Woody plants can be induced to cold-acclimate by exposure to sublethal low temperatures, but only after the onset of vegetative maturity. We monitored seven woody plant taxa, at monthly intervals, to determine the date of vegetative maturity, freeze-killing temperature, cell membrane electrolyte leakage, and the quantity and diversity of endogenous oligosaccharides. The freeze-killing temperature changed from -5 to -7C before vegetative maturity to -15 to -20C after vegetative maturity. There was a 10-fold increase in raffinose and about a 3-fold increase in endogenous stachyose in samples that were cold-acclimated under controlled conditions. In field samples, endogenous raffinose increased from <0.02% in August to 2% to 11% in cortical stem tissues of all cold-acclimated taxa. The tetrasaccharide stachyose increased from <0.02% to 0.25% to 2.5% for similar comparisons. None of the other sugars or polyols showed similar, consistent patterns during the onset of cold acclimation. In response to low temperature, raffinose family oligosaccharides (RFOs) have previously been shown to increase substantially in cabbage, soybean, kidney bean, and Chlorella. RFOs also possess high water-binding characteristics and tend to enhance aqueous glass transitions. Accordingly, we hypothesize that the endogenous production of these oligosaccharides may play an important role in metabolic events associated with cryoprotection of critical cellular functions during low-temperature stress.