First Record of the Brachiopod Lingula Reevei (Brachiopoda, Lingulidae) from the Sea of Japan

Kazuki Kanno, Akihiko Koyama, Yoshihisa Kurita Biogeography 20. 128–130. Sep. 20, 2018

First record of the brachiopod Lingula reevei (Brachiopoda, Lingulidae) from the Sea of Japan

Kazuki Kanno1, Akihiko Koyama1, Yoshihisa Kurita2.

1. Fishery Research Laboratory, Kyushu University, 4-46-24 Tsuyazaki, Fukutsu, Fukuoka 811-3304, Japan. 2. Graduate School of Agricultural Science, Tohoku University, 3-1 Mukai, Konorihama, Onagawa, Miyagi 986-2242, Japan.

Abstract: Four species of Lingula (Brachiopoda, Lingulidae) occur around Japan. Although all four species have undergone drastic decreases in population size, there is little information on their biology, such as their distribution and genetic diversity. Here, we report a Lingula individual collected from the Genkai Sea, in the southern part of the Sea of Japan. The morphological and genetic analysis indicated that this specimen was L. reevei Davidson 1880. This is the first record ofL. reevei collected from the Sea of Japan.

Key words: benthos, distribution, 18S rRNA, Brachiopoda

Introduction vician and has kept its morphology, it is considered a “living fossil” and their evolutionary biology has Brachiopods are members of Spiralia, which in- been studied intensively (Shimizu et al., 2017). The cludes mollusks and annelids. They are characterized species are also important traditional local foods morphologically by calcified bivalves. While bivalve in some regions of Japan. In the nearshore areas of shells are secreted from dorsal to lateral, brachiopod Japan, the populations of all Lingula species have shells are secreted from dorsal to ventral. Lingula is decreased drastically (Kuramochi et al., 2001), and L. a genus of brachiopod, which is distributed in near- anatina and L. adamsi are threatened with extinction shore areas from temperate to trophic zones world- because of habitat degradation (Henmi, 2012). The wide, except in North and South America (Emig, other species L. reevei and L. rostrata are found in 1997). Four Lingula species occur in Japan: Lingula a few localities, thus their biological information is anatina Lamarck, 1801, Lingula reevei Davidson restrictive. Further biological information on Lingula 1880, Lingula adamsi Dall, 1873, and Lingula ros- species is important for understanding their current trata (Shaw, 1798). These inhabit sandy or muddy state, including distributional and genetic data. nearshore and tidal flat sediments (Kuramochi et Here, we report a specimen Lingula sp. collected al., 2001). Because Lingula emerged in the Ordo- from the nearshore of the Genkai Sea, in the southern part of the Sea of Japan. Using morphological ——————————————————————— and genetic data, the specimen was identified as L. *Corresponding author: Yoshihisa Kurita reevei, making this the first record from the Sea of Tel: +81-225-53-2436 Fax: +81-225-53-2303 Japan. E-mail: [email protected]

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Materials and Methods 20s, and 72°C for 30s), with a final 5-min extension at 72°C. Sequencing was conducted by an external Using a dredge, one specimen was collected from service (FASMAC). sandy sediment in the Genkai Sea (water depth approximately 25 m; 33.736°N, 130.409°E), on 3 Au- Result and Discussion gust, 2017. We measured its body, shell, and pedicle lengths. The specimen is preserved at the Onagawa One Lingula individual was collected with other Field Center, Tohoku University. benthic invertebrates, such as Astropecten polyacan- DNA was extracted from a small piece of the ped- thus Müller and Troschel, 1842, Temnopleurus sp., icle using a DNeasy Blood & Tissue Kit (QIAGEN). and Branchiostoma japonicum (Willey, 1897). The We used the primers EukF and EukR (Medlin et al. specimen is approximately 35 mm in body length, 23 1988) to amplify a partial fragment of the nuclear mm in pedicle length, and 12 mm in shell length (Fig. DNA 18S ribosomal RNA (LSU) gene. We also used 2). The pedicle was white and the pedicle length an internal primer (Lingula18SFnst 5'-AGCTCG- was approximately 1.9 times longer than the shell TAGTTGGATCTCAGG-3') for sequencing. Poly- length. This is a morphological trait used for species merase chain reaction (PCR) amplification was car- identification (Kuramochi et al., 2012) and matches ried out in 10 μL reaction mixtures containing 5 μL L. reevei, rather than L. anatina. For further species of EmeraldAmp PCR Master Mix (TaKaRa), 1 μM identification, we decided to partially sequence of each primer, and the DNA template. The reaction the LSU gene. A 1732 bp portion was sequenced mixtures were preheated at 95°C for 3 min, followed (GenBank accession number: LC334155) and the by 30 amplification cycles (95°C for 30s, 50°C for sequence was identical to that of L. reevei (GenBank

Figure 1. Sampling site of this study. Black circle indicated the site.

