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Background   Heliconius  butterflies possess a remarkable diversity of phenotypes, physiologies, and behaviors that has long distinguished this genus as a focal taxon in ecological and evolutionary research. Recently  Heliconius  has also emerged as a model system for using genomic methods to investigate the causes and consequences of biological diversity. One notable aspect of  Heliconius  diversity is a dichotomy in mating systems which provides an unusual opportunity to investigate the relationship between sexual selection and the evolution of reproductive proteins. As a first step in pursuing this research, we report the generation and analysis of expressed sequence tags (ESTs) from the male accessory gland of  H. erato  and  H. melpomene , species representative of the two mating systems present in the genus  Heliconius .    Results  We successfully sequenced 933 ESTs clustering into 371 unigenes from  H. erato  and 1033 ESTs clustering into 340 unigenes from  H. melpomene . Results from the two species were very similar. Approximately a third of the unigenes showed no significant BLAST similarity (E-value <10 -5 ) to sequences in GenBank's non-redundant databases, indicating that a large proportion of novel genes are expressed in  Heliconius  male accessory glands. In both species only a third of accessory gland unigenes were also found among genes expressed in wing tissue. About 25% of unigenes from both species encoded secreted proteins. This includes three groups of highly abundant unigenes encoding repetitive proteins considered to be candidate seminal fluid proteins; proteins encoded by one of these groups were detected in  H. erato  spermatophores.    Conclusion  This collection of ESTs will serve as the foundation for the future identification and evolutionary analysis of male reproductive proteins in  Heliconius  butterflies. These data also represent a significant advance in the rapidly growing collection of genomic resources available in  Heliconius  butterflies. As such, they substantially enhance this taxon as a model system for investigating questions of ecological, phenotypic, and genomic diversity.

EST analysis of male accessory glands from Heliconius butterflies with divergent mating systems