When applied under abiotic stress conditions, triacontanol (TRIA) is effective in regulating the physicochemical processes in plants through mechanisms of defence such as abscisic acid (ABA) signalling. However, TRIA’s role in relation to ABA and stomatal opening is unclear. Therefore, the objective of this study was to evaluate the effects of TRIA and ABA and their combinations on different variables related to stomatal regulation in Solanum lycopersicum, which is subjected to drought stress, and on the leaf epidermis. The negative effects of stress and responses triggered by ABA were reversed in plants treated with TRIA. TRIA increased stomatal conductance and photosynthetic activity in the early hours, and it was determined that TRIA produced larger stomata than did the other treatments. Moreover, the chloroplasts of plants treated with TRIA were significantly smaller and more numerous than those of the control, which could improve CO2 diffusion efficiency and may be related to the regulation of stomatal opening and photosynthesis. Finally, the abaxial epidermis tests reaffirmed the inhibitory effects of TRIA on ABA on stomatal opening. These results confirm the important role of TRIA in regulating various processes in plants and processes triggered by ABA, such as those related to stomatal regulation.