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Diptera, Chironomidae) from the Severnaya Zemlya Archipelago and the Vaigach Island (Russian Arctic

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Diptera, Chironomidae) from the Severnaya Zemlya Archipelago and the Vaigach Island (Russian Arctic Zootaxa 4802 (3): 587–600 ISSN 1175-5326 (print edition) https://www.mapress.com/j/zt/ Article ZOOTAXA Copyright © 2020 Magnolia Press ISSN 1175-5334 (online edition) https://doi.org/10.11646/zootaxa.4802.3.13 http://zoobank.org/urn:lsid:zoobank.org:pub:3CE381FB-959E-4256-8BF0-8178DAC60AEC Morphological description and DNA barcoding of some Diamesinae (Diptera, Chironomidae) from the Severnaya Zemlya Archipelago and the Vaigach Island (Russian Arctic) ANDREY B. KRASHENINNIKOV1,2,3,6, EUGENYI A. MAKARCHENKO4,*, ALEXANDER A. SEMENCHENKO5, MARIA V. GAVRILO3,7 & KRISTINA A. VSHIVKOVA2,8 1Institute of Biological Problems of the North Far East Branch of the Russian Academy of Sciences, Portovaya St. 18, 685000 Magadan, Russia 2Perm State University, Bukireva St., 15 614990 Perm, Russia 3Association “Maritime Heritage: Explore & Sustain”, Icebreaker “Krassin”, The Lieutenant Schmidt emb., 23 Line, 199106 Saint- Petersburg, Russia 4Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far East Branch of the Russian Academy of Sciences, 100 let Vladi- vostoku 159, 690022 Vladivostok, Russia. �[email protected]; http://orcid.org/0000-0003-2765-8729 5Laboratory of Ecology and Evolutionary Biology of Aquatic Organisms, School of Natural Sciences, Far Eastern Federal University, Suhanova St. 8, 690950 Vladivostok, Russia. �[email protected]; https://orcid.org/0000-0001-7207-9529 6 �[email protected]; http://orcid.org/0000-0001-9876-1008 7 �[email protected]; https://orcid.org/0000-0002-3500-9617 8 �[email protected]; https://orcid.org/0000-0002-2472-7429 *Corresponding author Abstract Chironomids of the Diamesinae subfamily from the Russian Arctic were studied using both morphological characters and molecular data. Adult males of Diamesa urvantsevi sp. nov., D. amplexivirilia Hansen, Arctodiamesa appendiculata (Lundström) from Severnaya Zemlya Archipelago and D. arctica (Boheman), Pseudokiefferiella sp. from Vaigach Island were described, redescribed, annotated and figured. A reference 658 bp barcode sequence from a fragment of the mitochondrial gene cytochrome oxidase I (COI) was used as a tool for species delimitation. For D. arctica (Boheman) and Pseudokiefferiella sp. close DNA barcodes from Norway were performed, which allowed to relate these specimens to the described species. Comparisons with corresponding regions of COI between each described species and close related congeneric species produced K2P genetic distances of 0.11–0.16, values well associated with interspecific variation. Phylogenetic relationships for genera Arctodiamesa Makarchenko and Pseudokiefferiella Zavřel were reconstructed for the first time. Key words: Diptera, Chironomidae, Diamesini, taxonomy, DNA barcoding, Russian Arctic Introduction To date, according to preliminary data 32 chironomid species of the subfamily Diamesinae, belonging to 10 gen- era for the territory of the Russian Arctic are known (Kieffer 1922, 1923; Lundström 1915; Zasypkina et al. 1996; Zelentsov & Shilova 1996; Sæther 2004; Zelentsov 2007; Krasheninnikov 2014; Krasheninnikov & Gavrilo 2014; Krasheninnikov & Bogomolova 2017; Makarchenko & Makarchenko 2017). However, the chironomid fauna of most Arctic islands has been studied extremely inadequately. So, only 13 species are known for the Severnaya Zemlya Archipelago of which three species belong to the Diamesinae (Makarchenko 1984; Zelentsov 2007). And for Vaygach Island, only the data of Holmgren (1883) about the discovery of several species of Orthocladiinae are available. This article is devoted to the description, redescription and annotate of the adult males of Diamesa urvantsevi sp. nov., D. amplexivirilia Hansen, Arctodiamesa appendiculata (Lundström) from Severnaya Zemlya Archipelago and D. arctica (Boheman), Pseudokiefferiella sp. from Vaigach Island. Accepted by B. Rossaro: 5 Jun. 2020; published: 24 Jun. 2020 587 In this study the mitochondrial cytochrome c oxidase I (COI) gene 650-bp in size was used as the discriminat- ing molecular marker. This marker has been identified as the core of a global bioidentification system at the species level (Hebert et al. 2003) and has proved to be useful in the Diamesinae (Makarchenko & Semenchenko 2014; Montagna et al. 2016a). For two species, D. arctica (Boheman) and Pseudokieffireella sp., close DNA barcodes from Norway were performed, which allowed to relate these specimens to the described species. Comparisons with corresponding regions of COI between each described species and close related congeneric species produced K2P genetic distances of 0.11–0.16, values well associated with interspecific variation. Phylogenetic