核黄素对烟草悬浮细胞活性氧和防卫反应的激活作用  

Activation of Reactive Oxygen Species and Defense Responses in Tobacco Cells Treated with Riboflavin

作　　者：刘菲[1] 魏芳芳[1] 王蕾[1] 刘辉[1] 梁元存[1] 刘爱新[1] 

机构地区：[1]山东农业大学植物保护学院,山东泰安271018

出　　处：《中国农业科学》2009年第12期4175-4181,共7页Scientia Agricultura Sinica

基　　金：山东省教育厅项目(J08LF06)

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】1mmol·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】1mmol·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。【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 transcriptase-polymerase chain reaction） and HPLC（high performance liquid chromatography） techniques.【Result】 It was identified that a series of defense-related responses were induced in tobacco suspension cells after treated with riboflavin,such as an oxidative burst,resulting in alkalization of the extracellular medium,activated the expression of four defense-related genes,and the scopoletin accumulation.And all the tested three inhibitors were shown in suppressing the riboflavin triggered defense responses at difference levels.【Conclusion】 These results suggest that riboflavin triggers plant basal defense signal pathway which are also concerned with calcium signal pathway and protein phosphorylation in tobacco cell suspensions as an elicitor.

关 键 词：核黄素 烟草 活性氧 防卫反应 

分 类 号：S572[农业科学—烟草工业]