论文部分内容阅读
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.
SUMO (Small ubiquitin-related modifier) is an important protein function regulation in plants, which regulates the degradation and localization of proteins in plant cells, plant resistance and hormone regulation, etc. SUM1 is SUMO’s E3 ligase Plays an important role in SUMO.In order to understand the function of SIZ1 gene in tomato, we successfully cloned the SIZ1-like1 gene of tomato (Solanum lycopersicum) and constructed the tomato SIZ1-like1 RNA interference and overexpression vector, Six wild-type transgenic plants were successfully transformed into wild-type tomato.The expression specificity of SIZ1-like1 gene in wild-type tomato was analyzed by Real-time PCR and it was found that SIZ1-like1 was expressed in various tissues of plants SIZ1-like1 yellow fluorescent protein fusion expression vector was constructed and SIZ1-like1 fusion yellow fluorescent protein transgenic root tips were observed by fluorescence microscopy to confirm the tomato SIZ1 protein localized in the nucleus.Experimental analysis of drought stress The expression of transgenic plants was stronger than that of the wild type, and proline content was about 3 times that of the wild type, while the RNAi plants had weaker drought tolerance ability. Therefore, SIZ1 Because of the drought tomato played a positive regulatory role.