Sequencing and Annotation of Transcriptomes from Two Anemonefish Species

Anemonefishes (clownfishes) are popular among marine aquarium hobbyists, largely because of their bright colors and their fame from children’s movies and books.  For this reason, they are the most widely bred species in marine ornamental aquaculture, and several fancy varieties have been developed.  In addition, anemonefish exhibit many remarkable adaptations that have yet to be deciphered, including mutualistic interactions with sea anemones and serial hermaphroditism.  As an initial step to develop genomic tools, we have sequenced the transcriptomes of two anemonefish species, the false clownfish (Amphiprion ocellaris) and the two-band anemonefish (A. bicinctus), using a combination of Roche/454 and Illumina sequencing.  An initial de novo assembly using CLC Genomic Workbench produced 28,035 contigs for A. ocellaris and 14,124 for A. bicinctus.  These contigs and accompanying singletons were annotated using the KEGG Automatic Annotation Server (KAAS) and by BLASTx comparisons to the SwissProt database and to the proteomes of Oreochromis niloticus, Takifugu rubripes, Oryzias latipes, and Danio rerio.  This work will serve as a reference transcriptome for future investigations, and has provided the foundation for undergraduate classroom research experiences investigating gene expression in anemonefish.