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BREAKTHROUGH TECHNOLOGIES

Proton-Transfer-Reaction Mass Spectrometry as a New Tool for Real Time Analysis of Root-Secreted Volatile Organic Compounds in Arabidopsis¹

Aeronomy Laboratory, National Oceanic and Atmospheric Administration (NOAA), Boulder, Colorado 80305 (M.S., J.d.G., P.G., W.K., M.N.); Department of Molecular and Laser Physics, University of Nijmegen, Nijmegen, The Netherlands (M.S.); Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, Colorado 80309 (M.S., J.d.G., R.F.); Department of Horticulture and Landscape Architecture (H.P.B., J.M.V.), and Cell and Molecular Biology Program, Colorado State University, Fort Collins, Colorado 80523–1173 (J.M.V.); and Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado, 80309 (R.F.)

Plant roots release about 5% to 20% of all photosynthetically-fixed carbon, and as a result create a carbon-rich environment for numerous rhizosphere organisms, including plant pathogens and symbiotic microbes. Although some characterization of root exudates has been achieved, especially of secondary metabolites and proteins, much less is known about volatile organic compounds (VOCs) released by roots. In this communication, we describe a novel approach to exploring these rhizosphere VOCs and their induction by biotic stresses. The VOC formation of Arabidopsis roots was analyzed using proton-transfer-reaction mass spectrometry (PTR-MS), a new technology that allows rapid and real time analysis of most biogenic VOCs without preconcentration or chromatography. Our studies revealed that the major VOCs released and identified by both PTR-MS and gas chromatography-mass spectrometry were either simple metabolites, ethanol, acetaldehyde, acetic acid, ethyl acetate, 2-butanone, 2,3,-butanedione, and acetone, or the monoterpene, 1,8-cineole. Some VOCs were found to be produced constitutively regardless of the treatment; other VOCs were induced specifically as a result of different compatible and noncompatible interactions between microbes and insects and Arabidopsis roots. Compatible interactions of Pseudomonas syringae DC3000 and Diuraphis noxia with Arabidopsis roots resulted in the rapid release of 1,8-cineole, a monoterpene that has not been previously reported in Arabidopsis. Mechanical injuries to Arabidopsis roots did not produce 1,8-cineole nor any C₆ wound-VOCs; compatible interactions between Arabidopsis roots and Diuraphis noxia did not produce any wound compounds. This suggests that Arabidopsis roots respond to wounding differently from above-ground plant organs. Trials with incompatible interactions did not reveal a set of compounds that was significantly different compared to the noninfected roots. The PTR-MS method may open the way for functional root VOC analysis that will complement genomic investigations in Arabidopsis.

² These authors contributed equally to the paper.

www.plantphysiol.org/cgi/doi/10.1104/pp.104.038703.

^* Corresponding author; e-mail jvivanco{at}lamar.colostate.edu; fax 970–491–7745.

Received January 5, 2004; returned for revision February 5, 2004; accepted February 5, 2004.

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