拟南芥AtCOR15a转化烟草及后代抗寒性研究

【目的】研究拟南芥AtCOR15a基因的抗寒功能。【方法】构建pCAMBIA1301-COR15a的植物表达载体,利用农杆菌介导的叶盘法转化烟草,通过抗生素筛选和PCR检测获得转基因烟草。低温胁迫后,利用蒽酮法和酸式茚三酮法测定转基因烟草后代与野生型烟草的可溶性糖和脯氨酸含量,同时观察其抗寒能力。【结果】转基因烟草后代可溶性糖含量和脯氨酸含量均比野生型烟草有所提高,并在第5 d达到峰值。低温处理后转基因烟草后代能正常生长,而野生型烟草则全部死亡。【结论】生理生化指标的测定发现,可溶性糖含量与脯氨酸含量的变化趋势与烟草的抗寒性呈正相关,AtCOR15a基因导入烟草中可以明显提高烟草对低温胁迫的抵抗力。

Study on AtCOR15α Gene from Arabidopsis in Tobacco and the Cold Tolerance of Its Offspring

【Objective】In order to study the cold tolerance function of AtCOR15a from Arabidopsis.【Method】The plant expression vector of pCAMBIA1301-COR15a was constructed,and then it was transferred into tobacco genome through agrobacterium tumefaciens.Antibiotic selection and PCR detection was used for detecting transgenic tobacco.It was to explore the effect of low temperature stress on soluble sugar and proline content of transgenic and wild-type tobacco by determination of anthrone method and the acid ninhydrin,and observe their cold tolerance at the same time.【Result】It was found that soluble sugar content and proline content of transgenic tobacco were higher than those of wild-type tobacco,and reached peak value at day 5.After 4℃and-1℃stress,the transgenic tobacco plants can grow normally,while all wild-type tobacco plants died.【Conclusion】The determination of the physiological and biochemical data show that the changing trend of soluble sugar content and proline content were positively correlated to cold tolerance of tobacco,and the results suggested that expression of AtCOR15a gene significantly increased the cold tolerance of transgenic tobacco.