Many species display local variations in pre-mating signals and in mating preferences. This may lead to discrimination against potential foreign mates that may ultimately lead to reproductive isolation. However, the extent to which population differentiation in mating cues affects the species recognition has received little empirical support. Here we investigate the consequence of geographic differentiation in male reproductive traits on female preferences to these traits in Bombus terrestris. We characterise (1)the geographic differentiation in male cephalic labial gland secretions (CLGS), a key trait for mate attraction and (2) the preference of virgin females to the CLGS of different subspecies. Our results show geographic CLGS differences parallel with divergences in female preferences for these secretions. This geographic CLGS differentiation in males, along with female preference for sympatric males could lead to or reflect a premating isolation among subspecies.