Sulfur is an essential macroelement in plant and animal nutrition. Plants assimilate inorganic sulfate into two sulfur-containing amino acids, cysteine and methionine. Low supply of sulfate leads to decreased sulfur pools within plant tissues. As sulfurrelated metabolites represent an integral part of plant metabolism with multiple interactions, sulfur deficiency stress induces a number of adaptive responses, which must be coordinated. To reveal the coordinating network of adaptations to sulfur deficiency, metabolite profiling of Arabidopsis has been undertaken. Gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry techniques revealed the response patterns of 6,023 peaks of nonredundant ion traces and relative concentration levels of 134 nonredundant compounds of known chemical structure. Here, we provide a catalogue of the detected metabolic changes and reconstruct the coordinating network of their mutual influences. The observed decrease in biomass, as well as in levels of proteins, chlorophylls, and total RNA, gives evidence for a general reduction of metabolic activity under conditions of depleted sulfur supply. This is achieved by a systemic adjustment of metabolism involving the major metabolic pathways. Sulfur/carbon/nitrogen are partitioned by accumulation of metabolites along the pathway O-acetylserine to serine to glycine, and are further channeled together with the nitrogen-rich compound glutamine into allantoin. Mutual influences between sulfur assimilation, nitrogen imbalance, lipid breakdown, purine metabolism, and enhanced photorespiration associated with sulfur-deficiency stress are revealed in this study. These responses may be assembled into a global scheme of metabolic regulation induced by sulfur nutritional stress, which optimizes resources for seed production.For living organisms with a complex hierarchical organization, such as plants, the necessity for close coordination of various elements requires systemic organization at the level of the whole organism. Additional complexity is imposed on the system through environmental variability. Due to the inability to escape unfavorable environmental conditions, plants have evolved complex mechanisms to sense and transmit external signals to the internal decision points to trigger the adaptive response program for homeostatic maintenance. Such adaptive programs are accomplished at multiple organizational levels, e.g. gene and enzymatic activities, being finally manifested in altered metabolite concentrations. This study describes the metabolic component of the whole-system response to sulfur deficiency, as an environmental perturbation, with special emphasis on the mechanisms of the response coordination.Despite the difficulties in measuring metabolites due to their dynamic behavior and complex chemistry, new methods allow profiling of low molecular weight compounds, with gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) being the most robust (Stitt and Fernie, 2003). In th...