甘蓝型油菜钙依赖蛋白激酶6(BnaCPK6)的定位与互作蛋白分析  

Analysis of Localization and Interacting Proteins of Calcium-Dependent Protein Kinase 6(BnaCPK6) in Brassica napus

作　　者：王伟欢 刘舞贞 杨博 张翰风 邓民 李竞[1] 江元清[1] Wang Weihuan;Liu Wuzhen;Yang Bo;Zhang Hanfeng;Deng Min;Li Jing;Jiang Yuanqing(State Key Laboratory of Crop Stress Biology for Arid Areas, College of Life Sciences, Northwest Agriculture & Forestry University, Yangling, 712100)

机构地区：[1]西北农林科技大学生命科学学院旱区作物逆境生物学国家重点实验室

出　　处：《基因组学与应用生物学》2019年第5期2104-2109,共6页Genomics and Applied Biology

基　　金：国家自然科学基金(31471153)资助

摘　　要：为了探究甘蓝型油菜中钙依赖蛋白激酶(calcium dependent protein kinase, CPK)在植物对逆境响应中的作用和机制,同时为油菜品质的升级改良发掘新的基因资源,开展了对于BnaCPK6研究的分子生物学试验。首先,通过在本氏烟草中瞬时表达BnaCPK6与GFP融合蛋白来检测它的亚细胞定位情况;其次,利用双分子荧光互补(Bimolecular fluorescence complementation)试验来检测BnaCPK6与ABA信号通路中转录因子BanABF1/3/4、BnaABI5、BnaAREB3的相互作用情况。结果显示BnaCPK6具有典型的钙依赖蛋白激酶特征,N端具有潜在的棕榈酰化和豆蔻酰化位点,并且与AtCPK6在进化上有着很高的同源性。亚细胞定位的结果发现BnaCPK6主要分布于细胞膜和细胞核中。同时,双分子荧光互补试验还发现BnaCPK6与调控ABA信号转导的关键转录因子BnaABF3/4、BnaABI5以及BnaAREB3之间存在相互作用。本研究为进一步研究BnaCPK6在ABA信号通路中的作用提供了依据。In order to investigate the role and mechanism of calcium-dependent protein kinase(CPK) of Brassica napus in the plant response to adversity, and also discover the new genetic resource for the upgrade and improvement of rapeseed quality, we carried out molecular biological experiments on BnaCPK6. Firstly, subcellular localization was examined by transient expression Bna CPK6-GFP fusion proteins in Nicotiana benthamiana. Then,the interaction between BnaCPK6 and transcription factors BanABF1/3/4, BnaABI5 and BnaAREB3 in ABA signaling pathway was detected by bimolecular fluorescence complementation assay. The results showed that BnaCPK6 had typical characteristics of calcium-dependent protein kinases, N-terminal had potential palmitoylation and myristoylation sites, and had high evolutionary homology with AtCPK6. Subcellular localization revealed that BnaCPK6 was distributed mainly in cell membrane and nuclei. At the same time, the interaction of the key transcription factors BnaABF3/4, BnaABI5 and BnaAREB3 that regulated ABA signal transduction with BnaCPK6 were confirmed in bimolecular fluorescence complementation assay. In conclusion,our research would provide the basis for further study on the role of BnaCPK6 in the ABA signaling pathway.

关 键 词：钙依赖蛋白激酶 油菜 脱落酸 转录因子 

分 类 号：S565.4[农业科学—作物学]