番茄SIZ1-like1基因的克隆与功能  

Cloning and function study of tomato SUMO E3 ligase SIZ1-like1 gene

作　　者：张俊芳[1] 唐晓凤[1] 李欲翔[1] 邓恒[1] 高兰阳[1] 刘永胜[1] 

机构地区：[1]四川大学生命科学学院,水力学与山区河流开发保护国家重点实验室,成都610064

出　　处：《应用与环境生物学报》2015年第3期406-412,共7页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(90717110);国家高技术研究发展计划(863计划;2007A A10Z122);国家杰出青年科学基金项目(30825030)资助~~

摘　　要：SUMO(Small ubiquitin-related modifi er)化修饰是植物体内一种重要的蛋白质功能调节方式.它调控植物细胞中蛋白的降解与定位,植物抗性及激素调节等.SIZ1是SUMO的E3连接酶并在SUMO化中起重要作用.为了解SIZ1基因在番茄中的功能,成功克隆番茄(Solanum lycopersicum)SIZ1基因(SIZ1-like1)并构建番茄SIZ1-like1RNA干涉和过表达载体后,通过农杆菌介导转入野生型番茄,成功获得6个独立的转基因阳性植株.荧光定量PCR(Real-time PCR)分析野生型番茄中SIZ1-like1基因的组织表达特异性,发现SIZ1-like1在植物的各个组织中都有表达.构建SIZ1-like1黄色荧光蛋白融合表达载体,通过对SIZ1-like1融合黄色荧光蛋白转基因番茄的根尖进行荧光显微观察,证实番茄SIZ1蛋白定位于细胞核.干旱胁迫实验分析显示,过表达转基因植株抗旱性强于野生型,且脯氨酸含量是野生型的3倍左右,而RNAi植株抗旱能力则较弱.因此SIZ1基因对番茄抗旱起到了正调控作用.Sumoylation is an important way to regulate post-translation of proteins in plant cells, including protein degradation and location, responses to environmental stresses and hormones. SIZ1 is a SUMO （small ubiquitin-related modifier） E3 ligase that plays a vital role in sumoylation. In order to illustrate the function of SIZ1 gene in tomato, this research cloned SIZ1 gene from tomato, constructed SIZI-likel-PBI121 over-expression and SIZl-likel-pSKint RNA interference fusion expression vectors, and transformed them into wild type tomato using ,4grobacterium-mediated transformation to successfully obtain six positive transgenic plants. The Real-Time PCR experiment indicated that SIZl-likel expressed in all growth stages of tomato. SIZI-likel-YFP fusion expression vector was constructed and transformed into tomato. The results of microscopic observation revealed that SIZl-likel was located in the nuclei. Drought stress indicated that transgenic plants with overexpressed SIZ1- likel had stronger tolerance compared to the wild type, with proline content about threefold of the latter; however, the SIZ1- likelRNA interference plants had less tolerance. These results demonstrated the positive regulation ofSIZ1 gene in tomato.

关 键 词：番茄 SIZ1-like1 基因克隆 抗旱 亚细胞定位 正调控 

分 类 号：S641.2[农业科学—蔬菜学]