Tropospheric ozone (O3) is a global air pollutant that adversely affects plant growth. Whereas the impacts of O3 have previously been examined for some tropical commodity crops, no information is available for the pantropical crop, banana (Musa spp.). To address this, we exposed Australia’s major banana cultivar, Williams, to a range of [O3] in open top chambers. In addition, we examined 46 diverse Musa lines growing in a common garden for variation in three traits that are hypothesised to shape responses to O3: (1) leaf mass per area; (2) intrinsic water use efficiency; and (3) total antioxidant capacity. We show that O3 exposure had a significant effect on the biomass of cv. Williams, with significant reductions in both pseudostem and sucker biomass with increasing [O3]. This was accompanied by a significant increase in total antioxidant capacity and phenolic concentrations in older, but not younger, leaves, indicating the importance of cumulative O3 exposure. Using the observed trait diversity, we projected O3 tolerance among the 46 Musa lines growing in the common garden. Of these, cv. Williams ranked as one of the most O3-tolerant cultivars. This suggests that other genetic lines could be even more susceptible, with implications for banana production and food security throughout the tropics.