ZmJAZ与ZmMYC2的BiFC互作研究

Interactions Between ZmJAZ and ZmMYC2 Using Bimolecular Fluorescence Complementation Assay

JAZ(jasmonate ZIM-domain)蛋白是茉莉酸(jasmonic acid,JA)信号途径中关键的负调控因子,明确JAZ蛋白和MYC2之间的结合关系对整个JA信号通路至关重要。通过qRT-PCR筛选了在籽粒中较特异表达的ZmJAZ4、生殖器官中高表达的ZmJAZ8以及组成型表达的ZmJAZ12,利用玉米叶肉细胞原生质体瞬时表达体系,通过亚细胞定位和双分子荧光互作(bimolecular fluorescence complementation)研究了这些JAZ蛋白的定位以及是否可以与ZmMYC2相互作用。研究结果发现ZmJAZ4、ZmJAZ8和ZmJAZ12均定位于细胞核,同时BiFC结果显示这些JAZ蛋白都可与ZmMYC2在细胞核中相互作用,证实了ZmJAZ家族蛋白可与JA信号通路关键转录因子ZmMYC2结合的功能,提示它们可通过与ZmMYC2的结合影响其转录调控活性。

JAZ(Jasmonate ZIM-domain)proteins are negative regulators in the JA signaling pathway,clarifying the interaction between JAZ protein and MYC2 is crucial for understanding the JA signaling pathway.qRT-PCR was applied to examine the expressions of ZmJAZ genes,and discovered seed preferentially expressed ZmJAZ4,reproductive organ predominantly expressed ZmJAZ8,and constitutively expressed ZmJAZ12.Then,the subcellular localizations of these AZ proteins and ZmMYC2 were investigated using protoplasts of maize mesophyll cells transiently expressing vector.Besides,the interactions between these JAZ proteins and ZmMYC2 were analyzed by bimolecular fluorescence complementation(BiFC).The results showed that ZmJAZ4,ZmJAZ8,and ZmJAZ12 were located in cell nucleus,and BiFC assay revealed that these JAZ proteins interacted with ZmMYC2 in cell nucleus,confirming that ZmJAZ family proteins had function of binding key transcription factor ZmMYC2 in JA signal pathway,suggesting that ZmMYC2 can regulate its transcriptional activity by binding to ZmMYC2.