Abstract

Short DNA sequences from a standardized region of the genome provide a DNA barcode for identifying species. Compiling a public library of DNA barcodes linked to named specimens could provide a new master key for identifying species, one whose power will rise with increased taxon coverage and with faster, cheaper sequencing. Recent work suggests that sequence diversity in a 648-bp region of the mitochondrial gene, cytochrome c oxidase I (COI), might serve as a DNA barcode for the identification of animal species. This study tested the effectiveness of a COI barcode in discriminating bird species, one of the largest and best-studied vertebrate groups. We determined COI barcodes for 260 species of North American birds and found that distinguishing species was generally straightforward. All species had a different COI barcode(s), and the differences between closely related species were, on average, 18 times higher than the differences within species. Our results identified four probable new species of North American birds, suggesting that a global survey will lead to the recognition of many additional bird species. The finding of large COI sequence differences between, as compared to small differences within, species confirms the effectiveness of COI barcodes for the identification of bird species. This result plus those from other groups of animals imply that a standard screening threshold of sequence difference (10× average intraspecific difference) could speed the discovery of new animal species. The growing evidence for the effectiveness of DNA barcodes as a basis for species identification supports an international exercise that has recently begun to assemble a comprehensive library of COI sequences linked to named specimens.

Highlights

  • The use of nucleotide sequence differences in a single gene to investigate evolutionary relationships was first widely applied by Carl Woese (Woese and Fox 1977)

  • All 260 bird species had a different c oxidase I (COI) sequence(s); none was shared between species

  • With a few interesting exceptions discussed below, COI sequence differences between closely related species were far higher than differences within species (18fold higher; average Kimura-2-parameter [Kimura-2 parameter (K2P)] differences between and within species, 7.93% and 0.43%, respectively) (Figure 1)

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Summary

Introduction

The use of nucleotide sequence differences in a single gene to investigate evolutionary relationships was first widely applied by Carl Woese (Woese and Fox 1977). He recognized that sequence differences in a conserved gene, ribosomal RNA, could be used to infer phylogenetic relationships. Like rRNA, often do not differ among closely related organisms, but they are indispensable in recovering ancient relationships, providing insights as far back as the origin of cellular life (Woese 2000). Genes that evolve rapidly may overwrite the traces of ancient affinities, but regularly reveal divergences between closely related species

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