We studied the response of stomatal conductance at leaf (gS) and canopy (GS) scales to increasing vapor pressure deficit (D) in mature Pinus palustris Mill. (longleaf pine) growing in a sandhill habitat in the coastal plain of the southeastern USA. Specifically, we determined if variation in the stomatal response to D was related to variation in hydraulic conductance along the soil-to-leaf pathway (KL) over the course of a growing season. Reductions in KL were associated with a severe growing season drought that significantly reduced soil water content (theta) in the upper 90-cm soil profile. Although KL recovered partially following the drought, it never reached pre-drought values. Stomatal sensitivity to D was well correlated with maximum gS at low D at both leaf and canopy scales, and KL appeared to influence this response by controlling maximum gS. Our results are consistent with the hypothesis that stomatal response to D occurs to regulate minimum leaf water potential, and that the sensitivity of this response is related to changes in whole-plant hydraulics.