Limited water availability is a major constraint to canola (Brassica napus L.) yield in the Mediterranean-type climate region. Selecting and breeding for genotypes with conservative water use characteristics is a promising strategy to improve yield in this environment. Three experiments were conducted to investigate transpiration responses (TR) to vapour pressure deficit (VPD) and progressive soil drying with 8-20 canola genotypes. We used the linear-plateau model to describe TR to elevated VPD and decreased fraction of transpirable soil water (FTSW) and identified the VPD and FTSW thresholds for plant to limit its transpiration. Canola genotypes showed significant variations in both VPD and FTSW thresholds. The genotypes with conservative water use reduced TR at a lower VPD threshold and decreased TR at a higher FTSW threshold than the profligate ones. We found that the conservative genotypes had low VPD and high FTSW thresholds while the profligate ones had high VPD and low FTSW thresholds. This conservative and profligate water use characteristics were consistent during both vegetative and reproductive stages. Furthermore, the relative yield of genotypes under drought conditions was positively related to the FTSW thresholds during the reproductive stage, indicating the better relative yield performance of conservative genotypes in water-limited farming system. We conclude that canola genotypes with lower VPD and higher FTSW thresholds could conserve water and defer water use for reproductive growth while the profligate genotypes can be deployed to take advantage of high rainfall in the high rainfall zone of southern Australia.