State Key Laboratory of Genetic Engineering, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai 200433, China.
J Plant Physiol. 2012 Sep 15;169(14):1408-16. doi: 10.1016/j.jplph.2012.05.016. Epub 2012 Jul 12.
Low temperature is among the most significant abiotic stresses, restricting the habitats of sessile plants and reducing crop productivity. Cold regulated (COR) genes are low temperature-responsive genes expressing under regulation of a specific signal transduction pathway, which is designated C-repeat-binding-factor (CBF) signaling pathway. In the present article, cold bioassay showed that the transcript level of cold regulated gene CbCOR15b from shepherd's purse (Capsella bursa-pastoris) was obviously elevated under cold treatments. Reverse transcription-PCR (RT-PCR) and GUS report system revealed that unlike AtCOR15b, CbCOR15b expressed not only in leaves but also in stems and maturation zone of roots. When transgenic tobacco plants ectopically expressing CbCOR15b were exposed to chilling and freezing temperatures, they displayed more cold tolerance compared to control plants. According to the electrolyte leakage, the relative water content, the glucose content and the phenotype observation, CbCOR15b transformants suffered less damage under cold stress. Further investigation of the subcellular localization of CbCOR15b by transient expression of fusion protein CbCOR15b-GFP revealed that it was localized exclusively in the chloroplasts of tobacco mesophyll cells and in the cytoplasm of onion epidermal cells. It can be concluded that CbCOR15b which located in the chloroplasts and in the cytoplasm of cells without chloroplasts was involved in cold response of C. bursa-pastoris and conferred enhanced cold tolerance in transgenic tobacco plants.
低温是最重要的非生物胁迫之一,限制了固着植物的栖息地并降低了作物的生产力。冷调节(COR)基因是低温响应基因,在特定信号转导途径的调控下表达,该途径被指定为 C-重复结合因子(CBF)信号途径。在本文中,冷生物测定表明,来自荠菜(Capsella bursa-pastoris)的冷调节基因 CbCOR15b 的转录水平在低温处理下明显升高。反转录 - PCR(RT-PCR)和 GUS 报告系统显示,与 AtCOR15b 不同,CbCOR15b 不仅在叶片中表达,而且在茎和根成熟区中表达。当异位表达 CbCOR15b 的转基因烟草植物暴露于冷胁迫和冷冻温度时,与对照植物相比,它们表现出更高的耐寒性。根据电解质泄漏、相对水含量、葡萄糖含量和表型观察,CbCOR15b 转化体在冷胁迫下受到的损伤较小。通过瞬时表达融合蛋白 CbCOR15b-GFP 对 CbCOR15b 的亚细胞定位的进一步研究表明,它仅定位于烟草叶肉细胞的叶绿体和洋葱表皮细胞的细胞质中。可以得出结论,定位于叶绿体和无叶绿体细胞的细胞质中的 CbCOR15b 参与了荠菜的冷响应,并赋予了转基因烟草植物增强的耐寒性。