玉米转脂蛋白新成员ZmLTP3的抗盐功能及其上游调控机制研究

项目名称： 玉米转脂蛋白新成员ZmLTP3的抗盐功能及其上游调控机制研究

项目编号： No.31471510

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 食品科学、农学基础与作物学

项目作者： 邹华文

作者单位： 长江大学

项目金额： 90万元

中文摘要： 植物转脂蛋白（LTPs）在非生物胁迫中的作用及其上游调控机制鲜有深入报道。前期研究发现，过量表达ZmPto/ZmPti1的拟南芥抗盐性增强的同时LTP3表达剧增，而拟南芥Pti1突变体中LTP3表达下降，暗示LTP3是Pti1的下游组分且参与抗盐性的调节；转玉米ZmLTP3基因(类LTP3序列)的拟南芥抗盐性提高。本研究拟利用超表达和RNAi技术获得转基因玉米，明确ZmLTP3在玉米中的抗盐功能；克隆ZmLTP3启动子，通过酵母单杂交从玉米cDNA文库中筛选与之作用的转录因子，并经EMSA，ChIP-PCR进行验证，探讨ZmLTP3上游调控组分；分析ZmLTP3在过量和抑制表达ZmPti1的转基因玉米中的表达情况，确定ZmPti1和ZmLTP3的上下游关系；通过ZmPti1对ZmLTP3上游转录因子的酵母双杂交、BiFC及磷酸化检测，初步探讨ZmPti1对ZmLTP3的调控机制。

中文关键词： 玉米；转脂蛋白；抗盐；启动子；转录因子

英文摘要： The functions of plant lipid transfer proteins (LTPs) in abiotic stresses and regulation networks are little reported. In our previous studies, Arabidopsis overexpressing ZmPto/ZmPti1 enhanced resistances to abiotic stresses, meanwhile, the expression level of LTP3 in transgenic lines increased dramatically. The expression of LTP3 in Pti1 mutant lines decreased, indicating LTP3 may be in the downstream of Pti1 signal pathway and participate in the abiotic stresses resistance. Additionaly, Arabidopsis overexpressing maize ZmLTP3, a homologue to LTP3, enhanced the resistance to salt. In this study, overexpression and RNAi methods will be used to transform maize. The functions in salt resistance of ZmLTP3 will be analyzed in transgeic maize. The promoter of ZmLTP3 will be cloned and characterized. Using yeast one-hybrid system, the transcript factors binding to promoter will be isolated from maize cDNA library, and verified by EMSA and ChIP-PCR, clearing the upstream regulation components of ZmLTP3. The expression levels of ZmLTP3 will be analyzed in ZmPti1 overexpression and RNAi maize lines to study the relationship of ZmPti1 and ZmLTP3 in maize. Lastly, yeast one-hybrid system, BiFC and the phosphorylation ability of ZmPti1 to the transcript factors will be analyzed to investigate the regulation mechanisms of ZmPti1 to ZmLTP3.

英文关键词： Maize (Zea mays L.);Lipid transfer protein;Salt resistance;Promoter;Transcription factor