摘要
目的:为了探讨水稻生长素极性运输输出载体蛋白OsPIN1a的在水稻不同组织的作用和分布。方法:以带有GFP和GUS标记的转OsPIN1a基因水稻和野生型中花11水稻为研究材料。通过PCR方法检测抗性标记基因潮霉素基因判断植株是否阳性;通过半定量RT-PCR方法分析OsPIN1a基因在转基因植株幼穗、叶片、根系的表达;通过GUS组织染色检测OsPIN1a在幼穗、叶片、根系中的活性;通过激光共聚焦显微镜观察OsPIN1a-GFP蛋白在根尖亚细胞定位情况。结果:实验所用的水稻株系均为转基因株系;RT-PCR检测发现,在根尖、叶片和幼穗中hpt和OsPIN1a基因均有表达;GUS组织染色结果表明,幼穗、叶片、根系均有GUS活性,其中在幼嫩的组织中表达量最大,如刚萌发种子胚芽鞘、花药和柱头;GUS活性受外源生长素诱导而受生长素极性运输抑制剂(TIBA)的抑制;激光共聚焦显微镜观察发现,OsPIN1a-GFP蛋白主要分布在根尖细胞膜上。结论:实验表明OsPIN1a基因参与了水稻各个器官和组织的发育,这些组织发育可能都受到生长素极性运输的调控。
Objective：In order to explore the function and distribution of auxin efflux carrier protein OsPIN1a in different tissues of rice. Method：The Japonica rice Zhonghua 11 and its over -expressed OsPIN1a gene strains with GFP and GUS reporter gene were chosen as materials. PCR method was used to detect the hygromycin resistance gene marker for transgenetic plants, semi - quantitative RT - PCR was used to detect the expression level of OsPIN1 a in rice root,leaf and panicle, the GUS activity in rice root, leaf and panicle was detected by using GUS staining, and subcellular localization of OsPIN1 a - GFP in the root tip observed by a laser confocal - microscopy~ Result：The results showed that all of transgenic strains were positive by genomic PCR detection; the hygromycin resistance gene marker and OsPIN1 a gene both expressed in rice roots,leaves and panicles by semi - quantitative RT - PCR. By GUS activity staining we found that panicles, leaves and roots all displayed GUS activity. Besides ,the expression of GUS were stronger in young tissues, such as the newly -germinated seeds,coleoptile and stigma. The GUS activity were also induced by IAA and suppressed by polar auxin transport inhibitors （TIBA） . Moreover, the OsPIN1 a - GFP protein mainly located on the cell membrane in rice root tips by a laser confocal - microscopy. Conclusion： The experimental results showed that OsPIN1a gene might involve in the development and morphogenesis of these tissues by polar auxin transport.
出处
《生物技术》
CAS
CSCD
北大核心
2014年第1期27-32,共6页
Biotechnology
基金
国家自然科学基金项目( "生长素极性运输与钙信号对水稻根负向光性影响"
No. 31071353)
绍兴文理学院科研启动经费( "OsPIN1a基因在水稻根生长 及 负 向 光 性 中 的 作 用 研 究 "
No. 20135004) 资助~~
