植物细胞中蓝光诱导ROS生成的识别和亚细胞定位检测

以2’,7’-二氯二氢荧光素二乙酯(dichlorofluorescein diacetate,H2DCF-DA)为荧光探针孵育拟南芥叶表皮条,利用荧光光谱和激光共聚焦扫描显微技术,对高辐照蓝光诱导下叶肉细胞活性氧(reactive oxygen spe-cies,ROS)的生成,进行了分子识别和亚细胞定位检测。结果表明:植物细胞在蓝光诱导下,可以产生大量的ROS。过氧化氢酶清除实验表明:高辐照蓝光诱导产生的ROS,主要成分是H2O2,并且主要定位在叶绿体和细胞膜上。

Identification and Detection of Subcellular Localization of High-fiuence Blue Light-induced ROS Generation in Plant Cell

In this study,using dichlorofluorescein diacetate(H2DCF-DA) as a probe to incubate the leaf strips of Arabidopsis,we investigated the molecular identification and subcellular localization of reactive oxygen species(ROS) generation in mesophyllic cells of leaf strips induced by high-fluence blue light under a laser scanning confocal microscope and luminescence spectrometer.Results demonstrated that a large amount of ROS production labeled by the fluorescence intensity of dichlorofluorescein(DCF) was observed in mesophyllic cells of leaf strips after treatment with high-fluence blue light.Moreover,the scavenger experiments of catalase indicated that H2O2 is the major component of ROS generation in mesophyllic cells.H2O2 generation induced by 520 Lx high-fluence blue light was mainly localized at chloroplasts and cell membrane.