FNOCT as a fluorescent probe for in vivo localization of nitric oxide distribution in tobacco roots

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	FNOCT as a fluorescent probe for in vivo localization of nitric oxide distribution in tobacco roots
	Vandana, S.
刊名	Plant Physiology and Biochemistry
	2012
卷号	59 页码:80-89
ISSN号	0981-9428
中文摘要	The nitric oxide-specific fluorescent probe Fluorescent Nitric Oxide Cheletropic Trap (FNOCT) 8a was applied to intact tobacco (Nicotiana tabacum cv. Samsun) roots to detect sites of nitric oxide formation and NO distribution. Three week old tobacco seedlings were gently removed from the sand culture pots with intact roots and transferred to small Petri dishes, whose base was replaced by a thin coverslip. Intact roots were subjected to FNOCT 8a to localize NO-dependent fluorescence in these roots; controls with an exogenous NO donor confirmed the presence and distribution of the probe in the roots. To confirm the NO-dependent fluorescence, roots were incubated with the three different NO scavengers cPTIO {2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-L-oxyl-3-oxide}, methylene blue and sodium diethyl dithiocarbamate (DCC) followed by incubation with FNOCT 8a. Methylene blue and DCC were able to completely quench NO-dependent fluorescence, cPTIO quenched partially. The roots were incubated in the presence of NaNO2 and NaNO3. which are substrates for nitrite:nitric oxide reductase (NI-NOR) and plasma membrane-bound nitrate reductase (PM-NR), respectively. The NO-dependent fluorescence was more or less same at the root tips upon treatment with NaNO2, while the overall fluorescence was reduced in the presence of NaNO3. Fluorescence from the living roots was visualized by inverted confocal laser scanning microscope (CLSM) using UV laser (excitation 360 nm and emission 408 nm). A specialized apparatus has been devised by the authors for analysis of intact roots as described in the methods section of this paper. Intact roots were chosen for microscopic observation rather than incised roots to avoid production of NO due to stress or physical injury. (C) 2012 Elsevier Masson SAS. All rights reserved.
语种	英语
公开日期	2015-06-08
内容类型	期刊论文
源URL	[http://ir.xtbg.org.cn/handle/353005/8834]
专题	西双版纳热带植物园_公共技术服务中心主要仪器相关文献_激光共聚焦显微镜
推荐引用方式 GB/T 7714	Vandana, S.. FNOCT as a fluorescent probe for in vivo localization of nitric oxide distribution in tobacco roots[J]. Plant Physiology and Biochemistry,2012,59:80-89.
APA	Vandana, S..(2012).FNOCT as a fluorescent probe for in vivo localization of nitric oxide distribution in tobacco roots.Plant Physiology and Biochemistry,59,80-89.
MLA	Vandana, S.."FNOCT as a fluorescent probe for in vivo localization of nitric oxide distribution in tobacco roots".Plant Physiology and Biochemistry 59(2012):80-89.