水稻多胺转运蛋白OsPUT1基因RNAi载体的构建及遗传转化

Construction and Genetic Transformation of RNAi Vector for OsPUT1 Gene in Rice

在植物中,多胺(腐胺,亚精胺和精胺)参与许多生理过程,包括胚胎发育、器官形成、花芽发育,叶片衰老,花粉管生长、果实的发育和成熟,生物和非生物胁迫的响应。但是,PAS具有内在毒性和多重功能,细胞内的PA含量受到严格控制,受转运体的调节。因此,在水稻中研究水稻多胺转运蛋白OsPUT1的功能至关重要。本研究以水稻多胺转运蛋白基因OsPUT1为研究对象,通过构建OsPUT1-RNAi载体,借助根癌农杆菌介导水稻遗传转化,将OsPUT1的RNAi表达框转到水稻基因组中;经过PPT抗性筛选与PCR鉴定,成功获得了34株转基因水稻。对其中的7株转基因苗进一步作RT-PCR检测,显示有5株OsPUT1基因表达下调,实现了设计目标。这些转基因水稻为进一步研究OsPUT1的功能与作用机制提供了良好的遗传材料。

The polyamines(putrescine, spermidine and spermine) involves in many physiological processes in plants,including embryonic development, organ formation, development of flower bud, leaf senescence, pollen tube growth, fruit development and maturation, biotic and abiotic stress response. However, because of the intrinsic toxicity and multiple functions of PAS, the content of PA in cells is strictly controlled and regulated by transporter.Therefore, it is essential to study the function of OsPUT1 in rice. In this research, OsPUT1 was used as material,the RNAi expression box of OsPUT1 was transferred to rice genome via constructing of RNAi plasmid and with the help of agrobacterium-mediated transformations. Through resistance selection of PPT and PCR identification,thirty-four transgenic plants were gained. The RT-PCR results of 7 plants selected from the transgenic plants showed that the OsPUT1 was significantly inhibited in 5 strains, which suggested the design objectives were achieved.These transgenic plants were the good materials for further research on function and mechanism of OsPUT1.