The design and testing of mini-barcode markers in marine lobsters

PLoS One. 2019 Jan 24;14(1):e0210492. doi: 10.1371/journal.pone.0210492. eCollection 2019.

Abstract

Full-length mitochondrial cytochrome c oxidase I (COI) sequence information from lobster phyllosoma larvae can be difficult to obtain when DNA is degraded or fragmented. Primers that amplify smaller fragments are also more useful in metabarcoding studies. In this study, we developed and tested a method to design a taxon-specific mini-barcode primer set for marine lobsters. The shortest, most informative portion of the COI gene region was identified in silico, and a DNA barcode gap analysis was performed to assess its reliability as species diagnostic marker. Primers were designed, and cross-species amplification success was tested on DNA extracted from a taxonomic range of spiny-, clawed-, slipper- and blind lobsters. The mini-barcode primers successfully amplified both adult and phyllosoma COI fragments, and were able to successfully delimit all species analyzed. Previously published universal primer sets were also tested and sometimes failed to amplify COI from phyllosoma samples. The newly designed taxon-specific mini-barcode primers will increase the success rate of species identification in bulk environmental samples and add to the growing DNA metabarcoding toolkit.

MeSH terms

  • Animals
  • Biomarkers / analysis*
  • DNA Barcoding, Taxonomic / methods*
  • DNA Primers / genetics
  • Electron Transport Complex IV / genetics*
  • Mitochondrial Proteins / genetics*
  • Nephropidae / classification
  • Nephropidae / genetics*
  • Protein Subunits
  • Reproducibility of Results
  • Seafood
  • Species Specificity

Substances

  • Biomarkers
  • DNA Primers
  • Mitochondrial Proteins
  • Protein Subunits
  • Electron Transport Complex IV

Grants and funding

The authors received no specific funding for this work.