High relative air humidity (RH ≥ 85%) during growth disturbs stomatal functioning, resulting in excessive water loss in conditions of high evaporative demand. We investigated the expression of nine abscisic acid (ABA)-related genes (involved in ABA biosynthesis, oxidation and conjugation) and two non-ABA related genes (involved in the water stress response) aiming to better understand the mechanisms underlying contrasting stomatal functioning in plants grown at high RH. Four rose genotypes with contrasting sensitivity to high RH (one sensitive, one tolerant and two intermediate) were grown at moderate (62 ± 3%) or high (89 ± 4%) RH. The sensitive genotype grown at high RH showed a significantly higher stomatal conductance (gs) and water loss in response to closing stimuli as compared to the other genotypes. Moreover, high RH reduced the leaf ABA concentration and its metabolites to a greater extent in the sensitive genotype as compared to the tolerant one. The large majority of the studied genes had a relevant role on stomatal functioning (NCED1, UGT75B2, BG2, OST1, ABF3 and Rh-APX) while two others showed a minor contribution (CYP707A3 and BG1) and AAO3, CYP707A1 and DREB1B did not contribute to the tolerance trait. These results show that multiple genes form a highly complex regulatory network acting together towards the genotypic tolerance to high RH.