核黄素对烟草悬浮细胞活性氧和防卫反应的激活作用.pdfVIP

中国农业科学 2009,42(12):4175-4181 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2009.12.007 核黄素对烟草悬浮细胞活性氧和防卫反应的激活作用 刘 菲，魏芳芳，王 蕾，刘 辉，梁元存，刘爱新 （山东农业大学植物保护学院，山东泰安 271018 ） 摘要：【目的】研究核黄素处理烟草悬浮细胞后，激活的氧迸发和防卫反应，以及钙信号和蛋白激酶对这些 事件的影响。【方法】1 mmol·L-1 核黄素处理烟草悬浮细胞，用半定量RT-PCR（reverse transcriptase-polymerase chain reaction）和高效液相色谱（high performance liquid chromatography，HPLC）等方法测定一系列防卫 反应，并用钙离子通道抑制剂LaCl3、钙离子螯合剂EGTA和一种丝氨酸/苏氨酸蛋白激酶抑制剂staurosporine 处 理细胞，测定防卫反应的变化。【结果】1 mmol·L-1 核黄素处理悬浮细胞后，产生了氧迸发、导致培养基质碱性化、 增强4种防卫基因 PR （pathogenesis-related）-1a、PR-1b、PAL （phenylalanine ammonia lyase）和LPO （lignin peroxidase）表达以及 scopoletin 的积累；LaCl3、EGTA 和 staurosporine 可以不同程度地抑制这些防卫反应。 【结论】核黄素能够激活烟草悬浮细胞的氧迸发和防卫反应，在信号转导中钙信号和蛋白质磷酸化参与了这些过 程。 关键词：核黄素；烟草；活性氧；防卫反应 Activation of Reactive Oxygen Species and Defense Responses in Tobacco Cells Treated with Riboflavin LIU Fei, WEI Fang-fang, WANG Lei, LIU Hui, LIANG Yuan-cun, LIU Ai-xin (College of Plant Protection, Shandong Agricultural University, Tai’an 271018, Shandong) Abstract: 【Objective 】 To study the triggering defense responses in tobacco suspension cells after treated with riboflavin, reactive oxygen species (ROS) and defense-related responses were investigated together with their related effects involving in calcium signals and protein kinases. 【Method 】 Tobacco suspension cells were treated with 1 mmol·L-1 riboflavin or pretreated with inhibitors including calcium surrogate La3+, calcium chelator EGTA and staurosporine which is an inhibitor of serine/threonine protein kinase. A series of dynamic defense responses were then studied with several methods, including semiquantitative RT-PCR (reverse