The function in plants of the non-protein amino acid, gamma-aminobutyric acid (GABA) is poorly understood. In this study, we show that GABA down-regulates the expression of a large sub-set of 14-3-3 gene family members in Arabidopsis thaliana seedlings in a calcium, ethylene and abscisic acid-dependent manner. Gene expression is not affected when seedlings are supplied with glutamate, a precursor of GABA. The repression of 14-3-3 gene expression by GABA is dependent on functional ethylene and abscisic acid signalling pathways, since the response is lost in the etr1-1, abi1-1 and abi2-1 mutants. Calcium measurements show that in contrast to glutamate, GABA does not elicit a cytoplasmic calcium elevation, suggesting that the GABA response is unlikely to be mediated by glutamate receptors, as has been suggested previously. We suggest that in addition to its role as a stress-related metabolite, GABA may regulate gene expression in Arabidopsis, including members of the 14-3-3 gene family.