Next-Generation Mitogenomics: A Comparison of Approaches Applied to Caecilian Amphibian Phylogeny

PLoS One. 2016 Jun 9;11(6):e0156757. doi: 10.1371/journal.pone.0156757. eCollection 2016.

Abstract

Mitochondrial genome (mitogenome) sequences are being generated with increasing speed due to the advances of next-generation sequencing (NGS) technology and associated analytical tools. However, detailed comparisons to explore the utility of alternative NGS approaches applied to the same taxa have not been undertaken. We compared a 'traditional' Sanger sequencing method with two NGS approaches (shotgun sequencing and non-indexed, multiplex amplicon sequencing) on four different sequencing platforms (Illumina's HiSeq and MiSeq, Roche's 454 GS FLX, and Life Technologies' Ion Torrent) to produce seven (near-) complete mitogenomes from six species that form a small radiation of caecilian amphibians from the Seychelles. The fastest, most accurate method of obtaining mitogenome sequences that we tested was direct sequencing of genomic DNA (shotgun sequencing) using the MiSeq platform. Bayesian inference and maximum likelihood analyses using seven different partitioning strategies were unable to resolve compellingly all phylogenetic relationships among the Seychelles caecilian species, indicating the need for additional data in this case.

Publication types

  • Comparative Study

MeSH terms

  • Amphibians / classification
  • Amphibians / genetics*
  • Animals
  • Bayes Theorem
  • DNA, Mitochondrial / genetics*
  • Genetic Markers
  • Genetic Variation
  • Genome, Mitochondrial / genetics*
  • High-Throughput Nucleotide Sequencing / methods*
  • Molecular Sequence Annotation
  • Phylogeny
  • Selection, Genetic
  • Sequence Analysis, DNA
  • Seychelles
  • Species Specificity

Substances

  • DNA, Mitochondrial
  • Genetic Markers

Grants and funding

This work was funded by Biotechnology and Biological Sciences Research Council/Natural Environment Research Council (http://www.bbsrc.ac.uk/ and http://www.nerc.ac.uk/), SynTax grant (MW, JJD); Natural History Museum/University College London (NHM/UCL) Impact Scholarship (http://www.nhm.ac.uk/ and http://www.ucl.ac.uk/), PhD studentship (JJD, DJG); Darwin Initiative grant (19-002) (https://www.gov.uk/government/groups/the-darwin-initiative) (DJG, MW, JJD); and Ministry of Economy and Competitiveness of Spain (grants ERDF co-funded CGL2012-40082 and RYC-2011-09321) (http://www.mineco.gob.es/portal/site/mineco?lang_choosen=en-DSM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.