《植物生理学报》 2018, 54(7): 1213-1222

NADPH氧化酶在1-戊烯-3-酮诱导的拟南芥早期信号事件中的作用

季亚溪^1,#, 许玲玉^1,#, 焦春阳¹, 高海波², 沈应柏^1,*

¹北京林业大学生物科学与技术学院, 北京100083; ²临沂大学生命科学学院, 山东临沂276005

通信作者：沈应柏；E-mail: ybshen@bjfu.edu.cn

摘　要：

1-戊烯-3-酮(1-penten-3-one)作为重要的植物挥发性信号物质可诱导拟南芥防御反应早期信号的产生。但是拟南芥叶肉细胞响应1-戊烯-3-酮早期信号转导途径尚未见到报道。本研究以拟南芥为材料, 研究了NADPH氧化酶对1-戊烯-3-酮处理后拟南芥早期信号的作用。实验采用非损伤微测技术结合激光共聚焦技术探究了1-戊烯-3-酮诱导的活性氧(ROS)、跨膜离子流信号与NADPH氧化酶之间的关系, 结果表明: 1-戊烯-3-酮处理能够诱导拟南芥叶肉细胞H₂O₂含量增加、Ca²⁺外排和H⁺内流等早期信号事件, NADPH氧化酶抑制剂二联苯碘(DPI)处理后1-戊烯-3-酮诱导的拟南芥叶肉细胞Ca²⁺外排减弱, 胞内钙库抑制剂钌红处理后1-戊烯-3-酮诱导的拟南芥叶肉细胞H⁺内流减弱, 说明1-戊烯-3-酮通过激活NADPH氧化酶诱导了拟南芥叶肉细胞H₂O₂产生, 胞内H₂O₂的积累可能通过激活胞内钙库引起细胞内Ca²⁺浓度增加, 进而引起Ca²⁺的外排和H⁺的内流。

关键词：拟南芥; 1-戊烯-3-酮; NADPH氧化酶; 非损伤微测技术

收稿：2018-04-04   修定：2018-06-12

资助：国家自然科学基金(31270655)、国家基础科学人才培养基金(J1310005)和山东省自然科学基金(31270655)。

Role of NADPH oxidase in early signal events induced by 1-penten-3-one in Arabidopsis thaliana

JI Ya-Xi^1,#, XU Ling-Yu^1,#, JIAO Chun-Yang¹, GAO Hai-Bo², SHEN Ying-Bai^1,*

¹College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China; ²College of Life Science, Linyi University, Linyi, Shandong 276005, China

Corresponding author: SHEN Ying-Bai; E-mail: ybshen@bjfu.edu.cn

Abstract:

1-penten-3-one, an important plant volatile, can induce early signal events in Arabidopsis thaliana defense against abiotic stress. However, the research on how early signal occurs in plant response to 1-penten-3-one has not been reported. In this study, A. thaliana was used as the material to study the effect of NADPH oxidase on early signal patterns in A. thaliana under 1-penten-3-one treatment. The non-invasive microassay technique combined with laser confocal technology was used to explore the internal relationship between reactive oxygen species, transmembrane ions fluxes, and NADPH oxidase induced by 1-penten-3-one. The results show that 1-penten-3-one could induce early signal events such as H₂O₂ production and accumulation, Ca²⁺ efflux, and H⁺ influx in A. thaliana mesophyll cells. Moreover, Ca²⁺ efflux induced by 1-pentene-3-one was significantly suppressed in the mesophyll cells of A. thaliana pretreated with diphenyleneiodonium, an inhibitor of NADPH oxidase; after an intracellular calcium inhibitor ruthenium red treatment, H⁺ influx elicited by 1-penten-3-one in A. thaliana mesophyll cells was suppressed as well. These data suggest that 1-penten-3-one could induce H₂O₂ accumulation in A. thaliana mesophyll cells through activating NADPH oxidase. The accumulation of intracellular H₂O₂ was involved in elevating intracellular Ca²⁺ concentration through the activation of intracellular calcium stores, which in turn led to Ca²⁺ efflux and H+ influx.

Key words: Arabidopsis thaliana; 1-penten-3-one; NADPH oxidase; non-invasive microassay