Molluscan Research 29(1): 33–59 ISSN 1323-5818 http://www.mapress.com/mr/ Magnolia Press The phylogeny and taxonomy of New Zealand Notoacmea and Patelloida species (Mollusca: Patellogastropoda: Lottiidae) inferred from DNA sequences TOMOYUKI NAKANO1, BRUCE A. MARSHALL2, MARTYN KENNEDY3 & HAMISH G. SPENCER3 1Department of Geology and Palaeontology, National Museum of Nature and Science, 3-23-1 Hyakunin-cho, Shinjuku-ku, Tokyo 169-0073, Japan 2Museum of New Zealand Te Papa Tongarewa, P.O. Box 467, Wellington, New Zealand 3Allan Wilson Centre for Molecular Ecology and Evolution, Department of Zoology, University of Otago, P.O. Box 56, Dune- din 9054, New Zealand Corresponding author: Hamish G. Spencer ([email protected]) Abstract The systematics of Notoacmea Iredale, 1915 have been confused because of their highly variable shells. We used DNA sequences from a mitochondrial gene (COI) and a nuclear gene (ITS1) to define the species boundaries among New Zealand Notoacmea species, using the allied genus Patelloida Quoy & Gaimard, 1834 as outgroup. Phylogenetic trees of 195 individu- als showed 14 well-supported, reciprocally monophyletic clades, which we treat as species: N. badia Oliver, 1926, N. cella- noides Oliver, 1926, N. daedala (Suter, 1907), N. elongata (Quoy & Gaimard, 1834), N. parviconoidea (Suter, 1907), N. pileopsis (Quoy & Gaimard, 1834), N. scapha (Suter, 1907), N. scopulina Oliver, 1926, N. sturnus (Hombron & Jacquinot, 1841), N. subantarctica Oliver, 1926, Patelloida cortica (Hutton, 1880) and three new Notoacmea species, N. potae, N. rapida, and N. turbatrix, which are described here. Of the above names, N. daedala and N. subantarctica are resurrected from synon- ymy. Notoacmea helmsi (E.A. Smith, 1894) and N. virescens Oliver, 1926 are interpreted as synonyms of N. elongata. Key words: Gastropoda, limpet, new taxa, species boundaries, COI, ITS1, intertidal. Introduction limpet systematics, both for estimating phylogenetic trees (e.g., Koufopanou et al. 1999; Nakano and Ozawa 2004; Limpets are a diverse and ecologically important group of Kirkendale and Meyer 2004; Goldstien et al. 2006), as well molluscs. They are found throughout the world’s oceans, as for clarifying species complexes that are difficult to from tropical to polar regions, and from the deep-sea to well distinguish on morphological characters alone (Simison and above the high tide line. The lottiid limpet genus Notoacmea Lindberg 1999, 2003; Kirkendale and Meyer 2004; Nakano Iredale, 1915 has long been considered to exhibit an and Ozawa 2005). In the case of Notoacmea, Nakano and antitropical distribution, with representative species from Ozawa (2007) showed the monophyly of the genus using temperate waters of both the southern and northern Pacific three mitochondrial genes and Nakano and Spencer (2007) (Kira 1961; Habe and Kosuge 1967; Keen 1971; Abbott utilized both mitochondrial and nuclear genes to discover 1974; Habe and Okutani 1975; Ponder and Creese 1980; that there are five distinct species within the taxon N. helmsi Powell 1979). More recently, however, Lindberg (1986) as currently interpreted. The goal of the present study is to transferred all the North American species to Tectura Gray, elucidate the species boundaries among the New Zealand 1847, and Sasaki and Okutani (1993) erected a new genus, species of Notoacmea using sequences from the Nipponacmea Sasaki & Okutani, 1993, for the Japanese mitochondrial cytochrome c oxidase subunit I gene (COI) species. Nakano and Ozawa (2004) subsequently confirmed and the nuclear internal transcribed spacer 1 (ITS1), and to the monophyly of Nipponacmea, but restricted Tectura to rectify the nomenclature to reflect this phylogeny. The two one northeastern Atlantic species, T. virginea (Müller, 1776), genera dealt with in this paper are readily distinguished on and removed the American Pacific species to Lottia G.B. radular characters; Notoacmea by lack of marginal teeth Sowerby I, 1834. As the result of these revisions, Notoacmea (radular formula 0-2-0-2-0), Patelloida Quoy & Gaimard, now consists solely of species from New Zealand and 1834 by presence of two marginal teeth on each side of each Australian waters. In New Zealand eight Notoacmea species cross row (2-2-0-2-2). and two subspecies are currently recognized (Powell 1979; Spencer et al. 2006); additionally two species and two subspecies have been treated as synonyms. Materials and Methods Historically, taxonomic studies of these limpets have been based on shell morphology and radular characters (e.g., Field observation and collection of samples Suter 1907; Oliver 1926; Powell 1973; Ponder and Creese Table 1 lists the species and collection data (localities 1980), but the highly variable shell morphology of limpets and