Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Science, Hebei Normal University, Shijiazhuang 050024, China.
Plant J. 2012 Jun;70(6):1056-69. doi: 10.1111/j.1365-313X.2012.04969.x. Epub 2012 Apr 13.
An increased concentration of cytosolic calcium ions (Ca²⁺) is an early response by plant cells to heat shock. However, the molecular mechanism underlying the heat-induced initial Ca²⁺ response in plants is unclear. In this study, we identified and characterized a heat-activated Ca²⁺-permeable channel in the plasma membrane of Arabidopsis thaliana root protoplasts using reverse genetic analysis and the whole-cell patch-clamp technique. The results indicated that A. thaliana cyclic nucleotide-gated ion channel 6 (CNGC6) mediates heat-induced Ca²⁺ influx and facilitates expression of heat shock protein (HSP) genes and the acquisition of thermotolerance. GUS and GFP reporter assays showed that CNGC6 expression is ubiquitous in A. thaliana, and the protein is localized to the plasma membrane of cells. Furthermore, it was found that the level of cytosolic cAMP was increased by a mild heat shock, that CNGC6 was activated by cytosolic cAMP, and that exogenous cAMP promoted the expression of HSP genes. The results reveal the role of cAMP in transduction of heat shock signals in plants. The correlation of an increased level of cytosolic cAMP in a heat-shocked plant with activation of the Ca²⁺ channels and downstream expression of HSP genes sheds some light on how plants transduce a heat stimulus into a signal cascade that leads to a heat shock response.
细胞溶质钙离子(Ca²⁺)浓度的增加是植物细胞对热激的早期响应。然而,植物中热诱导初始 Ca²⁺响应的分子机制尚不清楚。在这项研究中,我们使用反向遗传学分析和全细胞膜片钳技术,鉴定并表征了拟南芥根原生质体质膜中热激活的 Ca²⁺渗透性通道。结果表明,拟南芥环核苷酸门控离子通道 6(CNGC6)介导热诱导的 Ca²⁺内流,并促进热休克蛋白(HSP)基因的表达和耐热性的获得。GUS 和 GFP 报告基因分析表明,CNGC6 在拟南芥中广泛表达,并且该蛋白定位于细胞的质膜上。此外,研究发现轻度热激会增加细胞溶质 cAMP 的水平,cAMP 激活 CNGC6,外源性 cAMP 促进 HSP 基因的表达。这些结果揭示了 cAMP 在植物中热激信号转导中的作用。热激植物中细胞溶质 cAMP 水平的增加与 Ca²⁺通道的激活以及 HSP 基因的下游表达相关,这为植物如何将热刺激转导成导致热激反应的信号级联提供了一些线索。