Fig. 4. Male-specific compounds regulate intraspecific sexual behaviors and interspecific sexual isolation.

a Top left: Names of the compounds that are exclusively produced by males of 54 species (left below) and detected by conspecific females through at1 or/and at4 (right below). These compounds were used for the behavioral experiments in Fig. 4b–d. b Top: Schematic of a mating arena where females of each species had the choice to mate with two conspecific males perfumed with their olfactory-detected male-specific compound (indicated, in Fig. 4a, on the left side of the horizontal dashed line) or solvent (dichloromethane, DCM). For consistency of perfuming and correspondence to biologically relevant amounts see “Methods”. Below: bar plots represent the percentages of copulation success of the rival males. Results from females that were only courted by one male were excluded. In this and other panels, filled bars indicate a significant difference between the tested groups; ns P > 0.05; *P < 0.05; **P < 0.01; ***P < 0.001, chi-square test. Number of replicates are stated on the left side on the bar plots. See Supplementary Fig. 4A for details regarding the differences and similarities of sexual behaviors among the 54 species. Note that in 11 instances, females displayed a preference to copulate with the male-specific compound-perfumed males over the control ones, while 6 compounds resulted in avoidance, and 29 turned out to be neutral. See Supplementary Data 7 for raw data and statistical analyses. See Supplementary Fig. 4B for the effect of perfuming on the males’ courtship behavior. Drawings made by Mohammed A. Khallaf. c Top: Competition courtship arenas where a male of each species had the choice to court two decapitated conspecific females perfumed with the male-transferred compound. Note that we tested only transferred compounds (green). Below: bar plots represent the percentage of the first copulation attempts towards perfumed and control females. Results from males that only courted one female were excluded; see Methods. Note that 15 compounds inhibited courtship, 1 compound increased courtship and 16 compounds turned out to be neutral. Drawings made by Mohammed A. Khallaf. d Top: Schematic of a mating arena where a female of each species had the choice to mate with two conspecific males perfumed with olfactory-detected heterospecific cVA or solvent (DCM). Note that we only tested the species that do not produce but still detect cVA. Below: bar plots represent the percentages of copulation success of the rival males. Results from females that were only courted by one male were excluded. Drawings made by Mohammed A. Khallaf.