IWF诱导小麦悬浮细胞产生NO及其与Ca2+的关系

摘要

[目的]以感染叶锈菌的小麦叶片细胞间隙液IWF-260作为激发子,刺激小麦洛夫林10悬浮细胞,探讨悬浮细胞受激发子刺激引发的NO变化情况及其产生的可能分子机制.[方法]采用Greiss试剂法及荧光分子探针DAF-2DA标记法检测胞内NO的动态变化,同时借助药物学试验探讨Ca2+和NO的关系.[结果]IWF-260能诱导小麦悬浮细胞产生NO,NO在IWF-260刺激诱发的细胞死亡过程中发挥重要作用.药物学抑制剂试验证明NOS和NR及胞外钙离子内流均与IWF-260刺激小麦悬浮细胞产生的NO有密切关系,且在NO的产生途径中,NR途径为主要途径.[结论]IWF-260能够诱导小麦悬浮细胞产生NO,NOS、NR及胞外Ca2+内流参与了此过程.%[Objective] Intercellular washing fluid (IWF) obtained from wheat leaves infected by leaf rust contains specific elicitors that was used to stimulate the suspension cell of wheat (cv. Lovrin 10) in order to study the nitric oxide (NO) acting and its possible molecular mechanism. [Method] Greiss reagent and fluorescent NO probe DAF-2DA were used to detecting of dynamic changes of intracellular NO. Pharmacological experiments were carried out to study the signal interactions between calcium (Ca2+) and NO. [Result] IWF-260 could cause the production of NO in wheat suspension cultured cells. It played an important role in the programmed death of wheat suspension cells which was induced by IWF-260. Pharmacological inhibitor experiments proved that NOS and NR might be involved in the production of NO, and the influx of extracellular Ca2+ could regulate the production of NO. NR pathway may be the major route in NO production. [Conclusion] IWF-260 can induce the production of NO in wheat suspension cells, and this process is involved with the NOS, NR and the influx of extracellular Ca2+.