低温胁迫下内源性ROS对磷脂酶D活性影响的研究

Effects of Endogenous ROS on Phospholipase D Activity under Low Temperature Stress

低温(4℃,0℃和-4℃)处理高山离子芥(chorispora bungeana),研究低温胁迫引发的内源性ROS对高山离子芥叶中线粒体膜结合态磷脂酶D(Phospholipase D PLD)活性的影响,以期揭示内源性ROS对高山离子芥叶中线粒体膜结合态PLD活性的调控机制。结果表明,在4℃,0℃和-4℃处理下,高山离子芥叶中H2O2的含量和PLD活性均高于空白对照组。在正常条件下,用不同浓度H2O2处理后,PLD活性高于空白对照组。当添加NADPH氧化酶抑制剂(diphenylene iodonium DPI)处理后,在3个温度胁迫下PLD活性低于对照组,在该条件下线粒体膜结合态Ca^(2+)含量高于对照组。当用DPI+CaCl2处理后,PLD活性较添加DPI处理组高,而添加Ca^(2+)的螯合剂EGTA处理后,PLD活性与对照组相比表现出降低,表明低温胁迫下高山离子芥叶中Ca^(2+)参与内源性ROS对PLD活性的调控。

The effects of endogenous ROS on mitochondrial membrane phospholipase D（PLD）activity were studied in chorispora bungeanaleaves at low temperature（4℃,0℃ and-4℃）,which aimed at revealing the mechanism of how endogenous ROS regulating mitochondrial membrane-bound PLD activity.The results indicated that the H_2O_2 content and phospholipase D activity were higher than blank control at 4℃,0℃ and-4℃.The phospholipase D activity were higher than blank control when chorispora bungeana was treated by different concentration of H_2O_2 under normal condition.When treated by DPI（diphenylene iodonium DPI）,an inhibitor of NADPH oxidase,the phospholipase D activity was lower than the control.At the same time,the content of mitochondrial membrane bound Ca^（2＋）content increased than the control at4℃,0℃and-4℃.When treated by DPI＋CaCl_2,the phospholipase D activity was higher than the group with single DPI.And when treated by EGTA as the Chelating agents of Ca^（2＋）,the phospholipase D activity decreased,indicating that Ca^（2＋）took part in regulation of endogenous ROS to mitochondrial membrane bound phospholipase D in chorispora bungeanaleaves.