González-García Mary Paz, Rodríguez Dolores, Nicolás Carlos, Rodríguez Pedro Luis, Nicolás Gregorio, Lorenzo Oscar
Departamento de Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología, Universidad de Salamanca, 37007 Salamanca, Spain.
Plant Physiol. 2003 Sep;133(1):135-44. doi: 10.1104/pp.103.025569.
FsPP2C1 was previously isolated from beech (Fagus sylvatica) seeds as a functional protein phosphatase type-2C (PP2C) with all the conserved features of these enzymes and high homology to ABI1, ABI2, and PP2CA, PP2Cs identified as negative regulators of ABA signaling. The expression of FsPP2C1 was induced upon abscisic acid (ABA) treatment and was also up-regulated during early weeks of stratification. Furthermore, this gene was specifically expressed in ABA-treated seeds and was hardly detectable in vegetative tissues. In this report, to provide genetic evidence on FsPP2C1 function in seed dormancy and germination, we used an overexpression approach in Arabidopsis because transgenic work is not feasible in beech. Constitutive expression of FsPP2C1 under the cauliflower mosaic virus 35S promoter confers ABA insensitivity in Arabidopsis seeds and, consequently, a reduced degree of seed dormancy. Additionally, transgenic 35S:FsPP2C1 plants are able to germinate under unfavorable conditions, as inhibitory concentrations of mannitol, NaCl, or paclobutrazol. In vegetative tissues, Arabidopsis FsPP2C1 transgenic plants show ABA-resistant early root growth and diminished induction of the ABA-response genes RAB18 and KIN2, but no effect on stomatal closure regulation. Seed and vegetative phenotypes of Arabidopsis 35S:FsPP2C1 plants suggest that FsPP2C1 negatively regulates ABA signaling. The ABA inducibility of FsPP2C1 expression, together with the transcript accumulation mainly in seeds, suggest that it could play an important role modulating ABA signaling in beechnuts through a negative feedback loop. Finally, we suggest that negative regulation of ABA signaling by FsPP2C1 is a factor contributing to promote the transition from seed dormancy to germination during early weeks of stratification.
FsPP2C1 先前是从山毛榉(欧洲山毛榉)种子中分离出来的,它是一种功能性 2C 型蛋白磷酸酶(PP2C),具有这些酶的所有保守特征,并且与 ABI1、ABI2 和 PP2CA 高度同源,这些 PP2C 被确定为脱落酸(ABA)信号传导的负调节因子。FsPP2C1 的表达在脱落酸(ABA)处理后被诱导,并且在层积处理的最初几周也上调。此外,该基因在 ABA 处理的种子中特异性表达,在营养组织中几乎检测不到。在本报告中,为了提供关于 FsPP2C1 在种子休眠和萌发中功能的遗传证据,我们在拟南芥中采用了过表达方法,因为在山毛榉中进行转基因工作不可行。在花椰菜花叶病毒 35S 启动子控制下组成型表达 FsPP2C1 会使拟南芥种子对 ABA 不敏感,从而导致种子休眠程度降低。此外,转基因 35S:FsPP2C1 植物能够在不利条件下萌发,如甘露醇、NaCl 或多效唑的抑制浓度下。在营养组织中,拟南芥 FsPP2C1 转基因植物表现出对 ABA 抗性的早期根系生长,并且 ABA 响应基因 RAB18 和 KIN2 的诱导减弱,但对气孔关闭调节没有影响。拟南芥 35S:FsPP2C1 植物的种子和营养表型表明 FsPP2C1 负调节 ABA 信号传导。FsPP2C1 表达的 ABA 诱导性,以及转录本主要在种子中的积累,表明它可能通过负反馈环在山毛榉坚果中调节 ABA 信号传导中发挥重要作用。最后,我们认为 FsPP2C1 对 ABA 信号传导的负调节是在层积处理的最初几周促进种子从休眠向萌发转变的一个因素。