IWF诱导小麦悬浮细胞H_2O_2迸发及其产生机制初探

【目的】采用具有激发子活性的感染叶锈菌的小麦叶片细胞间隙液IWF-260刺激小麦洛夫林10悬浮细胞,探讨细胞感受激发子刺激引发的H2O2迸发情况及其产生的可能分子机制。【方法】采用化学发光法以及荧光探针DCFHDA、HE来研究H2O2的迸发以及NADPH氧化酶的活化。同时借助药物学试验探讨Ca2+和活性氧的关系。【结果】IWF-260可以引起悬浮细胞H2O2爆发,其在IWF-260刺激诱发的细胞死亡过程中发挥重要作用。而质膜NADPH氧化酶的激活以及胞外钙离子的内流与H2O2爆发有密切关系。【结论】IWF-260能够诱导细胞产生活性氧,质膜NADPH氧化酶和胞外钙离子内流参与了活性氧爆发过程。

Induction of H_2O_2 Burst of Wheat Suspension Cells by IWF and Preliminary Study On Its Productive Mechanism

Objective] Intercelluar washing fluid (IWF-260) of leaf rust - infected wheat leaves ( Lovrin 10) was used to stimulate the suspension cell of Lovrin 10 in order to study the H_2O_2 burst and its productive mechanism. Method] The changes of H2O2 were detected by chemiluminescence method and fluorescent probe DCFHDA staining. The activity of NADPH oxidase was studied by HE staining. Pharmacological experiments were carried out to study the relation between Ca~(2+) and H_2O_2. Result] IWF-260 could cause the outbreak of H_2O_2 in wheat suspension cultured cells. It played an important role in the programmed death of wheat suspension cells induced by IWF-260. NADPH oxidase might be involved in the production of H2O2. The influx of extracellular Ca~(2+) could regulate the production of H_2O_2. Conclusion] IWF can induce H_2O_2 burst of wheat suspension cells and this process is involved with the activation of NADPH oxidase and the influxing of intercelluar Ca~(2+).