Variation in floral phenotype (color, depth, nectar) suggests incipient specialization for bee or hawkmoth pollination across the geographic distribution of Echinopsis ancistrophora, with flower depth ranging from 4.5 to 24 cm. We used chemical and behavioral analyses to test whether fragrance has evolved in concert with morphology in these Andean cacti. Floral scent (145 total compounds) was collected using dynamic headspace methods and analyzed with gas chromatography–mass spectrometry, revealing subspecies-specific odors dominated by sesquiterpenes in E. ssp. ancistrophora and arachnacantha and fatty acid derivatives or aromatics in E. ssp. cardenasiana and pojoensis. Compounds indicative of sphingophily were not consistently found in moth-pollinated plants, and total scent emissions were significantly lower in populations with nocturnal anthesis. In wind tunnel assays, Manduca sexta moths were attracted to scent of ssp. ancistrophora from both bee and hawkmoth-pollinated populations, but not to scent of ssp. cardenasiana. However, hawkmoths were most attracted to the methyl benzoate-dominated scent of a distant relative, Echinopsis mirabilis. Thus, hawkmoth-pollinated descendants of the E. ancistrophora lineage may be phylogenetically constrained to emit weak, sesquiterpene-dominated fragrances that are not optimally attractive to hawkmoths, or floral scent may be under stronger selection by destructive flower visitors.