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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。
In plants, polyamines (putrescine, spermidine and spermine) are involved in many physiological processes including embryonic development, organogenesis, flower bud development, leaf senescence, pollen tube growth, fruit development and maturation, biotic and abiotic stresses the response to. However, PAS is inherently toxic and has multiple functions, and intracellular PA levels are tightly controlled and regulated by transporters. Therefore, it is important to study the function of rice polyamine transporter OsPUT1 in rice. In this study, OsPUT1, a gene of rice polyamine transporter, was used as the research object. The OsPUT1-RNAi vector was constructed and Agrobacterium tumefaciens was used to mediate rice genetic transformation. OsPUT1 RNAi expression cassette was transferred into rice genome. PCR identification, successfully obtained 34 transgenic rice. Seven of the transgenic plants were further subjected to RT-PCR. The results showed that the expression of five OsPUT1 genes was down-regulated and the design goal was achieved. These transgenic rice provide a good genetic material for further studying the function and mechanism of OsPUT1.