Figure 2. Lingula reevei collected in this study. Scale bar, 5mm.

− 129 − First record Lingula reevii from the Sea of Japan. accession number: AB747096). On the other hand, of Japanese Tidal Flats: Red Data Book of Sea- comparison of the DNA sequence with other Lingula shore Benthos. 233–234. Tokai University Press. species presented minor substitutions as below: vs. L. Kanagawa (in Japanese with English title) anatine (1729/1732 bp, GenBank accession number Hirose, M., Ohtsuka, S., Kondo, Y., Hirabayashi, T., U08331), vs. L. adamsi (1729/1732 bp, GenBank ac- Tominaga, K. & Shimizu N., 2012. Two species cession number U08329), vs. L. rostrata (1722/1726 of brachiopods collected from central part of Seto bp, GenBank accession number AB855774). LSU Inland Sea, Western Japan. Bulletin of the Hiro- gene is usually highly conservative in this taxon shima University Museum, 4: 43–48. (in Japanese (Kuramochi et al., 2012). Although the high similar- with English abstract) ity is also shown in the comparison of sequences, the Hunter, C. L., Krause, E., Fitzpatrick, J., & Kenne- sequences is different from other Lingula species. dy, J., 2008. Current and historic distribution and From these results, we concluded that our specimen abundance of the inarticulated brachiopod, Lingu- was L. reevei. la reevei Davidson (1880), in Kaneohe Bay, Oahu, In Japan, L. reevei has been found in the Ariake Hawaii, USA. Marine Biology, 155: 205–210. Sea, the Seto-Inland Sea, and Pacific Ocean (Kur- Kuramochi, T., Kimura, K. & Fujimoto, K., 2001. amochi et al., 2001; Yoshigo, 2004). The collected Review of genus Lingula (Brachiopoda: Linguli- individual is the first record of L. reevei from the Sea data) in Japan and adjacent waters. Nanki-Seibut- of Japan. Despite L. reevei now is not registered as a su, 43: 112–116. (in Japanese with English title) threaten species, but several studies have concerned Kuramochi, T., Koi, A., Kashihara, K. & Naganuma, decline in L. reevei populations, not only around Ja- T., 2012. Taxonomic revision of Japanese Lingula pan (Yoshigo, 2004; Hirose et al., 2012) but also in anatina with L. reevei (Brachiopoda : Lingulata). other locations, such as Hawaii (Hunter et al., 2008). Biosphere Science, 51: 27–35. (in Japanese with Little is known about the biology of L. reevei. There- English abstract) fore, the accumulation of data, such as those report- Medlin, L., Elwood, H. J., Stickel, S. & Sogin, M. L., ed here, will become help to understand the current 1988. The characterization of enzymatically am- state of L. reevei, and may facilitate conservation plified eukaryotic 16S-like rRNA-coding regions. activity for this species. Gene, 71: 491–499. Shimizu, K., Luo, Y. J., Satoh, N. & Endo, K., 2017. Reference Possible co-option of engrailed during brachiopod and mollusc shell development. Biology Letters, Emig, C. C., 1997. Ecology of inarticulated brachio- 13: 20170254. pods. In Kaesler, R. L. (Ed) Brachiopoda. Trea- Yoshigoh, H., 2004. Lingula reevei (Brachiopoda) tise on Invertebrate Paleontology. Vol. 1, Part H: collected from the tidal zone of the Takehara City, 473–495. Hiroshima Prefecture, with notes for amounts of Geological Society of America and University of Branchiostoma belcheri. HIBAKAGAKU, 214: Kansas. Kansas. 1–5. (in Japanese with English title) Henmi, Y., 2012 Brachiopods. In Japanese Asso- ciation of Benthology (Ed) Threatened Animal (Received November 22, 2017; Accepted April 5, 2018)

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