title: Next-generation sequencing, phylogenetic signal and comparative mitogenomic analyses in Metacrangonyctidae (Amphipoda: Crustacea).
abstract: BACKGROUND
  Comparative mitochondrial genomic analyses are rare among crustaceans below the family or genus level. The obliged subterranean crustacean amphipods of the family Metacrangonyctidae, found from the Hispaniola (Antilles) to the Middle East, including the Canary Islands and the peri-Mediterranean region, have an evolutionary history and peculiar biogeography that can respond to Tethyan vicariance. Indeed, recent phylogenetic analysis using all protein-coding mitochondrial sequences and one nuclear ribosomal gene have lent support to this hypothesis (Bauzà-Ribot et al. 2012).
  
  
  RESULTS
  We present the analyses of mitochondrial genome sequences of 21 metacrangonyctids in the genera Metacrangonyx and Longipodacrangonyx, covering the entire geographical range of the family. Most mitogenomes were attained by next-generation sequencing techniques using long-PCR fragments sequenced by Roche FLX/454 or GS Junior pyro-sequencing, obtaining a coverage depth per nucleotide of up to 281×. All mitogenomes were AT-rich and included the usual 37 genes of the metazoan mitochondrial genome, but showed a unique derived gene order not matched in any other amphipod mitogenome. We compare and discuss features such as strand bias, phylogenetic informativeness, non-synonymous/synonymous substitution rates and other mitogenomic characteristics, including ribosomal and transfer RNAs annotation and structure.
  
  
  CONCLUSIONS
  Next-generation sequencing of pooled long-PCR amplicons can help to rapidly generate mitogenomic information of a high number of related species to be used in phylogenetic and genomic evolutionary studies. The mitogenomes of the Metacrangonyctidae have the usual characteristics of the metazoan mitogenomes (circular molecules of 15,000-16,000 bp, coding for 13 protein genes, 22 tRNAs and two ribosomal genes) and show a conserved gene order with several rearrangements with respect to the presumed Pancrustacean ground pattern. Strand nucleotide bias appears to be reversed with respect to the condition displayed in the majority of crustacean mitogenomes since metacrangonyctids show a GC-skew at the (+) and (-) strands; this feature has been reported also in the few mitogenomes of Isopoda (Peracarida) known thus far. The features of the rRNAs, tRNAs and sequence motifs of the control region of the Metacrangonyctidae are similar to those of the few crustaceans studied at present.
authors.affiliation: IMEDEA (CSIC-UIB), Mediterranean Institute for Advanced Studies, c/Miquel Marquès 21, 07190 Esporles, Spain. jpons@imedea.uib-csic.es.
authors.forename: Joan
authors.initials: J
authors.lastname: Pons
authors.forename: Maria M
authors.initials: MM
authors.lastname: Bauzà-Ribot
authors.forename: Damià
authors.initials: D
authors.lastname: Jaume
authors.forename: Carlos
authors.initials: C
authors.lastname: Juan
mesh_terms: D033304:Amphipoda / Q000235:genetics*; D000818:Animals; D005075:Biological Evolution; D002874:Chromosome Mapping; D018387:Codon, Initiator; D018388:Codon, Terminator; D054629:Genome, Mitochondrial; D059014:High-Throughput Nucleotide Sequencing; D024101:Mitochondrial Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D018387:Codon, Initiator; D018388:Codon, Terminator; D024101:Mitochondrial Proteins; D012335:RNA, Ribosomal; D012343:RNA, Transfer