水稻bip130与OSA7蛋白互作的验证及bip130对质膜H+-ATPase活性的影响  

Identification of the interaction between bip130 and OSA7 and the effect of bip130 on plasma membrane H+-ATPase activity in rice

作　　者：张雅芬 刘亚琴 蒋明义[1] ZHANG Yafen;LIU Yaqin;JIANG Mingyi(College of Life Sciences,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学生命科学学院,江苏南京210095

出　　处：《南京农业大学学报》2021年第3期457-467,共11页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31471427)。

摘　　要：[目的]本文旨在验证水稻中bip130(油菜素内酯受体蛋白BR11互作蛋白130)与OSA7(质膜H+-ATP酶7)的相互作用以及ABA处理下bip130对水稻根部质膜H+-ATPase活性的影响。[方法]采用酵母双杂交技术(Y2H)、萤火虫荧光素酶互补成像系统(LCI)、GST pull-down、双分子荧光互补技术(BiFC)验证bip130与OSA7是否相互作用,并用Y2H与LCI验证bip130与OSA7相互作用的区域以及bip130与OSA7同源蛋白的关系。利用RT-qPCR分析ABA处理时bip130与OSA7的上、下游关系。利用bip130突变体通过水稻原生质体瞬时表达体系检测bip130对OSA7活性的影响。[结果]bip130与OSA7的胞质N结构域相互作用,不与5个同源蛋白的胞质N结构域互作。ABA信号途径中bip130位于OSA7的上游;突变体分析结果显示,bip130过表达进一步增强ABA诱导的质膜H+-ATPase活性,而bip130-RNAi阻止ABA诱导的质膜H+-ATPase活性增加。原生质体瞬时表达结果显示在ABA信号途径中bip130可以调控质膜H+-ATPase活性。[结论]bip130与OSA7的胞质N结构域相互作用,且bip130参与调控ABA信号途径中质膜H+-ATPase的活性。[Objectives]This paper was aimed to confirm the interaction between bip130(BR11-interacting protein 130)and OSA7(plasma membrane H+-ATPase 7)and verify that bip130 could regulate the activity of plasma membrane H+-ATPase in rice roots under ABA treatment.[Methods]Yeast two-hybrid technology(Y2H),firefly luciferase complementary imaging system(LCI),GST pull-down and bimolecular fluorescence complementary technology(BiFC)were used to verify whether bip130 interacts with OSA7.Then,Y2H and LCI had been used to identify the interaction region between bip130 and OSA7 and the relationship between bip130 and OSA7 homologous proteins.RT-qPCR had been used to analyze the upstream and downstream relationship between bip130 and OSA7 under ABA treatment.The bip130 mutant materials have been used in the rice protoplast transient expression system to detect whether bip130 could affect the activity of plasma membrane H+-ATPase.[Results]The results indicated that bip130 interacted with the cytoplasmic N-domain of OSA7,but did not interact with the cytoplasmic N-domain of these five homologous proteins.In the ABA signaling pathway,bip130 was located in the upstream of OSA7.Mutant analysis showed that bip130 overexpression could further enhance the activity of ABA-induced plasma membrane H+-ATPase,while bip130-RNAi prevented this process.The analysis of the protoplast transient expression system indicated that bip130 could regulate the activity of plasma membrane H+-ATPase in the ABA signaling pathway.[Conclusions]bip130 interacted with the cytoplasmic N-domain of OSA7 and bip130 participated in the regulation of plasma membrane H+-ATPase activity in ABA signaling pathway.

关 键 词：bip130 OSA7 蛋白互作 ABA 质膜H+-ATPase活性 

分 类 号：Q945[生物学—植物学]