Aquatic Insects
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This article was downloaded by:[Umea University Library] On: 23 August 2007 Access Details: [subscription number 781079123] Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK Aquatic Insects International Journal of Freshwater Entomology Publication details, including instructions for authors and subscription information: http://www.informaworld.com/smpp/title~content=t713817864 Morphological and molecular species delimitation within the Holarctic Ilybius angustior complex with a focus on Beringia (Coleoptera: Dytiscidae) Online Publication Date: 01 September 2007 To cite this Article: Nilsson, Anders N. and Ribera, Ignacio (2007) 'Morphological and molecular species delimitation within the Holarctic Ilybius angustior complex with a focus on Beringia (Coleoptera: Dytiscidae)', Aquatic Insects, 29:3, 159 - 171 To link to this article: DOI: 10.1080/01650420701383136 URL: http://dx.doi.org/10.1080/01650420701383136 PLEASE SCROLL DOWN FOR ARTICLE Full terms and conditions of use: http://www.informaworld.com/terms-and-conditions-of-access.pdf This article maybe used for research, teaching and private study purposes. Any substantial or systematic reproduction, re-distribution, re-selling, loan or sub-licensing, systematic supply or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material. © Taylor and Francis 2007 Aquatic Insects September 2007; 29(3): 159 – 171 Morphological and molecular species delimitation within the Holarctic Ilybius angustior complex with a focus on Beringia (Coleoptera: Dytiscidae) ANDERS N. NILSSON1 & IGNACIO RIBERA2 1Department of Ecology and Environmental Science, Umea˚ University, Umea˚, Sweden, and 2Departamento de Biodiversidad y Biologı´a Evolutiva, Museo Nacional de Ciencias Naturales, Madrid, Spain Abstract A fourth species of the Holarctic Ilybius angustior complex, Ilybius minakawai n. sp., is described from the Island of Sakhalin in the Russian far east. Male genitalia are diagnostic within this species complex, Downloaded By: [Umea University Library] At: 08:31 23 August 2007 although from body size and shape the new species cannot be separated from I. churchillensis Wallis from north Alaska and the Canadian low arctic. The small and narrow body characteristic of these two species represents an extreme of the cold climate form of the widespread I. angustior (Gyllenhal). Low variation within the studied large ribosomal and cytochrome c oxidase subunit I mitochondrial genes suggest that speciation events within the I. angustior-complex are fairly recent. In combination with known geographical ranges, the low genetic variation within this species complex suggest speciation within Pleistocene refugia including Beringia. Keywords: Diving beetles, Ilybius angustior, new species, Holarctic, mitochondrial DNA, species delimitation, Sakhalin Introduction The I. subaeneus-group of the genus Ilybius Erichson currently includes 32 species of which 18 are Palearctic, nine Nearctic and five Holarctic (Nilsson 2001). The species occurring in North America were revised by Larson (1987), and the West Palearctic species are more or less well-known (e.g. Nilsson & Holmen 1995). In contrast, many East Palearctic species remain poorly known. Nilsson (1994, 1999) revised the so-called I. crassus- and I. poppiusi- complexes, and we will here describe a new species in the Holarctic I. angustior-complex. The I. angustior-complex is very similar to the closely related I. poppiusi-complex (Ribera et al. 2004, represented by I. angustior (Gyllenhal) and I. anjae Nilsson, respectively), sharing the combination of ridged metatarsomeres and a carinate and rugose sternum 6 in males (Nilsson 1999). The two main differences are found in the smaller body size, and in the partly infuscate distal antennomeres of the species in the I. angustior-complex. Correspondence: Anders N. Nilsson, Department of Ecology and Environmental Science, Umea˚ University, S - 901 87 Umea˚, Sweden. E-mail: [email protected] ISSN 0165-0424 print/ISSN 1744-4152 online ª 2007 Taylor & Francis DOI: 10.1080/01650420701383136 160 A. N. Nilsson & I. Ribera The Holarctic distribution of I. angustior has been recognised already by Sharp (1882). A second species of the complex was described from North America by Kirby (1837) as Colymbetes picipes. Kirby’s species was synonymised with I. angustior by Branden (1885), and not until recently restored as a valid species with a Holarctic distribution (Persson 