Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, 630-0192 Japan.
Plant Cell Physiol. 2010 Apr;51(4):585-95. doi: 10.1093/pcp/pcq024. Epub 2010 Mar 4.
Plant-specific Rac/Rop small GTPases function as molecular switches for numerous signal transduction events, including defense responses. To understand the function of each of the seven Rac/Rop family members in rice, we studied the tissue-specific expression patterns of Rac/Rop genes by semi-quantitative reverse transcription-PCR (RT-PCR), and also Rac/Rop subcellular localization using green fluorescent protein (GFP) fusion proteins in transient expression systems. We also investigated the roles of these genes in disease resistance by testing single Rac/Rop-RNAi (RNA interference) plants against the rice blast pathogen Magnaporthe grisea. Our studies show that expression of OsRac2, 6 and 7 is very low in leaf blades, and reveal a strong correlation between the number of lysine and/or arginine (KR) residues in the polybasic region of Rac/Rop GTPases and their subcellular distribution in vivo. Infection assays showed that OsRac1 is a positive regulator of blast resistance, confirming previous observations, whereas OsRac4 and OsRac5 are negative regulators of blast resistance. OsRac6 may make minor contributions to disease resistance, while OsRac3 and OsRac7 are probably not involved in defense. Therefore, our study suggests that the rice Rac/Rop family plays multiple roles in diverse cellular activities and has both positive and negative functions in disease resistance.
植物特异性 Rac/Rop 小 GTPases 作为众多信号转导事件的分子开关,包括防御反应。为了了解水稻中 7 个 Rac/Rop 家族成员中的每一个的功能,我们通过半定量 RT-PCR(反转录 PCR)研究了 Rac/Rop 基因的组织特异性表达模式,并用绿色荧光蛋白(GFP)融合蛋白在瞬时表达系统中研究了 Rac/Rop 的亚细胞定位。我们还通过测试单个 Rac/Rop-RNAi(RNA 干扰)植物对稻瘟病菌 Magnaporthe grisea 的抗性来研究这些基因在疾病抗性中的作用。我们的研究表明,OsRac2、6 和 7 在叶片中的表达水平很低,并且 Rac/Rop GTPases 中的碱性氨基酸(KR)残基数与它们在体内的亚细胞分布之间存在很强的相关性。感染实验表明,OsRac1 是稻瘟病抗性的正调控因子,这与先前的观察结果一致,而 OsRac4 和 OsRac5 是稻瘟病抗性的负调控因子。OsRac6 可能对疾病抗性有较小的贡献,而 OsRac3 和 OsRac7 可能不参与防御。因此,我们的研究表明,水稻 Rac/Rop 家族在多种细胞活动中发挥多种作用,并在疾病抗性中具有正、负功能。