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Drosophila lini  and its two sibling species,  D. ohnishii  and  D. ogumai , are hardly distinguishable from one another in morphology. These species are more or less reproductively isolated. The mitochondrial  ND2  and  COI-COII  and the nuclear ITS1-ITS2 regions were sequenced to seek for the possibility of DNA barcoding and to reconstruct the phylogeny of them. The character-based approach for DNA barcoding detected some diagnostic nucleotides only for monophyletic  D. ogumai , but no informative sites for the other two very closely species,  D. lini  and  D. ohnishii , of which strains intermingled in the molecular phylogenetic trees. Thus, this study provides another case of limited applicability of DNA barcoding in species delineation, as in other cases of related  Drosophila  species. The molecular phylogenetic tree inferred from the concatenated sequences strongly supported the monophyly of the cluster of the three species, that is, the  lini  clade. We propose some hypotheses of evolutionary events in this clade.

DNA Barcoding and Molecular Phylogeny ofDrosophila liniand Its Sibling Species