1985). The third species of the complex, the Nearctic I. churchillensis Wallis, was described from the Canadian low arctic (Wallis 1939). Species limits within this complex have been analysed chiefly by Larson (1987) and Larson et al. (2000) based on morphology. The three known species are poorly differentiated and difficult to separate. We will here describe a fourth species of the I. angustior-complex from the Sakhalin Island in the Russian far east, and provide mitochondrial DNA data for four of the six species of the two complexes. A single male of the new species was collected by S.K. Kholin in 1995, and a larger series was included in the material from the 2002 and 2003 expeditions to Sakhalin recently sent to the senior author by N. Minakawa. We also advance some hypotheses concerning the geographical and temporal origin of the species of the complex, based on the DNA sequence data. Material and methods Morphological analyses Body length was measured from the anterior margin of the head capsule to the apex of the elytra. Width of metasternum (WS) was measured inside marginal bead. Penis length was measured as in Nilsson (1999, Figure 15), and penis width was measured as the maximum Downloaded By: [Umea University Library] At: 08:31 23 August 2007 width of the penis shaft in lateral view. All statistical analysis were run using software Statistica 6.0. Longitude and latitude are given for all mapped localities. (CAN) Coll. Anders N. Nilsson, Umea˚ (CDB) Coll. David Bilton, Plymouth (CLH) Coll. Lars Hendrich, Berlin (FMNH) Finnish Museum of Natural History, Helsinki (IBPV) Institute of Biology and Pedology, Vladivostok (MNCN) National Museum of Natural Science, Madrid (NHRS) Swedish Museum of Natural History, Stockholm (ZIN) Zoological Institute, Academy of Sciences, St. Petersburg The following material of I. angustior was studied morphometrically and genitalia of all males were dissected: Sweden, province Ha¨rjedalen, Flatruet, 2.vii.2001, 5 < 5 ,, leg. J. Bergsten, CAN. - Russia: Siberia, Ob-Jenisej Exp. 1876 F. Trybom, NHRS: Nikandrovski insula, 708400N828500E, 1 < 1 ,; Saostrov, 698450N838400E, 2 < 2 ,; Dudinka 698250N868100E, 1 <; Yakutia, Olene¨k, 688320N 1128170E, 18 – 19.vii.1970, 1 <, leg. R. Angus, CAN; Irkutsk, Tibelti, 518460N 1038140E, 8 – 15.vi.1970, 1 ,, leg. R. Angus, CAN; Siberia, Baikal, Mys Krestovskij, 18 km S Jelantsy, 528390N 1068230E, 25.vi.1993, 2 < 2 ,, leg. Krause & O. Ja¨ger, CAN; Buryatia, Kiakhta district, 5 km NO Kiran village, 508200N 1068530E, 20 – 30.vi.1999, 1 <, leg. I. Melnik, CLH; Yakutia, Kolyma delta, Pokhodskaya Edoma, 698040N 1608570E, 1994, 1 < 1 ,, leg. A. Hobaek, CDB; Yakutia, Srednekolymsk, 678270N 1538400E, 18.vi.1991, 1 <, CLH; Sakhalin, Okha district, Beregovye Langry, 538020N 1418550E, 30.ix.1995, 1 <, leg. S.K. Kholin, CAN; Kamchatka, Petropavlovsk, 538020N 1588380E, 10.vii.1997, 1 <, leg. S.K. Kholin, CAN; Kamchatka, ponds inland from the bay between Cape Zheltyi (south) and Cape Ilya (north), 518330N, 1578430E, 27.vii.1999, 5 < 5 ,, leg. N. Minakawa & K.L. Kurowski, CAN; North Kurils, Paramushir, inland from W base of Vasil’yeva Peninsula, 508010N 1558240E, The Holarctic Ilybius angustior complex (Coleoptera: Dytiscidae) 161 16.viii.1997, 1 < 1 ,, leg. N. Minakawa, CAN; North Kurils, Onekotan, ponds on the plateau inland from Nemo Bay, 498360N, 1548490E, 24.vii.1999, 1 <, leg. N. Minakawa & K.L. Kurowski, CAN. Morphometric data on Nearctic I. angustior (48 < 45 ,) and I. churchillensis (44 < 51 ,) corresponding to Figure 74 in Larson (1987) were received from Dr. D.J. Larson. Molecular methods Genomic DNA was obtained through a standard phenol-chloroform extraction using abdominal tissue of specimens collected in ethanol. Two fragments of mitochondrial genes were sequenced, the large ribosomal unit (rrnL) (ca. 500 bp) and the 30 end of the cytochrome c oxidase subunit I (cox1) (ca. 800 bp) (see Ribera et al. 2003 for details of the primers and conditions of the amplification). An additional fragment was sequenced for some of the specimens (Table I), spanning the 30 end of the rrnL gene, the tRNA-Leu and the 50 end of the gene NADH dehydrogenase subunit 1 (ND1), with the same forward primer as the small rrnL fragment and the reverse primer M223 (Simon et al. 1994) (30-GGTCCCTTACGAA TTTGAATATATCCT-50), amplifying a total of ca. 800 bp. Both forward and reverse sequences were obtained for each fragment, which were assembled and edited using Sequencher 4.2 (GeneCodes Corporation). New sequences