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为探究香樟Cc CBFb基因增强植株非生物胁迫抗性的功能,通过农杆菌介导法将该基因转入烟草中,经PCR和半定量RT-PCR技术鉴定阳性转基因株系后,对获得的T1代转基因无性系(T2和T4株系)以及野生型(WT)进行干旱(0、150、300和450 mmol·L~(-1)的甘露醇)、高盐(0、100、200和300 mmol·L~(-1)的Na Cl)、4℃低温、–4℃冰冻胁迫处理,结果显示:转基因烟草在干旱和高盐胁迫下,幼苗的存活率均高于野生型;经4℃低温处理后,转基因烟草植株的脯氨酸和可溶性糖含量均显著高于野生型植株,而丙二醛(MDA)含量均低于野生型植株;经–4℃的冰冻处理6 h后,野生型和转基因T4株系植株叶片均出现不同程度的萎蔫,而转基因T2株系植株未出现不良现象。由此可见,过表达CcCBFb基因不但能够增强烟草的抗旱和抗盐性,而且能够显著增强抗寒性。
In order to explore the function of Cc CBFb gene of C. camphora in enhancing plant abiotic stress resistance, the gene was transferred into tobacco by Agrobacterium tumefaciens-mediated method. After the positive transgenic lines were identified by PCR and semi-quantitative RT-PCR, T1 generation transgenic clones (T2 and T4 lines) and wild type (WT) were subjected to drought (0, 150, 300 and 450 mmol·L -1 mannitol), high salt (0, 100, 200 and 300 mmol·L -1 NaCl), 4 ℃ low temperature and -4 ℃ freezing stress. The results showed that under drought and salt stress, the survival rate of transgenic tobacco was higher than that of wild type. After 4 ℃, the contents of proline and soluble sugar in the transgenic tobacco plants were significantly higher than those in the wild type plants, while the content of malondialdehyde (MDA) in the transgenic tobacco plants was lower than that in the wild type plants. After being frozen at -4 ℃ for 6 h, The leaves of wild-type and transgenic T4 plants showed varying degrees of wilting, while the transgenic T2 plants showed no adverse effects. Thus, overexpression of CcCBFb gene can not only enhance tobacco drought and salt resistance, but also significantly enhance the cold resistance.