过表达小麦锌指型转录因子基因TaZAT8烟草根系差异蛋白的鉴定

Identification of Differential Proteins in Roots of Tobacco with Overexpressing TaZAT8, a Zinc Finger Type Transcription Factor Gene from Wheat

为了在前期研究基础上进一步阐明小麦C2H2型锌指转录因子基因TaZAT8增强植株抵御低磷逆境的分子机制,采用蛋白质组学技术,对正常培养的野生型(WT)与过表达TaZAT8基因烟草株系蛋白表达谱进行了分析。结果表明,与WT相比,过表达株系根系中22个蛋白质的表达发生了显著变化,其中上调表达蛋白为20个,下调表达蛋白2个。然后采用LC-MS/MS质谱技术对差异蛋白进行鉴定,发现上述蛋白分别归属于物质代谢、能量代谢、逆境应答及细胞保护、转录翻译、蛋白质转换和功能未知6个类别。利用生物信息学软件进行亚细胞定位分析,22个差异蛋白主要定位于细胞质、线粒体、过氧化物酶体、细胞核和叶绿体5个不同部位,GO生物过程和分子功能分析表明,上述差异蛋白参与了不同代谢过程或体现不同类别的酶活性。结合上述分析发现,过表达TaZAT8基因通过对物质代谢、能量产生和逆境应答等生物过程的相关蛋白进行调节,为增强植株抵御逆境能力奠定了基础。

In order to further clarify the mechanism of over expression TaZAT8,a wheat C2H2 type zinc finger gene,to enhance the resistance of tobacco plants under P-starvation,the classical proteomics approach was applied to reveal the changes of protein expression between WT and transgenic tobacco plants.The results showed that there were 22 protein spots differentially expressed on 2-DE images,and among them,twenty protein spots were up-regulated and two were down-regulated respectively.Subsequently,these protein spots were identified by LC-MS/MS,which were classified into six functional groups,including primary metabolism,energy generation,stress response and defense,transcription and translation,protein turnover and unclear functions.Subcellular localization analysis indicated that these proteins were mainly located in five organelles,including cytoplasm,mitochondria,peroxisome,nucleus and chloroplast.Go analysis showed that the differentially expressed proteins participated in various metabolism processes or act as enzymes of different categories.Above results showed that over expression TaZAT8 laid the foundation for improving the plant tolerance to stress via regulating proteins involved in biological processes such as substance metabolism,energy production,stress response,etc.