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

Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Bibliographische Detailangaben
Veröffentlicht in:Plant physiology and biochemistry : PPB. - 1991. - 59(2012) vom: 30. Okt., Seite 80-9
1. Verfasser: Vandana, Shweta (VerfasserIn)
Weitere Verfasser: Sustmann, Reiner, Rauen, Ursula, Stöhr, Christine
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2012
Zugriff auf das übergeordnete Werk:Plant physiology and biochemistry : PPB
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Fluorescent Dyes Membrane Proteins Nitrates Plant Proteins Nitric Oxide 31C4KY9ESH sodium nitrate 8M4L3H2ZVZ mehr... Oxidoreductases EC 1.- nitric-oxide reductase EC 1.7.2.5 Nitrate Reductase EC 1.7.99.4 Sodium Nitrite M0KG633D4F
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520 |a Copyright © 2012 Elsevier Masson SAS. All rights reserved. 
520 |a 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 NaNO. 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 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 7 |a Fluorescent Dyes  |2 NLM 
650 7 |a Membrane Proteins  |2 NLM 
650 7 |a Nitrates  |2 NLM 
650 7 |a Plant Proteins  |2 NLM 
650 7 |a Nitric Oxide  |2 NLM 
650 7 |a 31C4KY9ESH  |2 NLM 
650 7 |a sodium nitrate  |2 NLM 
650 7 |a 8M4L3H2ZVZ  |2 NLM 
650 7 |a Oxidoreductases  |2 NLM 
650 7 |a EC 1.-  |2 NLM 
650 7 |a nitric-oxide reductase  |2 NLM 
650 7 |a EC 1.7.2.5  |2 NLM 
650 7 |a Nitrate Reductase  |2 NLM 
650 7 |a EC 1.7.99.4  |2 NLM 
650 7 |a Sodium Nitrite  |2 NLM 
650 7 |a M0KG633D4F  |2 NLM 
700 1 |a Sustmann, Reiner  |e verfasserin  |4 aut 
700 1 |a Rauen, Ursula  |e verfasserin  |4 aut 
700 1 |a Stöhr, Christine  |e verfasserin  |4 aut 
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773 1 8 |g volume:59  |g year:2012  |g day:30  |g month:10  |g pages:80-9 
856 4 0 |u http://dx.doi.org/10.1016/j.plaphy.2012.01.004  |3 Volltext 
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