A major hypothesis for the evolution of chemical signals is that pheromones arise from non-communicative precursor compounds. However, data supporting this hypothesis are rare, primarily because the original functions of the antecedent compounds often have been lost. A notable exception, however, is the parasitoid wasp species Leptopilina heterotoma, whose compound (−)-iridomyrmecin is used as a defensive secretion, a cue for females to avoid competition with con- and hetero-specific females, and as the primary component of the females’ sex pheromone. To better understand the evolution of sex pheromones from defensive compounds, we examined the chemical ecology of L. pacifica, the sister species of L. heterotoma. Here, we show that L. pacifica also produces a defensive secretion containing a species-specific mixture of mostly iridoid compounds. However, the composition of the secretion is more complex than in L. heterotoma, and iridomyrmecin is only a minor component. Moreover, in contrast to L. heterotoma, conspecific female competitors were not avoided by female subjects, and a role of the iridoids in the female sex pheromone of L. pacifica can be excluded, as only the females’ cuticular hydrocarbons (CHCs) resulted in the elicitation of courtship by males. Although closely related, the two sister species show substantial differences in the use of the defensive secretion for communicative purposes. Variation in pheromone usage in this genus still presents a conundrum, highlighting the need for additional studies to understand the selective forces shaping the evolution of pheromone composition.
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