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Abstract To produce pitaya (Hylocereus spp.), it is essential that pollination occur, either artificially or by pollinating agents. This study evaluated the viability of self-pollination, cross-pollination, and natural pollination, as well as pollen viability, stigma receptivity and ideal pollination window in pitaya flowers (Hylocereus spp.). An experiment was conducted with a randomized block design consisting of four treatments: T1: manual self-pollination; T2: nocturnal open pollination; T3: diurnal open pollination; and T4: manual cross-pollination - red-fleshed pitaya [ Hylocereus polyrhizus] pollen placed on white-fleshed pitaya [ Hylocereus undatus] stigma. The experiment had four replicates, with two plants per replicate (two flowers per plant), totaling 16 flowers per treatment. The analyzed variables were fruit weight, length, diameter, titratable acidity, pH, soluble solids, and SS/TA ratio; the germination percentage of pollen grains; and the receptivity of stigmas. Artificial pollination (self- and cross-pollination) of white-fleshed pitaya (Hylocereus undatus) is viable, resulting in larger fruits than natural pollination, with cross-pollination being the most recommended. Throughout the floral opening period, the pitaya flower (Hylocereus spp.) has receptive stigma capable of receiving pollen. The flowers exhibit the highest pollen germination rate at 7 p.m, which is the ideal pollination window for pitaya.