State Key Laboratory of Plant Physiology and Biochemistry, Beijing, P. R. China.
National Plant Gene Research Center, College of Biological Sciences, China Agricultural University, Beijing, P. R. China.
J Exp Bot. 2018 Mar 24;69(7):1569-1582. doi: 10.1093/jxb/erx488.
Strawberry (Fragaria×ananassa) is a model plant for studying non-climacteric fruit ripening regulated by abscisic acid (ABA); however, its exact molecular mechanisms are yet not fully understood. In this study, a predicted leu-rich repeat (LRR) receptor-like kinase in strawberry, red-initial protein kinase 1 (FaRIPK1), was screened and, using a yeast two-hybrid assay, was shown to interact with a putative ABA receptor, FaABAR. This association was confirmed by bimolecular fluorescence complementation and co-immunoprecipitation assays, and shown to occur in the nucleus. Expression analysis by real-time PCR showed that FaRIPK1 is expressed in roots, stems, leaves, flowers, and fruit, with a particularly high expression in white fruit at the onset of coloration. Down-regulation of FaRIPK1 expression in strawberry fruit, using Tobacco rattle virus-induced gene silencing, inhibited ripening, as evidenced by suppression of ripening-related physiological changes and reduced expression of several genes involved in softening, sugar content, pigmentation, and ABA biosynthesis and signaling. The yeast-expressed LRR and STK (serine/threonine protein kinase) domains of FaRIPK1 bound ABA and showed kinase activity, respectively. A fruit disc-incubation test revealed that FaRIPK1 expression was induced by ABA and ethylene. The synergistic action of FaRIPK1 with FaABAR in regulation of strawberry fruit ripening is discussed.
草莓( Fragaria ×ananassa )是研究脱落酸(ABA)调控非跃变型果实成熟的模式植物;然而,其确切的分子机制尚不完全清楚。本研究在草莓中筛选到一个预测的富含亮氨酸重复(LRR)受体样激酶,即红色初始蛋白激酶 1(FaRIPK1),并通过酵母双杂交实验证实其与一个假定的 ABA 受体 FaABAR 相互作用。该关联通过双分子荧光互补和共免疫沉淀实验得到证实,并发生在细胞核中。实时 PCR 表达分析表明 FaRIPK1 在根、茎、叶、花和果实中表达,在果实开始着色时的白果中表达水平特别高。利用烟草脆裂病毒诱导的基因沉默下调草莓果实中 FaRIPK1 的表达,可抑制果实成熟,表现为成熟相关生理变化受到抑制,参与软化、糖含量、色素形成和 ABA 生物合成及信号转导的几个基因表达降低。酵母表达的 FaRIPK1 的 LRR 和 STK(丝氨酸/苏氨酸蛋白激酶)结构域分别结合 ABA 和显示激酶活性。果实圆盘孵育试验表明 FaRIPK1 的表达受 ABA 和乙烯的诱导。讨论了 FaRIPK1 与 FaABAR 协同调控草莓果实成熟的作用。