The involvement of metallothioneins (MTs) in response to plant water stress and recovery was assessed by analyzing gene expression in leaves and in the cambial zone of white poplar. One-year-old plants were submitted to two different watering regimes: irrigation was withheld for 9d and then resumed until day 17, or soil moisture was maintained to field capacity by irrigation during the experiment. Changes in leaves and stem water relations, gas exchange and CO(2) assimilation were recorded. The expression profiles of MT genes were analyzed in developing leaves and the cambial zone at maximum stress levels and after recovery and compared with the watered controls. Whole-plant water relations were significantly affected by water deprivation, though a complete recovery of plant water status was reached after resumption of watering. Withholding irrigation resulted in a significant decrease of leaf turgor potential and relative water content without a significant increase of the osmotic potential at full turgor. Similarly, stem water content decreased, leading to a marked increase of stem shrinkage, confirming that mild water stress affected primarily tissue water status. Following water depletion, the transcript analysis of MT genes revealed increased expression of type 3a and 3b MT genes in cambial tissues, and particularly in leaves. After water resumption, transcription decreased, suggesting that the changes in gene expression were related to water deficit. The results indicate that in leaves and, for the first time, in the cambial zone, type 3 MTs respond in a specific manner to changes in water status. These results are consistent with the regulatory cis-elements present in the 5' flanking region of type 3 MT genes.