The use of vapor phase extraction in metabolic profiling of phytohormones and other metabolites.

Through complex networks of signaling interactions, phytohormones regulate growth, development, reproduction and responses to biotic and abiotic stress. Comprehensive metabolomic approaches, seeking to quantify changes in vast numbers of plant metabolites, may ultimately clarify these complex signaling interactions and consequently explain pleiotropic effects on plant metabolism. Synergistic and antagonistic phytohormone signaling interactions, referred to as crosstalk, are often considered at the level of transduction without proper consideration of synthesis or accumulation of phytohormones because of the limitation and difficulty in quantifying numerous signals. Significant progress has recently been made in the expansion of metabolic profiling and analysis of multiple phytohormones [Birkemeyer et al. (J. Chromatogr. A, 2003, 993, 89); Chiwocha et al. (Plant J., 2003, 35, 405); Müller et al. (Planta, 2002, 216, 44); Schmelz et al. (Proc. Natl Acad. Sci. USA, 2003, 100, 10552)]. We recently presented a novel metabolic profiling approach to the analysis of acidic phytohormones and other metabolites based on a simplistic preparation scheme and analysis by chemical ionization-gas chromatography/mass spectrometry. We now provide a detailed description of this vapor phase extraction technique and use pathogen infection of Arabidopsis with Pseudomonas syringae DC3000 to illustrate metabolic changes in salicylic acid, cinnamic acid, jasmonic acid, indole-3-acetic acid, abscisic acid, unsaturated C(18) fatty acids, 12-oxo-phytodienoic acid, and phytotoxin coronatine. Directions for further method expansion are provided and include issues of recovery, derivatization, range of accessible analytes, optimization, reproducibility and future directions.