转移巴西橡胶树HbCBF1基因提高烟草抗寒能力的研究

通过农杆菌介导的叶盘转化法将巴西橡胶树HbCBF1基因导入到烟草中增强表达。GUS染色和PCR鉴定结果表明,外源DNA片段已成功导入并整合到烟草基因组中。选取3个烟草转基因株系用于进一步鉴定分析,分析表明,转化植株游离脯氨酸含量与对照相比,分别上升了4.2倍、3.1倍和2.5倍,叶绿素A和叶绿素B含量也明显提高。电导率法测定转化烟草的相对存活率分别为19.8%、17.5%、16.3%,高于对照植株的7.7%。以上结果表明,橡胶树HbCBF1基因能够增强非低温驯化植物烟草的抗寒能力,其机理可能是通过促进细胞内容物的积累从而提高细胞的低温防御能力。

Transfer of the Hevea brasiliensis HbCBF1 Gene Enhanced the Cold Tolerance in Tobacco

Rubber tree(Hevea brasiliensis Muell. Arg.)HbCBF1 gene was transferred into tobacco through Agrobacterium mediated leaf disc transformation method. GUS staining and PCR amplification results indicated that the exogenous DNA fragment was transferred and integrated into the tobacco genome. Three transgenic lines were selected for further analysis. The results showed that the free proline content was increased 4.2, 3.1 and 2.5 folds respectively, and the chlorophyll A and B content also increased significantly. Electrolyte leakage analysis demonstrated the transgenic plants had relative survival rates of 19.8%, 17.5% and 16.3% respectively, which was far higher than that of control. All these experiments results proved that rubber tree HbCBF1 gene could enhanced the cold tolerance of the non-acclimate tobacco by possibly promoting the accumulation of metabolites content.