habitats) of specimens used in this study, representing has led to taxonomic confusion and the failure to recognize eight of the nine currently recognized New Zealand species species complexes (Sasaki 1999; Nakano and Spencer 2007). and subspecies of Notoacmea, as well as one distinctive Molecular techniques have recently proved very useful in named form. In addition, a species of Patelloida was also COPYRIGHT © 2009 MALACOLOGICAL SOCIETY OF AUSTRALASIA & SOCIETY FOR THE STUDY OF MOLLUSCAN DIVERSITY 33 34 NAKANO ET AL. (2009) MOLLUSCAN RESEARCH, VOL. 29 collected to use as an outgroup. Sampling was carried out at 4 ˚C. In total, 145 individuals were newly sequenced, and 39 intertidal shore sites in mainland New Zealand between combined with published sequences of 50 individuals from the Bay of Islands in the North Island and Southland in the Nakano and Spencer (2007). All voucher specimens are South Island, as well as three additional sites from the deposited (database linked to GenBank numbers) in the subantarctic Auckland and Campbell Islands, with type Museum of New Zealand Te Papa Tongarewa (NMNZ). localities of different species included if possible. All Table 1 also shows both the revised species name (the result localities of individuals genetically analyzed are plotted on of this study) along with the NMNZ lot numbers and maps (Figs 1, 2). Living specimens were preserved in 70% GenBank accession numbers for all specimens genetically ethanol and returned the laboratory where they were stored at analyzed in this study. TABLE 1. Species identification, localities, detailed habitats, NMNZ lot numbers and GenBank accession numbers for specimens genetically analyzed in this study. Type localities are indicated with *. NMNZ GenBank Accession No. Species Locality HabitatLot No. COI ITS1 N. badia St. Clair, Dunedin* middle shore, medium exposure tide pool 184115-A AB287009 AB287128 St. Clair, Dunedin* middle shore, medium exposure tide pool 184115-B AB287010 AB287129 Oamaru under rocks on lower shore, medium exposure 184116 AB287011 AB287130 Oamaru under rocks on lower shore, medium exposure 184117 AB287012 AB287131 Katiki under rocks on lower shore, medium exposure 184118 AB287013 AB287132 N. cellanoides South of Cape Campbell on rocks on upper shore 184119 AB287014 AB287133 Kaikoura on rocks on upper shore 184120 AB287015 AB287134 Island Bay, Wellington on rocks on upper shore 184121 AB287016 AB287135 Castlepoint on rocks on upper shore 184122 AB287017 AB287136 Ocean Beach, Whangarei on rocks on upper shore 184123 AB287018 AB287137 N. daedala Vauxhall, Otago Harbour under rocks on lower sheltered shore 184284 AB287037 AB287156 Kaikoura under rocks on lower shore, medium exposure 184290 AB287038 AB287157 Titahi Bay under rocks on lower shore, medium exposure 184289 AB287039 AB287158 Mahia under rocks on lower shore, medium exposure 184286 AB287040 AB287159 Tatapouri, Gisborne under rocks on lower shore, medium exposure 184287 AB287041 AB287160 McLeod Bay, Whangarei Harbour under rocks on middle sheltered shore 184288 AB287042 AB287161 Halls Beach, Waitemata Harbour* under rocks on middle sheltered shore 184285 AB287043 AB287162 N. elongata Harington Point, Otago Harbour under rocks on lower shore, medium exposure 184124-A AB287019 AB287138 Harington Point, Otago Harbour under rocks on lower shore, medium exposure 184124-B AB287020 AB287139 Vauxhall, Otago Harbour on rocks on lower sheltered shore 184125 AB287021 AB287140 Bluff under rocks on lower sheltered shore 184126 AB287022 AB287141 Jackson Bay under rocks on lower shore, medium exposure 184127 AB287023 AB287142 Nelson Haven under rocks on lower sheltered shore 184128 AB287024 AB287143 French Pass* under rocks on lower shore, medium exposure 184129 AB287025 AB287144 Lyttelton Harbour, Christchurch under rocks on lower shore, medium exposure 184130 AB287026 AB287145 Port Levy on rocks on lower sheltered shore 184131 AB287027 AB287146 Pigeon Bay under rocks on lower sheltered shore 184132 AB287028 AB287147 Island Bay, Wellington under rocks, exposed shore 184133 AB287029 AB287148 Castlepoint under rocks, exposed shore 184134 AB287030 AB287149 Maketu under rocks, medium exposure 184135 AB287031 AB287150 North of Cape Egmont under rocks on exposed shore 184136 AB287032 AB287151 Cornwallis, Manukau Harbour under rocks, sheltered shore 184137 AB287033 AB287152 Tapeka Point, Bay of Islands under rocks, medium exposure 184138 AB287034 AB287153 Opononi, Hokianga Harbour under rocks, sheltered shore 184139 AB287035 AB287154 Halls Beach, Waitemata Harbour under rocks on sheltered middle shore 184140-A AB287036 AB287155 ...... continued on the next page PHYLOGENY OF NEW ZEALAND LOTTIIDAE 35 TABLE 1 (contnued) NMNZ GenBank Accession No. Species Locality HabitatLot No. COI