Do pollen feeding, pupal‐mating and larval gregariousness have a single origin in Heliconius butterflies? Inferences from multilocus DNA sequence data

Phylogenetic information is useful in understanding the evolutionary history of adaptive traits. Here, we present a well‐resolved phylogenetic hypothesis for Heliconius butterflies and related genera. We use this tree to investigate the evolution of three traits, pollen feeding, pupal‐mating behaviour and larval gregariousness. Phylogenetic relationships among 60 Heliconiina species (86% of the subtribe) were inferred from partial DNA sequences of the mitochondrial genes cytochrome oxidase I , cytochrome oxidase II and 16S rRNA, and fragments of the nuclear genes elongation factor‐1α , apterous , decapentaplegic and wingless (3834 bp in total). The results corroborate previous hypotheses based on sequence data in showing that Heliconius is paraphyletic, with Laparus doris and Neruda falling within the genus, demonstrating a single origin for pollen feeding but with a loss of the trait in Neruda . However, different genes are not congruent in their placement of Neruda ; therefore, monophyly of the pollen feeding species cannot be ruled out. There is also a highly supported monophyletic ‘pupal‐mating clade’ suggesting that pupal mating behaviour evolved only once in the Heliconiina. Additionally, we observed at least three independent origins for larval gregariousness from a solitary ancestor, showing that gregarious larval behaviour arose after warning coloration. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 221–239.