Sun Qing-Peng, Guo Yi, Sun Ying, Sun Da-Ye, Wang Xiao-Jing
Guangdong Key Laboratory of Biotechnology for Plant Development, South China Normal University, Guangzhou, 510631, China.
J Plant Res. 2006 Jul;119(4):343-50. doi: 10.1007/s10265-006-0279-x. Epub 2006 May 18.
The changes in cytosolic Ca2+ levels play important roles in the signal transduction pathways of many environmental and developmental stimuli in plants and animals. We demonstrated that the increase in cytosolic free Ca2+ concentration ([Ca2+]cyt) of Arabidopsis thaliana leaf cells was induced by exogenous application of jasmonic acid (JA). The elevation of [Ca2+]cyt was detected within 1 min after JA treatment by the fluorescence intensity using laser scanning confocal microscopy, and the elevated level of fluorescence was maintained during measuring time. With pretreatment of nifedipine (Nif), a nonpermeable L-type channel blocker, the fluorescence of [Ca2+]cyt induced by JA was inhibited in a dose-dependent manner. In contrast, verapamil, another L-type channel blocker, had no significant effect. Furthermore, Nif repressed JA-induced gene expression of JR1 but verapamil did not. JA-induced gene expression could be mimicked by higher concentration of extracellular Ca2+. W-7 [N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide], an antagonist of calmodulin (CaM), blocked the JA induction of JR1 expression while W-5 [N-(6-aminohexyl)-1-naphthalenesulfonamide], its inactive antagonist, had no apparent effect. These data provide the evidence that the influx of extracellular Ca2+ through Nif sensitive plasma membrane Ca2+ channel may be responsible for JA-induced elevation of [Ca2+]cyt and downstream gene expression, CaM may be also involved in JA signaling pathway.
胞质Ca2+水平的变化在植物和动物许多环境及发育刺激的信号转导途径中发挥着重要作用。我们证明,外源施加茉莉酸(JA)可诱导拟南芥叶细胞胞质游离Ca2+浓度([Ca2+]cyt)升高。利用激光扫描共聚焦显微镜通过荧光强度检测到,JA处理后1分钟内[Ca2+]cyt升高,且在测量期间荧光升高水平得以维持。用非通透性L型通道阻滞剂硝苯地平(Nif)预处理后,JA诱导的[Ca2+]cyt荧光呈剂量依赖性受到抑制。相比之下,另一种L型通道阻滞剂维拉帕米则无显著影响。此外,Nif抑制JA诱导的JR1基因表达,而维拉帕米则无此作用。较高浓度的细胞外Ca2+可模拟JA诱导的基因表达。钙调蛋白(CaM)拮抗剂W - 7 [N -(6 - 氨基己基)- 5 - 氯 - 1 - 萘磺酰胺]可阻断JA对JR1表达的诱导作用,而其无活性拮抗剂W - 5 [N -(6 - 氨基己基)- 1 - 萘磺酰胺]则无明显作用。这些数据证明,细胞外Ca2+通过Nif敏感的质膜Ca2+通道内流可能是JA诱导的[Ca2+]cyt升高及下游基因表达的原因,CaM可能也参与JA信号通路。