烟草soc1基因的克隆和表达分析.pdfVIP

石永春等　烟草SOC1 基因的克隆和表达分析 99 生物技术 烟草SOC 1 基因的克隆和表达分析 ， ， 石永春 杨永银 刘卫群 河南农业大学生命科学学院，郑州文化路95 号450002 摘　要：为明确SOC1 基因在叶片中的生理作用，克隆到烟草中SOC1 基因的完整开放阅读框，全长806 bp 。农杆菌介导转化烟 草后，获得26 棵转基因植株，其中23 棵鉴定为阳性植株，转化率为88.46% 。与野生型烟草K326 相比，获得的SOC1 过表达烟 株开花提前，株高增加，叶片宽大。半定量RT-PCR 结果显示，SOC1 过表达不影响叶片中Rubisco 大亚基、蔗糖磷酸合成酶和 蔗糖合成酶基因的表达水平，但提高了抗坏血酸氧化酶的表达水平；提高了碳酸酐酶活性、蔗糖含量和还原性抗坏血酸含量，并 降低了抗坏血酸过氧化物酶活性。表明SOC1 可能通过影响氧化还原状态而提高碳酸酐酶活性和光合速率。 关键词：烟草；SOC1 ；转基因；光合；抗坏血酸 doi:10.3969/j .issn.1004-5708.2014.02.017 中图分类号：Q78 ；S572.01　　　文献标志码：A　　　文章编号：1004-5708(2014)02-0099-05 Cloning and expression of SOC1 gene in tobacco SHI Y ongchun, YAN G Y ongyin, LIU W eiqun College of Life Science, Henan Agricultural Univer sity ，Zhengzhou 450002, China Abstract: Open reading frame of SUPPRESSOR OF OVEREXPRESSION OF CON STANTS 1 (SOC 1) gene in tobacco was cloned with full length of 806 bp to investigate its physiological function in tobacco. Agrobacterium-mediated leaf-disk transformations were used to create 26 SOC1 transgenic tobacco plants. Among which 23 were identified as positive plants with transformation rate of 88.46%. Compared with tobacco variety K326, SOC1 over-expression plants had early floral time, higher plant heights and wider and longer leaves. Semi-quantitative RT-PCR results showed that over-expression of SOC1 did not affect expression levels of Rubisco large subunit, sucrose phosphate synthetase and sucrose synthetase in leaves, but increased the level of ascorbare oxidase. It also increased carbonic anhydrase activity, concentrations of sucrose and ascorbic acid, and reduced ascorbate peroxidase activities. These results suggested that SOC 1 increase