向日葵转录因子HB-12的定位分析及过表达验证  

Subcellular Localization Analysis and Overexpression Verification of Transcription Factor HB-12 from Sunflower (Helianthus annuus L.)

作　　者：孙瑞芬[1] 张艳芳[1] 郭树春[1] 李素萍[1] 于海峰[1] 聂惠[1] 牛素清[1] 乔慧蕾[1] 安玉麟[1] Sun Ruifen;Zhang Yanfang;Guo Shuchun;Li Suping;Yu Haifeng;Nie Hui;Niu Suqing;Qiao Huilei;An Yulin(Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences,Huhhot 010031,China)

机构地区：[1]内蒙古农牧业科学院,呼和浩特010031

出　　处：《中国细胞生物学学报》2018年第9期1526-1536,共11页Chinese Journal of Cell Biology

基　　金：内蒙古农牧业科技创新基金(批准号:2013CXJJN14)资助的课题~~

摘　　要：向日葵转录因子HB-12属于HD-Zip I类转录因子家族,在植物逆境胁迫应答中发挥着极其重要的作用。该研究构建HB-12基因瞬时表达载体并进行亚细胞定位分析,表明HB-12基因定位于细胞核。构建HB-12基因植物超表达载体转化野生型烟草获得了转基因植株。进行了Na Cl胁迫对转基因烟草耐盐性检测、生理生化指标测定和胁迫相关基因的表达分析。结果表明,Na Cl胁迫条件下,转基因烟草叶色失绿程度较野生型的轻,分化情况较野生型的好,生长速度和生根率均较野生型的高;转基因烟草叶绿素和脯氨酸含量及POD和SOD活性均高于野生型烟草;与野生型烟草相比,转基因烟草P5CS、POD、Mn SOD和Gu Zn SOD相对表达水平显著提高。这说明,向日葵HB-12在转基因烟草中的过量表达可抑制叶绿素降解酶的活性,降低叶绿素的分解,还可诱导脯氨酸合成酶基因P5CS及抗氧化相关基因POD、Mn SOD和Gu Zn SOD的上调表达,促进脯氨酸的生物合成,增强POD和SOD的活性,提高烟草抵抗盐害的能力。该研究结果将为进一步探讨植物的耐盐机理及改良作物的耐盐性状奠定基础。Transcription factor HB-12 from sunflower belongs to HD-Zip I transcription factor family and plays an important role in plant stress response. Subcellular localization analysis showed that HB-12 protein was located in the nucleus. At the same time, the plant superexpression vector of HB-12 was constructed and tobacco leaves were transformed by Agrobacterium-mediated transformation and transgenic plants were obtained. The salt tolerance of transgenic tobacco was tested, the physiological and biochemical indexes were determined and the expression of stress related genes were analyzed under Na Cl stress. The results showed that under Na Cl stress, the degree of discoloration in transgenic tobacco leaves was lighter than that of in wild type leaves, and the differentiation of transgenic tobacco was better than that of wild type, the growth rate and rooting rate of transgenic tobacco plants were higher compaired to that of wild type tobacco; The contents of chlorophyll and proline and the activities of POD and SOD were higher than wild type tobacco. The relative gene expression level of transgenic tobacco were significantly higher than those of wild tobacco. These indicated that overexpression of HB-12 in tobacco enhanced its Na Cl tolerance, which would set the foundation for further study on the mechanism of salt tolerance in plants and the improvement of salt-tolerant traits in crops. The overexpression of HB-12 from sunflower in transgenic tobacco could inhibit the activity of chlorophyll degrading enzyme and decrease the decomposition of chlorophyll. The proline synthase gene P5 CS and the antioxidant related gene POD, Mn SOD and Gu Zn SOD were induced to up-regulated expression, promoting the biosynthesis of praline and enhancing the activities of POD and SOD, improving the ability of tobacco to resist salt damage. The results set the foundation for further study in the mechanism of salt tolerance in plants and the improvement of salt-tolerant traits in crops.

关 键 词：向日葵 HB-12 亚细胞定位 过表达 生理生化特性 胁迫相关基因 

分 类 号：Q943.2[生物学—植物学]