Reactive Oxygen Species Production and Stimulated Endocytosis in Tobacco BY-2 Cells Treated with Erwinia carotovora Culture Filtrate
Reactive Oxygen Species Production and Stimulated Endocytosis in Tobacco BY-2 Cells Treated with Erwinia carotovora Culture Filtrate
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DOI：
10.1508/cytologia.83.289
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发表时间：
2018-09
期刊：
影响因子：
1
通讯作者：
Y. Hirakawa;S. Hasezawa;T. Higaki
中科院分区：
文献类型：
--
作者：
Y. Hirakawa;S. Hasezawa;T. Higaki
我们利用烟草BY-2细胞建立了有效的致病信号诱导细胞死亡的实验体系，并培养了一种植物病原菌--胡萝卜欧文氏菌(fifi)。利用该实验系统，对fi诱导的细胞死亡过程中细胞骨架和空泡的变化进行了详细的表征。然而，fiLtrate引发的初始事件在很大程度上是未知的。在这项研究中，我们检测了胡萝卜型埃希氏菌fi液对BY-2细胞产生活性氧(ROS)和内吞刺激的影响。fi处理后2分钟内可观察到一过性ROS的产生。用钙离子螯合剂BAPTA或蛋白激酶抑制剂K252a处理后，fi可剂量依赖性地抑制ROS的产生，提示ROS的产生依赖于flUX中的钙离子或蛋白磷酸化。此外，fi还可在30min内促进内吞标志物FM4-的内化，提示fi可刺激细胞内吞作用。这些结果表明，fi诱导BY-2细胞产生ROS和随后的内吞刺激是诱导其产生ROS的初始事件。
Summary We previously established an experimental system for efficient pathogenic signal induced-cell death using tobacco BY-2 cells and culture filtrate of a plant pathogenic bacterium, Erwinia carotovora . Using this experimental system, cytoskeletal and vacuolar changes during the process of filtrate-induced cell death were char-acterized in detail. However, the initial events induced by the filtrate were largely unknown. In this study, reactive oxygen species (ROS) production and endocytosis stimulation were examined in BY-2 cells treated with the filtrate of E. carotovora . Transient ROS production within 2 min was observed after the filtrate treatment. Treatment with the Ca 2+ chelator BAPTA or the protein kinase inhibitor K252a significantly inhibited ROS production in a dose-dependent manner, suggesting that ROS production depends on Ca 2+ influx or protein phosphorylation. In addition, internalization of the endocytic marker FM4-64 was promoted by filtrate treatment within 30 min, suggesting that the filtrate treatment stimulated endocytosis. These results showed that ROS production and subsequent endocytosis stimulation are the initial events induced by culture filtrate of E. carotovora in BY-2 cells.