Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, United Kingdom.
Plant Physiol. 2013 Oct;163(2):625-34. doi: 10.1104/pp.113.222901. Epub 2013 Sep 11.
Appropriate stimulus-response coupling requires that each signal induces a characteristic response, distinct from that induced by other signals, and that there is the potential for individual signals to initiate different downstream responses dependent on cell type. How such specificity is encoded in plant signaling is not known. One possibility is that information is encoded in signal transduction pathways to ensure stimulus- and cell type-specific responses. The calcium ion acts as a second messenger in response to mechanical stimulation, hydrogen peroxide, NaCl, and cold in plants and also in circadian timing. We use GAL4 transactivation of aequorin in enhancer trap lines of Arabidopsis (Arabidopsis thaliana) to test the hypothesis that stimulus- and cell-specific information can be encoded in the pattern of dynamic alterations in the concentration of intracellular free Ca(2+) ([Ca(2+)]i). We demonstrate that mechanically induced increases in [Ca(2+)]i are largely restricted to the epidermal pavement cells of leaves, that NaCl induces oscillatory [Ca(2+)]i signals in spongy mesophyll and vascular bundle cells, but not other cell types, and detect circadian rhythms of [Ca(2+)]i only in the spongy mesophyll. We demonstrate stimulus-specific [Ca(2+)]i dynamics in response to touch, cold, and hydrogen peroxide, which in the case of the latter two signals are common to all cell types tested. GAL4 transactivation of aequorin in specific leaf cell types has allowed us to bypass the technical limitations associated with fluorescent Ca(2+) reporter dyes in chlorophyll-containing tissues to identify the cell- and stimulus-specific complexity of [Ca(2+)]i dynamics in leaves of Arabidopsis and to determine from which tissues stress- and circadian-regulated [Ca(2+)]i signals arise.
适当的刺激-反应偶联要求每个信号诱导出特征性的反应,与其他信号诱导的反应不同,并且存在单个信号根据细胞类型启动不同的下游反应的可能性。植物信号中这种特异性是如何编码的尚不清楚。一种可能性是,信息编码在信号转导途径中,以确保刺激和细胞类型特异性反应。钙离子在植物中作为第二信使,对机械刺激、过氧化氢、NaCl 和冷作出反应,也在生物钟计时中起作用。我们使用 GAL4 对拟南芥(Arabidopsis thaliana)增强子陷阱系的水母发光蛋白进行转激活,以检验这样的假设,即刺激和细胞特异性信息可以编码在细胞内游离 Ca(2+)浓度([Ca(2+)]i)动态变化的模式中。我们证明,机械诱导的[Ca(2+)]i增加主要局限于叶片的表皮铺细胞,NaCl 诱导海绵状叶肉和维管束细胞的振荡[Ca(2+)]i信号,但不诱导其他细胞类型,并且仅在海绵状叶肉中检测到[Ca(2+)]i的昼夜节律。我们证明了对触摸、冷和过氧化氢的刺激特异性[Ca(2+)]i动力学,在后两种情况下,这些信号是所有测试细胞类型共有的。在特定的叶片细胞类型中,GAL4 对水母发光蛋白的转激活使我们能够绕过与含有叶绿素的组织中的荧光 Ca(2+)报告染料相关的技术限制,从而确定拟南芥叶片中[Ca(2+)]i动力学的细胞和刺激特异性复杂性,并确定应激和生物钟调节的[Ca(2+)]i信号来自哪些组织。