Real-time electrochemical detection of extracellular nitric oxide in tobacco cells exposed to cryptogein, an elicitor of defence responses

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Abstract

It was previously reported that cryptogein, an elicitor of defence responses, induces an intracellular production of nitric oxide (NO) in tobacco. Here, the possibility was explored that cryptogein might also trigger an increase of NO extracellular content through two distinct approaches, an indirect method using the NO probe 4,5-diaminofluorescein (DAF-2) and an electrochemical method involving a chemically modified microelectrode probing free NO in biological media. While the chemical nature of DAF-2-reactive compound(s) is still uncertain, the electrochemical modified microelectrodes provide real-time evidence that cryptogein induces an increase of extracellular NO. Direct measurement of free extracellular NO might offer important new insights into its role in plants challenged by biotic stresses.

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Most cited references 47

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ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis.

Jo Bright, Radhika Desikan, John Hancock … (2006)

Nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) are key signalling molecules produced in response to various stimuli and involved in a diverse range of plant signal transduction processes. Nitric oxide and H(2)O(2) have been identified as essential components of the complex signalling network inducing stomatal closure in response to the phytohormone abscisic acid (ABA). A close inter-relationship exists between ABA and the spatial and temporal production and action of both NO and H(2)O(2) in guard cells. This study shows that, in Arabidopsis thaliana guard cells, ABA-mediated NO generation is in fact dependent on ABA-induced H(2)O(2) production. Stomatal closure induced by H(2)O(2) is inhibited by the removal of NO with NO scavenger, and both ABA and H(2)O(2) stimulate guard cell NO synthesis. Conversely, NO-induced stomatal closure does not require H(2)O(2) synthesis nor does NO treatment induce H(2)O(2) production in guard cells. Tungstate inhibition of the NO-generating enzyme nitrate reductase (NR) attenuates NO production in response to nitrite in vitro and in response to H(2)O(2) and ABA in vivo. Genetic data demonstrate that NR is the major source of NO in guard cells in response to ABA-mediated H(2)O(2) synthesis. In the NR double mutant nia1, nia2 both ABA and H(2)O(2) fail to induce NO production or stomatal closure, but in the nitric oxide synthase deficient Atnos1 mutant, responses to H(2)O(2) are not impaired. Importantly, we show that in the NADPH oxidase deficient double mutant atrbohD/F, NO synthesis and stomatal closure to ABA are severely reduced, indicating that endogenous H(2)O(2) production induced by ABA is required for NO synthesis. In summary, our physiological and genetic data demonstrate a strong inter-relationship between ABA, endogenous H(2)O(2) and NO-induced stomatal closure.

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NO news is good news for plants.

Massimo Delledonne (2005)

The organization of redox signaling and the use of nitric oxide (NO) to transmit information, modulate biological processes or create cellular damage are highly complex. Recent reports provide an exceptional picture of NO production, of the regulation of NO bioactivity through detoxification reactions and of biochemical events by which NO transduces signals into cellular responses, in particular during disease resistance. Furthermore, other exciting reports on NO function in germination, growth and reproduction support the view that NO is a 'do it all' molecule that plays a crucial role during the entire lifespan of the plant.

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Nitric oxide is involved in growth regulation and re-orientation of pollen tubes.

D Porterfield, Flaviane Prado, José A Feijó (2004)

Nitric oxide (NO) controls diverse functions in many cells and organs of animals. It is also produced in plants and has a variety of effects, but little is known about their underlying mechanisms. In the present study, we have discovered a role for NO in the regulation of pollen tube growth, a fast tip-growing cellular system. Pollen tubes must be precisely oriented inside the anatomically complex female ovary in order to deliver sperm. We hypothesized that NO could play a role in this guidance and tested this hypothesis by challenging the growth of pollen tubes with an external NO point source. When a critical concentration was sensed, the growth rate was reduced and the growth axis underwent a subsequent sharp reorientation, after which normal growth was attained. This response was abrogated in the presence of the NO scavenger CPTIO and affected by drugs interfering in the cGMP signaling pathway. The sensitivity threshold of the response was significantly augmented by sildenafil citrate (SC), an inhibitor of cGMP-specific phosphodiesterases in animals. NO distribution inside pollen tubes was investigated using DAF2-DA and was shown to occur mostly in peroxisomes. Peroxisomes are normally excluded from the tip of pollen tubes and little if any NO is found in the cytosol of that region. Our data indicate that the rate and orientation of pollen tube growth is regulated by NO levels at the pollen tube tip and suggest that this NO function is mediated by cGMP.

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Author and article information

Journal

Journal ID (nlm-ta): J Exp Bot

Journal ID (hwp): jexbot

Journal ID (publisher-id): exbotj

Title: Journal of Experimental Botany

Publisher: Oxford University Press

ISSN (Print): 0022-0957

ISSN (Electronic): 1460-2431

Publication date (Print): September 2008

Publication date (Electronic): 24 July 2008

Publication date PMC-release: 24 July 2008

Volume: 59

Issue: 12

Pages: 3407-3414

Affiliations

[1 ]UMR INRA 1088/CNRS 5184/Université de Bourgogne, Plante-Microbe-Environnement, BP 86510, 17 rue Sully, 21000 Dijon, France

[2 ]CNRS, UMR 8151, Ecole Nationale Supérieure de Chimie de Paris, INSERM, U640, University Paris Descartes. ENSCP, 11 rue Pierre et Marie Curie, 75231 Paris cedex 05, France

Author notes

[† ]To whom correspondence should be addressed. E-mail: fethi-bedioui@ 123456enscp.fr

[† ]To whom correspondence should be addressed. E-mail: wendehen@ 123456dijon.inra.fr

[*]

These authors contributed equally to this work.

Article

DOI: 10.1093/jxb/ern189

PMC ID: 2529233

PubMed ID: 18653691

SO-VID: 247a4e19-849a-4d25-a567-bb328c1f0064

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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

Real-time electrochemical detection of extracellular nitric oxide in tobacco cells exposed to cryptogein, an elicitor of defence responses

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Most cited references 47

ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis.

NO news is good news for plants.

Nitric oxide is involved in growth regulation and re-orientation of pollen tubes.

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