Climate change is increasing drought events and decreasing water availability. Tomato is commonly transplanted to an open field after seedling production in a nursery, requiring large volumes of water. Arbuscular mycorrhizal (AM) fungi help plants cope with drought stress; however, their effects depend on plant genotype and environmental conditions. In this study, we assessed the interactions among different tomato seedling genotypes and two AM fungi, Funneliformis mosseae and Rhizophagus intraradices, under two water regimes, full and reduced. Our results showed that F. mosseae was more effective than R. intraradices in the mitigation of drought stress both in old and modern genotypes. However, seedlings inoculated with R. intraradices recorded the highest values of leaf area. ‘Pearson’ and ‘Everton’ genotypes inoculated with F. mosseae recorded the highest values of root, leaf, and total dry weights under reduced and full irrigation regimes, respectively. In addition, ‘Pearson’ and ‘H3402’ genotypes inoculated with F. mosseae under a reduced irrigation regime displayed high values of water use efficiency. Our results highlight the importance of using AM fungi to mitigate drought stress in nursery production of tomato seedlings. However, the development of ad hoc AM fungal formulations, which consider genotype x AM fungi interactions, is fundamental for achieving the best agronomic performances.