拟南芥CARK3与RCAR12相互作用的研究

植物激素ABA是植物应对非生物胁迫的响应因子,广泛参与了植物生长发育的调控.ABA结合其受体后抑制PP2Cs的磷酸酶活性,释放SnRK2s,从而启动ABA信号转导途径.本文采用酵母双杂交系统,GST-pull down技术研究蛋白质激酶CARK3与ABA受体RCAR12在体外的相互作用,结果显示CARK3和RCAR12共转入酵母后,在三缺平板上能够正常生长;经His-Tag抗体检测,CARK3与RCAR12出现明显且单一的条带.同时,采用双分子荧光互补实验研究CARK3与RCAR12在体内的相互作用,结果显示CARK3与RCAR12共转化烟草后能观察到较强的荧光.体内外实验表明,CARK3与RCAR12存在明显的相互作用,CARK3可能通过直接磷酸化ABA受体RCAR12来调控ABA信号途径的生理应答.

An Interaction of ABA Receptor RCAR12 with CARK3 in Arabidopsis thaliana

The phytohormone abscisic acid (ABA) is crucial for plant development and response to abiotic stress, including drought. ABA perceived by the ABA receptors, PYR/PYL/RCAR, which bound to ABA, and then recruit Protein phosphatase 2C (PP2C) to inhibit the PP2C acticity, thereby activating SNF1-RELATED PROTEIN KINASE2 (SnRK2s). Here, CARK3 and RCAR12 interact in yeast two-hybrid system and GST-pull down in vitro. Bimolecular fluorescence complementation (BiFC) was used to illuminate the interaction between CARK3 and RCAR12 in Nicotiana benthamiana in vivo. We could confer that CARK3 indeed interacts with RCAR12 in present study and the phosphorylation modulates the physiological response of the core ABA signaling pathway. Our results provide the foundation for further understanding of the regulatory network of phosphorylation of ABA signaling pathways core components.