Soybeans (Glycine max) and grain amaranth (Amaranthus hypochondriacus) were grown at a range of temperatures, carbon dioxide concentrations and light conditions in controlled environment chambers, and the response of leaf conductance to water vapour to changes in humidity was then measured under a standard set of conditions. The sensitivity of conductance was analysed in terms of (i) the absolute sensitivity of conductance to changes in leaf to air water vapour pressure difference (LAVPD), (ii) the sensitivity of conductance relative to the absolute value of conductance, and (iii) the slope of the relationship between conductance and an index incorporating assimilation rate, carbon dioxide concentration and relative humidity. The sensitivity of conductance varied substantially with growth conditions for all three analyses in both species. The growth temperature of 25 °C increased the sensitivity of conductance by all three measures compared with growth at 20 or 30 °C in amaranth, with little difference between 25 and 30 °C in soybean. Growth at elevated carbon dioxide decreased sensitivity in amaranth by all three measures, and decreased the absolute but not the relative sensitivity in soybean. Growth at reduced photon flux density and growth at high stand density reduced sensitivity in amaranth by all three measures. In soybean, growth at high stand density reduced sensitivity by all three measures, but growth at low photon flux density increased the relative sensitivity. The sensitivity of leaf conductance to changes in humidity varied by a factor of two or more with growth environment by all measures of sensitivity in both the C3 and the C4 species.