Survey of the efficacy of a short fragment of the rbcL gene as a supplemental DNA barcode for diatoms |
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Authors: | MacGillivary Michael L Kaczmarska Irena |
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Affiliation: | Department of Biology, Mount Allison University, 63B York Street, Sackville, New Brunswick, E4L 1G7, Canada. mlmcgll@mta.ca |
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Abstract: | DNA barcoding is a tool that uses a short, standard segment of DNA to identify organisms. In diatoms, a consensus on an appropriate DNA barcode has not been reached, but several markers show promise. These include the 5.8S gene plus a fragment of the internal transcribed spacer 2 (ITS‐2) of nuclear‐encoded ribosomal RNA, a 420‐bp segment of the 18S rRNA gene, and a 748‐bp fragment at the 3′‐end of the ribulose bisophosphate carboxylase large subunit (rbcL) gene. Here, we tested a 540‐bp fragment 417‐bp downstream of the start codon of the rbcL gene for its efficacy in distinguishing diatom species in a wide range of taxa. Overall, 381 sequences representing 66 genera and 245 species from the classes Mediophyceae and Bacillariophyceae were examined. Intra/interspecific thresholds were set at p = 0.01 differences per site (diff./site) for Mediophyceae and p = 0.02 diff./site for Bacillariophyceae and correctly segregated 96% and 93% of morphological congeners, respectively. When testing reproductively isolated or biological species, which are only available from Bacillariophyceae, 80% of species were discriminated. Therefore, we concluded that, alone, the rbcL region tested herein as potential a DNA barcode was not a sufficient discriminator of all diatoms. We suggest that this fragment could be used in a dual‐locus barcode with the more variable 5.8S+ITS‐2 to discriminate species without sufficient interspecific divergences in the tested rbcL region and to provide insight into species identity from a separately evolved genome. |
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Keywords: | Biological species divergence threshold interspecific intraspecific morpho‐species p‐distance |
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