论文部分内容阅读
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: To investigate the role and distribution of OsPIN1a, a polar exporter of auxin in rice, in different tissues of rice. Methods: OsPIN1a transgenic rice with GFP and GUS markers and wild type Zhonghua 11 were used as materials. The expression of OsPIN1a gene in panicles, leaves and roots of transgenic plants was analyzed by semi-quantitative RT-PCR. The expression of OsPIN1a was detected in young spikes and leaves by GUS staining , Root activity; observed by laser confocal microscopy of OsPIN1a-GFP protein in the apical subcellular localization. Results: All of the rice lines used in the experiment were transgenic lines. The expression of hpt and OsPIN1a genes in root tips, leaves and young ears were detected by RT-PCR. The results of GUS staining showed that young panicles, GUS activity was highest in young tissues, such as germ sheath, anther and stigma; GUS activity was induced by exogenous auxin and inhibited by TIBA; laser Confocal microscopy showed that OsPIN1a-GFP mainly distributed on the apical membrane. Conclusion: The experiment indicated that OsPIN1a gene is involved in the development of various organs and tissues of rice. The development of these tissues may all be regulated by auxin polar transport.