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ROP(Rho of plant)作为植物中唯一具有信号传递功能的小GTP结合蛋白,在极性生长、发育和环境应答等过程中具有重要作用.本文采用酵母双杂交技术,从水稻幼穗筛选出2种与水稻ROP家族成员OsRac5互作蛋白的基因OsMY1和OsMY2.序列分析显示,OsMY1与OsMY2的核苷酸、氨基酸序列的相似性分别为14%和49%,均含有卷曲螺旋结构域;GST pull down分析显示,OsMY1可以和激活型OsRac5在体外结合;实时定量PCR分析显示,OsMY1和OsMY2在水稻生长发育时期的根、茎、叶中广泛表达,尤其在幼穗中的表达量显著高于其它组织.干旱、低温、高盐等非生物胁迫和植物激素吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA)处理能不同程度地上调OsMY1和OsMY2基因在水稻幼苗中的表达,但油菜素内酯(BR)、水杨酸(SA)、6-苄氨基嘌呤(6-BA)对2种基因的表达没有显著的影响.本文为进一步研究OsMY1和OsMY2基因在水稻生长发育、植物激素和胁迫应答中的功能提供了重要依据,为研究OsMY1、OsMY2和OsRac5蛋白质相互作用的功能联系和作用机制奠定了基础.
As the only small GTP-binding protein with signal transduction in plants, ROP (Rho of plant) plays an important role in the process of polar growth, development and environmental response.In this paper, yeast two-hybrid technique was used to screen 2 OsMY1 and OsMY2, which interact with OsRac5, a member of rice ROP family, showed that the nucleotide and amino acid sequence identities of OsMY1 and OsMY2 were 14% and 49%, respectively, and all contained coiled-coil domains. GST pull down analysis showed that OsMY1 could bind with activated OsRac5 in vitro. Real-time quantitative PCR analysis showed that OsMY1 and OsMY2 were widely expressed in roots, stems and leaves during rice growth and development, especially in panicles. The abiotic stress such as drought, low temperature and high salt, and the IAA, ABA and GA treatments up-regulated the expression of OsMY1 and OsMY2 in rice seedlings (BR), salicylic acid (SA), 6-benzylaminopurine (6-BA) had no significant effect on the expression of these two genes.In order to further study the effects of OsMY1 and OsMY2 on rice growth and development ,plant Hormone and stress response functions provides an important basis for the study OsMY1, functional linkages and mechanisms of protein-protein interactions OsMY2 and OsRac5 basis.