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Two New Localities for Piseinotecus Soussi Tamsouri, Carmona, Moukrim & Cervera, 2014 Along the Tyrrhenian Coast

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Two New Localities for Piseinotecus Soussi Tamsouri, Carmona, Moukrim & Cervera, 2014 Along the Tyrrhenian Coast Turkish Journal of Zoology Turk J Zool (2019) 43: 287-289 http://journals.tubitak.gov.tr/zoology/ © TÜBİTAK Short Communication doi:10.3906/zoo-1809-18 Less rare than we thought: two new localities for Piseinotecus soussi Tamsouri, Carmona, Moukrim & Cervera, 2014 along the Tyrrhenian coast Giulia FURFARO*, Paolo MARIOTTINI Department of Science, University of Roma Tre, Rome, Italy Received: 14.09.2018 Accepted/Published Online: 13.03.2019 Final Version: 06.05.2019 Abstract: The geographic range of Piseinotecus soussi Tamsouri, Carmona, Moukrim & Cervera, 2014 is here updated, since it was found in two new localities along the Tyrrhenian coast of the Mediterranean Sea. Furthermore, to confirm the identity of specimens recorded, a DNA barcoding approach was carried out using mitochondrial cytochrome c oxidase subunit I (COI) as it is the molecular marker mostly used in barcoding analysis in nudibranchs. Key words: Piseinotecidae, Tyrrhenian Sea, cryptic diversity, rare species, DNA barcoding Cryptic diversity has caused an underestimation preserved in 95% EtOH for further molecular analysis, of marine biodiversity in many groups, including the and stored in the Department of Science of Roma Tre nudibranch fauna of the Mediterranean Sea. Recently, University. These rare sea slugs were found crawling on molecular approaches have helped to unravel cryptic hard substrata in a coralligenous habitat: two specimens species, revealing that morphological traits can often be at 15 m depth, at Framura, Liguria (Northern Tyrrhenian severely misleading in nudibranchs (Furfaro et al., 2016a, Sea: 41°11′58.3″N, 9°33′20.6″E), and one at 27 m depth, 2016b, 2016c). The aeolid genus Piseinotecus Er. Marcus, at Torre Fico, Cape Circeo (Central Tyrrhenian Sea: 1955 belongs to the family Piseinotecidae Edmunds, 41°13′05.6″N, 13°05′26.5″E). 1970 and includes four species worldwide, of which two The species identity of the Italian specimens was are represented in the Mediterranean: P. soussi Tamsouri, investigated by a DNA barcoding approach using the Carmona, Moukrim & Cervera, 2014, described from the COI mitochondrial marker. DNA was extracted from Atlantic coast of Morocco and the southwestern Iberian four specimens (Table) using the salting-out procedure Peninsula and recorded in Italy only from the Ionian (Aljanabi and Martinez, 1997). Partial COI sequences Sea (Apulia) (Tamsouri et al., 2014) (Figure 1), and P. were amplified by polymerase chain reaction (PCR), sphaeriferus (Schmekel, 1965), originally described from setting the parameters according to Furfaro et al. (2016c), the Gulf of Naples and recorded only three times in the using the universal primers LCO1490 and HCO2198 whole Mediterranean Sea. Piseinotecus soussi is a small (Folmer et al., 1994). Amplicons were sequenced by the aeolid phenotypically indistinguishable from Edmundsella European Division of Macrogen Inc. (Amsterdam, the albomaculata (Pola, Carmona, Calado & Cervera, 2014), Netherlands). A BLASTN (Altschul et al., 1990) search was an extra-Mediterranean flabellinid known only from São conducted to rule out sample contamination. Specimens Vicente Island (Cape Verde), and also very similar to collected are stored at the Department of Science of Edmundsella pedata (Montagu, 1816), a very common Roma Tre University. New sequences are deposited in the flabellinid Mediterranean species from which it differs European Nucleotide Archive (ENA) (http://www.ebi. by the presence of a variable number of white dots on the ac.uk/ena/data/view/PRJEB31616). Sequences obtained cerata. In the framework of an intensive field survey on were aligned with sequences of P. soussi from the type the nudibranch fauna of the Tyrrhenian coasts, we have locality, retrieved from GenBank (Table). The maximum found three specimens of P. soussi (vouchers: RM3_1356, p-distance, calculated using MEGA 6.0 (Tamura et al., RM3_1357, RM3_1408) (Figure 2) from two additional 2013), was 0.5%, confirming all the Italian individuals localities (red squares in Figure 1). These individuals so far collected as belonging to P. soussi (Figure 2). This were photographed both in situ and in the laboratory, is the first study focusing on this Piseinotecus species * Correspondence: [email protected] 287 This work is licensed under a Creative Commons Attribution 4.0 International License. FURFARO and MARIOTTINI / Turk J Zool Figure 1. Map showing the current known range of Piseinotecus soussi: previous records (blue circles) and new ones (red squares). 