A ragworm specimen was obtained subtidally in Oshoro Bay, Hokkaido, Japan, about 43°12′N, 140°51′E, on 2 June 2014 by Osamu Horiguchi, identified by Hiroshi Kajihara as Nereiphylla hera Kato and Mawatari, 1999, and photographed and fixed in 99% EtOH by Takumi Onishi. Total DNA was extracted from the posterior end of the body using the silica method (Boom et al. 1990) with some modifications. Extracted DNA was dissolved in 30 µl of deionized water and has been preserved at –20°C. Remaining morphological voucher specimen has been deposited at the Hokkaido University Museum under the catalogue number ICHU2120251 (contact: Dr. Hiroshi Kajihara, kazi@mail.sci.hokudai.ac.jp).
PCR amplification was attempted for three gene markers, using the primer pairs LCO1490 (5′-GGTCAACAAATCATAAAGATATTGG-3′) and HCO2198 (5′-TAAACTTCAGGGTGACCAAAAAATCA-3′) (Folmer et al. 1994) for the mitochondrial cytochrome c oxidase subunit I gene, 16S ar-L (5′-CGCCTGTTTATCAAAAACAT-3′), and LSU5 (5′-ACCCGCTGAAYTTAAGCA-3′) and LSU3 (5′-TCCTGAGGGAAACTTCGG-3′) (Littlewood 1994) for the nuclear 28S rRNA gene. PCR products were visualized by electrophoresis in 1% agarose gel, but none of the three gene markers attempted were successfully amplified, probably due to a failure in DNA extraction.
Taxonomy
Phylum Annelida
Family Phyllodocidae Stephen and Edmonds, 1972
Genus Nereiphylla Blainville, 1828
Nereiphylla hera Kato and Mawatari, 1999
(Figs 1, 2)
Fig. 1. Nereiphylla hera Kato and Mawatari, 1999 (ICHU2120251) from Oshoro Bay, Hokkaido, Japan, general shape of body.
Fig. 2. Nereiphylla hera Kato and Mawatari, 1999 (ICHU2120251), magnification of head, dorsal view.
References
Boom, R., Sol, C. J. A., Salimans, M. M. M., Jansen, C. L., Wertheim-van Dillen, P. M. E., and van der Noordaa, J. 1990. Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28: 495–503.
Folmer, O., Black, M., Hoeh, W., Lutz, R. and Vrijenhoek, R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3: 294–299.
Littlewood, D. T. 1994. Molecular phylogenetics ofcupped oysters based on partial 28S rRNA gene sequences. Molecular Phylogenetics and Evolution 3: 221–229.
Palumbi, S., Martin, A., Romano, S., McMillan, W. O., Stice, L., Grabowski, G. 1991. The Simple Fools Guide to PCR, Ver. 2. Department of Zoology and Kewalo Marine Laboratory, University of Hawaii, Honolulu, 45 pp.