relationships for genera Arctodiamesa Makarchenko and Pseudokiefferiella Zavřel were reconstructed for the first time using ob- tained data and available sequences in BOLD systems. Materials and methods The adults of chironomids were preserved in 96% ethanol for DNA-analysis and kept cool at 5°C until further pro- cessing, and in 70% ethanol for study of morphology. The material was slide-mounted in polyvinyl lactophenol and in sandarac medium without chloral hydrate (Krasheninnikov 2011). The morphological terminology and abbrevia- tions used below generally follow Sæther (1980). For some structures of the hypopygium, however, the terminology of Hansen & Cook (1976) and Oliver (1989) and notes on the nomenclature of male genitalia of the genus Diamesa (Montagna et al. 2016b) are used. The photographs were taken using an Axio Lab. A1 (Karl Zeiss) microscope. Holotype of the new species is deposited at the Federal Scientific Center of the East Asia Terrestrial Biodiver- sity, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia (FSCEATB FEB RAS). Paratypes of the new species as well as all other material is stored at the Institute of Biological Problems of the North, Far East Branch of the Russian Academy of Sciences, Magadan, Russia (IBPN FEB RAS, CCK). Genomic DNA was extracted from thorax and abdomen following the protocol of the Qiagen DNeasy Blood & Tissue Kit. The cytochrome oxidase subunit I (CO1) gene was selected for studying. PCR was performed in a 10–25μL reaction volume using Go Taq Green Master Mix (Promega Corp, Madison, WI, USA) or custom mix consist of 1.25 U Platinum ® Taq High Fidelity DNA Polymerase (Invitrogen), 1×PCR buffer, 2.5 mM MgCl2 and 1mM dNTPs. We add 1.25 μM of each primer, 3μL of the respective genomic DNA extract and in some cases added 0.06 μL of BSA (Bovine Serum Albumin solution, concentration 20 mg/mL). PCR and Sanger sequencing used the ‘Folmer primer set’ (LCO1490 + HCO2198) (Folmer et al. 1994). The general PCR profile consisted of an initial denaturation step at 94º C for 2 min, followed by 35 cycles at 94º C for 1 min, 45º C for 30 s, and 72º C for 2 min, and a final extension step of 10 min at 72º C. Amplification of PCR products was checked by electrophoresis using a 1.5% agarose gel in TBE buffer stained with Ethidium bromide and visualized on GelDoc XR+ imaging systems (BioRad). PCR products were purified using ExoSAP-IT (USB Corporation, Cleveland, Ohio, USA) or mix of Exonucle- ase I and Thermosensitive Alkaline Phosphatase (ThermoFisher Scientific, Waltham, MA, USA). Sequencing reac- tion was performed using BigDye® Terminator v3.1 Cycle Sequencing Kits (Applied Biosystems, Carlsbad, CA, USA) and run on an ABI 3130xl Genetic Analyzer Sequencer in Far Eastern Federal University, Vladivostok or on an ABI 3730 in DNA lab of the Natural History Museum, University of Oslo. Phylogenetic analyses were conducted using maximum likelihood (ML) and Bayesian inference (BI) analyses. PartitionFinder 2.1.1 (Lanfear et al. 2012) was used to select the best-fit partitioning scheme and models for BI separately for each codon position of COI. K80 (Kimura 1980), F81 (Felsenstein 1981) and HKY+G (Hasegawa et al. 1985) was the best models of nucleotide substitution for the first, second and third of COI codon position respectively on the reconstructed tree of genus Arctodiamesa. For phylogenetic tree of genus Pseudokiefferiella the best models were SYM+G (Zharkikh 1994), F81 and HKY+G for three COI codon positions. The ML tree was constructed in RAxML v. 8.2.4 using GTR+G model and bootstrapping with 1000 replications (Stamatakis 2006) as well as Bayesian analyses was performed in MrBayes v. 3.2.7 (Ronquist & Huelsenbeck 2003) using 5 000 000 generations, sampled every 100 generations and the first 1000 trees were discarded as burn-in. Moreover, trace files were visually inspected in Tracer 1.7 (Rambaut et al. 2018) and then the consensus tree was visualized in FigTree v. 1.4.4. The neighbor-joining tree of genus Diamesa was reconstructing in MEGA 7 (Kumar et al. 2016). The obtained sequences of described species have been deposited in GenBank under numbers: Diamesa arctica (Boheman)—MK592905 and MK592907, D. urvantsevi sp. nov.—MT367336–MT367337, D. amplexivirilia Han- 588 · Zootaxa 4802 (3) © 2020 Magnolia Press KRASHENINNIKOV ET AL. sen—MT367335, Arctodiamesa appendiculata (Lundström) (MT367334) and Pseudokiefferiella sp.—MT367333. To obtain a representative tree of the genus Arctodiamesa Makarchenko we deposed in GenBank two additional species from Far East—Arctodiamesa marinae Makarchenko (MT452263–MT452267) and A. breviramosa Ma- karchenko (MT452268). Descriptions Diamesa urvantsevi Krasheninnikov et Makarchenko, sp. nov. http://zoobank.org/NomenclaturalActs/2106E8A2-D575-4435-981C-8D4B844676C9 (Figs. 1–4, 8–10) Type material. Holotype, adult male, RUSSIA: Severnaya Zemlya
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