1: Original description of the species by Tamsouri et al. (2014); 2: OPK at https://opistobranquis.info/ca/guia/nudibranchia/cladobranchia/ aeolidioidea/piseinotecus-soussi/; 3: Framura, 41°11′58.3″N, 9°33′20.6″E, present study; 4: Circeo Promontory, 41°13′05.6″N, 13°05′26.5″E, present study. Figure 2. Italian specimens of P. soussi. A, B) Specimens from Ligurian Sea: (A) RM3_1357 and (B) RM3_1356. C, D) Specimen (RM3_1408) from Cape Circeo Latium: (C) in situ and (D) in aquarium. Scale bar = 2 mm. 288 FURFARO and MARIOTTINI / Turk J Zool Table. Species names, collection localities, voucher IDs, GenBank accession numbers, and references of the P. soussi specimens analyzed in this work. Species Locality Voucher Accession number Reference Piseinotecus soussi Santa Maria, Apulia (Ionian Sea) RM3_862 LR535941 Present study Piseinotecus soussi Santa Maria, Apulia (Ionian Sea) RM3_863 LR535942 Present study Piseinotecus soussi Framura, Liguria (Tyrrhenian Sea) RM3_1356 LR535943 Present study Piseinotecus soussi Circeo Promontory, Latium (Tyrrhenian Sea) RM3_1408 LR535944 Present study Piseinotecus soussi W Andalusia, Spain (Atlantic Ocean) BAU2952 LT718575 Furfaro et al., 2018 Piseinotecus soussi W Andalusia, Spain (Atlantic Ocean) BAU2953 LT718576 Furfaro et al., 2018 by molecular data, and it also confirms that the assayed benthic fauna, including focused field surveys and an specimens from Apulia, which were reported as P. soussi integrative taxonomic approach, is mandatory to fill the in the original description of this species without any gaps caused by the cryptic diversity in the Mediterranean anatomical investigation, actually belong to it and not Sea. to the phenotypically indistinguishable E. albomaculata or the similar E. pedata. This is not trivial, considering Acknowledgments the steadily growing number of cryptic species with The authors sincerely acknowledge the University of Roma Atlantic versus Mediterranean ranges revealed by genetic Tre for financial support (CAL/2017). The authors wish to investigations. thank Fabio Vitale (Lecce, Apulia) for providing samples The new records of this rare species suggest that the from the type locality and the two anonymous referees geographical distribution of this species might be wider who helped improve the manuscript. GF wish to thank than previously reported, considering its minute size Walter Cutrupi, Matteo Garzia, and Emilio Prosperi and causing its hard spotting in the field, and easy confusion all the staff of La Tribù Diving and Style Diving for their with the strikingly similar and common Mediterranean professional and kind help during the field work. species E. pedata. Special attention to the inconspicuous References Aljanabi SM, Martinez I (1997). Universal and rapid salt-extraction Furfaro G, Picton B, Martynov A, Mariottini P (2016). Diaphorodoris of high quality genomic DNA for PCR-based techniques. alba Portmann & Sandmeier, 1960 is a valid species: molecular Nucleic Acids Res 25: 4692-4693. and morphological comparison with D. luteocincta (M. Sars, 1870) (Gastropoda: Nudibranchia). Zootaxa 4193: 304-316. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990). Basic local alignment search tool. J Mol Biol 215: 403-410. Furfaro G, Salvi D, Mancini E, Mariottini P (2018). A multilocus view on Mediterranean aeolid nudibranchs (Mollusca): systematics Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994). DNA and cryptic diversity of Flabellinidae and Piseinotecidae. Mol primers for amplification of mitochondrial cytochrome c Phylogenet Evol 118: 13-22. oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3: 294-299. Tamsouri N, Carmona L, Moukrim A, Cervera JL (2014). Addenda to the article Bull Mar Sci. 90: 991-997 (2014): description of Furfaro G, Mariottini P, Modica MV, Trainito E, Doneddu M, Oliverio a new species of Piseinotecus (Gastropoda, Heterobranchia, M (2016). Sympatric sibling species: the case of Caloria elegans Piseinotecidae) from the northeastern Atlantic Ocean. Bull and Facelina quatrefagesi (Gastropoda: Nudibranchia). Sci Mar Mar Sci 91: 83-84. 80: 511-520. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013). Furfaro G, Modica MV, Oliverio M, Mariottini P (2016). A MEGA6: Molecular Evolutionary Genetics Analysis version DNA-barcoding approach to the phenotypic diversity of 6.0. Mol Biol Evol 30: 2725-2729. Mediterranean species of Felimare Ev. Marcus & Er. Marcus, 1967 (Mollusca: Gastropoda), with a preliminary phylogenetic analysis. Ital J Zool 83: 195-207. 289.
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