid Proline Rich Protein (HyPRP) encoding gene, OsHyPRP16 expression exhibit early upregulation in response to Magnaporthe oryzae inoculation. Here, we functionally characterized the OsHyPRP16 promoter through deletion analysis in transgenic Arabidopsis using GUS (β-glucuronidase) reporter assay. The promoter fragments, sequentially deleted from the 5' end could induce differential GUS activity in response to stresses induced by different hormones and abiotic stress conditions. In addition, a strong GUS induction was observed in M. oryzae inoculated transgenic Arabidopsis. Based on the insilico and stress-inducibility of D1 promoter fragment against various phytohormones and rice blast fungus, and with no basal activity under control conditions, we rationally selected D1 promoter fragment to drive the expression of a major rice blast resistance gene; Pi54 in the genetic background of blast susceptible TP309 rice line. The D1 promoter fragment was able to induce the expression of Pi54 at immediate-early stages of M. oryzae infection in transgenic rice. The transgenic plants with Pi54 under the control of D1 promoter fragment displayed complete resistance against M. oryzae infection as compared to control plants. The present study suggests that the D1 fragment of OsHyPRP16 promoter is a valuable tool for breeding and development of rice lines with early-inducible and pathogen-responsive enhanced disease resistance.
水稻杂种脯氨酸丰富蛋白(HyPRP)编码基因 OsHyPRP16 的表达在受到稻瘟病菌接种后早期上调。在这里,我们通过在转基因拟南芥中使用 GUS(β-葡萄糖醛酸酶)报告基因分析进行了 OsHyPRP16 启动子的功能表征。通过从 5'端依次缺失启动子片段,可以诱导对不同激素和非生物胁迫条件诱导的胁迫的差异 GUS 活性。此外,在接种稻瘟病菌的转基因拟南芥中观察到强烈的 GUS 诱导。基于 D1 启动子片段对各种植物激素和稻瘟病菌的顺式作用元件和应激诱导性,以及在对照条件下没有基础活性,我们合理选择 D1 启动子片段来驱动主要稻瘟病抗性基因 Pi54 在感病 TP309 水稻品系中的表达。D1 启动子片段能够在 M. oryzae 感染的早期诱导 Pi54 在转基因水稻中的表达。与对照植物相比,受 D1 启动子片段控制的 Pi54 的转基因植物对 M. oryzae 感染表现出完全抗性。本研究表明,OsHyPRP16 启动子的 D1 片段是培育和开发具有早期诱导和病原体响应增强抗病性的水稻品系的有价值的工具。