Results: We generated 43 Gb of short Illumina reads and 9 Gb of long Nanopore reads, representing approximate genome coverage of 54× and 11×, respectively, based on the range of estimated k-mer-predicted genome sizes of between 791 and 967 Mbp. The final assembled genome is contained in 6404 scaffolds with an accumulated length of 880 Mb (96.3% BUSCO-calculated genome completeness). Compared with the Illumina-only assembly, the hybrid approach generated 94% fewer scaffolds with an 18-fold increase in N50 length (401 kb) and increased the genome completeness by an additional 16%. A total of 27 240 high-quality protein-coding genes were predicted from the clown anemonefish, 26 211 (96%) of which were annotated functionally with information from either sequence homology or protein signature searches.
Conclusions: We present the first genome of any anemonefish and demonstrate the value of low coverage (∼11×) long Nanopore read sequencing in improving both genome assembly contiguity and completeness. The near-complete assembly of the A. ocellaris genome will be an invaluable molecular resource for supporting a range of genetic, genomic, and phylogenetic studies specifically for clownfish and more generally for other related fish species of the family Pomacentridae.
METHODS: Fresh specimens of chondracanthids were collected from the buccal cavity of two species of deep-sea fishes (fish hosts were frozen), Chaunax abei Le Danois, 1978 (Lophiiformes: Chaunacidae) and Setarches longimanus (Alcock, 1894) (Perciformes: Setarchidae), caught at a depth of 212 m in Suruga Bay, Japan (34° 37'48.87″ N, 138° 43'2.958″ E). Both the species are described and illustrated based on ovigerous females.
RESULTS: The genus Avatar gen. nov. can readily be distinguished from all other chondracanthid genera by the following combination of features: cephalothorax slightly wider than long with anterior pair of large and posterior pair of small lateral lobes, and two pairs of ventro-lateral processes; the very posteriormost part of the first pedigerous somite contributes to the neck; cylindrical trunk with two pairs of blunt proximal fusiform processes; antennule with small knob terminally; antenna bearing distal endopodal segment; labrum protruding ventrally; two pairs of biramous legs each with 2-segmented rami. Kokeshioides gen. nov. has the following combinations of features that distinguish it from other chondracanthid genera: body flattened, without lateral processes; cephalothorax much wider than long, with paired anterolateral and posterolateral lobes, folded ventrally; the very posteriormost part of the first pedigerous somite contributes to the neck; mandible elongate; legs unique, heavily sclerotized, represented by two pairs of acutely pointed processes.
CONCLUSION: With the addition of two new genera presently reported, the family Chondracanthidae currently includes 52 valid genera. Among the described genera Avatar gen. nov. seems to be very primitive, while Kokeshioides gen. nov. is highly advanced. The deduced evolutionary history of chondracanthid genera is also discussed.