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为研究冷诱导基因的转录因子CBFI(C-repeat binding factor)基因对植物抗寒的作用,利用PCR技术从拟南芥菜(Arabidopsis thaliana)中扩增并克隆了该转录因子,并将其与CaMV 35S启动子融合构建成植物表达载体pBPCBF1。以农杆菌介导的叶盘法,分别转化了油菜和烟草。经PCR程序的DNA分析和Southern杂交对具有卡那霉素抗性的再生植株进行了鉴定,表明CBFl基因已整合进烟草和油菜基因组中。以电解质渗漏法分别检测了转化的油菜和烟草的抗寒性,结果显示转基因油菜的抗寒性较未转基因油菜有明显提高,转基因烟草抗寒性也有一定的提高。因此利用对转录因子的调控来提高植物的抗寒性可能有很好的应用前景。
In order to study the role of the C-repeat binding factor (CBFI) cold-inducible gene in plant cold tolerance, the transcription factor was amplified from Arabidopsis thaliana by PCR and cloned into CaMV 35S promoter fusion construct into plant expression vector pBPCBF1. Agrobacterium-mediated leaf disc method, respectively, transformed into rape and tobacco. The kanamycin-resistant regenerated plants were identified by DNA analysis of the PCR program and Southern blotting, indicating that the CBF1 gene has been integrated into the genome of tobacco and rape. Cold tolerance of transformed rapeseed and tobacco was tested by electrolyte leakage method. The results showed that the cold resistance of transgenic rapeseed was significantly higher than that of non-transgenic rapeseed, and the cold resistance of transgenic tobacco was also improved. Therefore, the use of transcription factor regulation to improve the plant’s cold resistance may have good application prospects.