Alfalfa (Medicago sativa L,) production under irrigated and rainfed conditions may benefit from improvements in water-use efficiency (WUE), the amount of forage and root biomass produced per unit of water transpired. If benefits from improved WUE are to be realized, correlations between important agronomic traits and key physiological traits associated with WUE must be determined. This study characterized variation for dry matter yield, forage maturity, leaf-to-stem ratio (LSR), carbon isotope discrimination (Delta), canopy temperature, ash content, and specific leaf mass (SLM) in alfalfa. Associations between traits were also determined. Nine alfalfa germplasms representing eight of the nine historical genetic diversity groups, and a very fall-dormant (VFD) population, were established in seeded, irrigated plots for 2 yr near Las Cruces, NM. Significant variation (P less than or equal to 0.10) was detected for all traits and was greatest for Delta and maturity, intermediate for yield, canopy temperature, ash content, and LSR, and least for SLM. The African, Peruvian, and Indian germplasms exhibited a higher Delta than either the Turkistan, VFD, M. varia Martyn., or Ladak germplasms, Carbon isotope discrimination was positively correlated with forage yield (r = 0.64; P less than or equal to 0.10; n = 9) and forage maturity (r = 0.66; P less than or equal to 0.05; n = 9), No association was detected between Delta and either canopy temperature, ash content, SLM, or LSR. The results indicate that differences in stomatal conductance or photosynthetic capacity exist among the nine populations, and that germplasms with low Delta tended to have slower growth and development rates under irrigated conditions. Neither canopy temperature, ash content, nor SLM provided suitable alternate measurements of Delta among the nine alfalfa germplasms.