《植物生理学报》 2011, 47(1): 85-90

机械刺激诱导的烟草悬浮细胞一氧化氮产生途径及其与Ca²⁺和钙调素的关系

柯学^1,2, 李忠光^2,3,4, 刘娴^2,3,4, 龚明^2,3,4,*

1 中国农业大学水利与土木工程学院, 北京 10 008 3; 云南师范大学2 生命科学学院, 3 生物能源持续开发利用教育部工程研 究中心, 4 云南省生物质能与环境生物技术重点实验室, 昆明 650092

通信作者：龚明；E-mail: gongming63@163.com；Tel: 0871-5516516

摘　要：

通过提高摇床转速对烟草细胞施加机械刺激(MS)可诱导其胞内一氧化氮(NO)的快速产生和一氧化氮合酶(NOS)活性 的提高, 这种 MS 诱导的 NO 产生可被 NO 清除剂 cPTIO 和 NOS 抑制剂 L-NMMA 显著抑制。此外, Ca2+ 螯合剂 EGTA、质 膜 Ca2+ 通道阻断剂 La3+、胞内 Ca2+ 通道拮抗剂钌红, 以及钙调素抑制剂 CPZ 和 TFP 预处理均不同程度地抑制了机械刺激 诱导的烟草细胞NO 的产生, 而机械刺激过程中钙调素活性显著上升并与NOS活性和NO 含量的变化相一致。这些结果暗 示着(类)NOS酶催化的精氨酸依赖途径可能是机械刺激诱发烟草细胞NO产生的主要途径, Ca2+/CaM可能通过调节(类)NOS 活性来调控 NO 的产生。

关键词：机械刺激; 一氧化氮; 一氧化氮合酶; 钙; 钙调素; 烟草细胞

收稿：2010-09-15   修定：2010-09-27

资助：国家自然科学基金(30460016) 和云南省教育厅基金(07Z11860, 09Y0145)。

Pathways of Mechanical Stimulation-Induced Nitric Oxide Production and Their Relationship with Ca²⁺ and Calmodulin in Tobacco (Nicotiana tobacum L.) Suspension Culture Cells

KE Xue^1,2, LI Zhong-Guang^2,3,4, LIU Xian^2,3,4, GONG Ming^2,3,4,*

¹College of Water Conservancy & Civil Engineering, China Agricultural University, Beijing 100083, China; ²School of Life Sciences,³Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education,⁴Key Laboratory of Biomass Energy and Environmental Biotechnology, Yunnan Province, Yunnan Normal University, Kunming 650092,China

Corresponding author: GONG Ming; E-mail: gongming63@163.com; Tel: 0871-5516516

Abstract:

Application of mechanical stimulation (MS) on tobacco (Nicotiana tobacum) suspension culture cells led to rapid production of intracellular nitric oxide (NO) and increased activity of nitric oxide synthase (NOS), but this MS-induced NO production could be significantly inhibited by the NO scavenger cPTIO and NOS inhibitor L-NMMA. In addition, the Ca²⁺ specific chelator EGTA, plasma membrane Ca²⁺ channel blocker La³⁺,intracellular Ca²⁺ channel antagonist ruthenium red, and the calmodulin inhibitors chlorpromazine and trifluoperazine pretreatments all led to the inhibition of the MS-induced NO production to some extent; on the other hand, the MS treatment raised calmodulin activity in the tobacco cells, being consistent with the change of NO content and NOS activity. All this results suggested that the arginine-dependent pathway catalyzed by NOS-like enzyme could be main source for this MS-induced NO production and Ca²⁺/calmodulin was involved in the regulation of the NO production by control of NOS activity.

Key words: mechanical stimulation; nitric oxide; nitric oxide synthase; calcium; calmodulin; tobacco suspension cells