Abstract
Morphological analysis of all developmental stages (except female), mitochondrial DNA sequences from cytochrome c oxidase subunit I (cox1) and cytological analysis of the polytene chromosomes were used to describe a new species of Chironomus found in the littoral and profundal zones of an endorheic, warm-monomictic lake in Mexico. Male imago is distinguished by the shape of superior volsella and by an antennal and bristle ratio lower than two. The pupa is characterized by the spur morphology of abdominal segment VIII. There is also a continuous row of hooklets on abdominal segment II. The larva is distinguished by a combination of antenna, mentum, mandible, and pecten epipharyngis characteristics, and abdominal ventral tubules. Molecular and cytological analysis supported the morphological differences found. The maximum likelihood tree obtained shows that Chironomus alchichica sp. n. clusters together with Chironomus decorus-group sp. 2 Butler et al. (1995) (bootstrap support = 92%), but genetic p-distances within C. alchichica sp. n. (0.004) were lower than the p-distances between other species of the decorus-group (C. decorus-group sp. 2, Chironomus bifurcatus Wülker et al., 2009 and Chironomus maturus Johannsen, 1908) confirming that it is a different species. The new species belongs to thummi cytocomplex, (decorus-group), with chromosome set- 2n = 8 and chromosome arm combinations: AB CD EF G. Karyologically, the species is closest to Chironomus riihimaekiensis Wülker (1973). This species has very compact salivary gland chromosomes with well heterochromatinized centromere regions in chromosomes AB CD G. Several fixed homozygous inversions distinguish arm A of the species from that of C. riihimaekiensis. Arm E differs from that of C. riihimaekiensis by simple fixed homozygous inversion. Some similarities in band sequences of this arm were found with species from the decorus-group as Chironomus blaylocki Wülker et al., 2009 and C. bifurcatus (decorus-group). The position of the key constrictions in chromosome G: Nucleolar organizer (NOR) and Balbiani rings (BRs) is similar to the species of decorus-group. C. alchichica sp. n. has been found in soft sediments rich in organic matter in well mineralized waters (where conductivity >10 mS cm-1) and with a high pH (≥9). The profundal zone is inhabited only during the mixing period, when dissolved oxygen is present.
References
Adame, M.F., Alcocer, J. & Escobar, E. (2008) Size-fractionated phytoplankton biomass and its implications for the dynamics of an oligotrophic tropical lake. Freshwater Biology, 53 (1), 22–31.
https://doi.org/10.1111/j.1365-2427.2007.01864.x
Alcocer, J., Lugo, A., Escobar, E., Sánchez, M.R. & Vilaclara, G. (2000) Water column stratification and its implications in the tropical warm monomictic Lake Alchichica, Puebla, México. Verhandlungen des Internationalen Verein Limnologie, 27, 3166–3169.
Alcocer, J. & Bernal-Brooks, F.W. (2010) Limnology in Mexico. Hydrobiologia, 644 (1), 1–54.
https://doi.org/10.1007/s10750-010-0211-1Alcocer, J., Escobar, E., Řezníčková, P. & Oseguera, L.A. (2016) La comunidad de macroinvertebrados bentónicos litorales como un reflejo de la heterogeneidad ambiental. Hidrobiologica, 26 (3), 403–418.
Andersen, F.S. (1949) On the subgenus Chironomus. Studies on the systematics and biology of Chironomidae III. Videnskabelige Meddelser fra Dansk Naturhistorisk Forening, III, 1–66.
Armitage, P.D., Pinder, L. & Cranston, P. (1995) The Chironomidae: Biology and Ecology of Non-biting Midges. Springer, London, 572 pp.
https://doi.org/10.1007/978-94-011-0715-0Beck, E. (1961) Two new Chironomidae (Diptera) and additional state records from Florida. Florida Entomologist, 44 (3), 125–128.
https://doi.org/10.2307/3492971Beneberu, G. & Mengistou, S. (2015) Head capsule deformities in Chironomus spp. (Diptera: Chironomidae) as indicator of environmental stress in Sebeta River, Ethiopia. African Journal of Ecology, 53 (3), 268 –277.
https://doi.org/10.1111/aje.12175Brandon, R.A., Maruska, E.J. & Rumph, W.T. (1981) A new species of neotenic Ambystoma (Amphibia, Caudata) endemic to Laguna Alchichica, Puebla, Mexico. Bulletin of the Southern California Academy of Sciences, 80 (3), 112–125.
Brooks, S.J., Langdon, P.G. & Heiri, O. (2008) The identification and use of Palearctic Chironomidae larvae in palaeoecology. QRA Technical Guide No. 10. Journal of Paleolimnology, 40 (2), 751–753.
https://doi.org/10.1007/s10933-007-9191-1
Butler, M.G., Kiknadze, I.I., Cooper, J.K. & Siirin, M.T. (1995) Cytologically identified Chironomus species from lakes in North Dakota and Minnesota, USA. In: Cranston, P.S. (Ed.), Chironomids, from Gene to Ecosystems. Proceedings of the 12th International Symposium on Chironomidae, Canberra, 23–26 January, 1994. CSIRO, Canberra, pp. 31–37.
Carew, M.E., Pettigrove, V. & Hoffmann, A.A. (2003) Identifying chironomids (Diptera: Chironomidae) for biological monitoring with PCR-RFLP. Bulletin of Entomological Research, 93 (6), 483–490.
https://doi.org/10.1079/ber2003268
Cohuo, S., Hernández, Ma., Pérez, C.L. & Alcocer, J. (2017) Candona alchichica (Podocopida: Candonidae), a new ostracod species from saline, tropical Lake Alchichica, Mexico. Journal of Limnology, 76 (1), 68–84.
https://doi.org/10.4081/jlimnol.2016.1480
Cranston, P.S. (1995) Introduction. In: Armitage, P.D., Cranston, P. & Pinder, L.C. (Eds.), The Chironomidae: Biology and Ecology of Non-biting Midges. Chapman and Hall, London, pp. 1–5.
https://doi.org/10.1007/978-94-011-0715-0_1De Buen, F. (1945) Investigaciones sobre ictiología mexicana. I. Atherinidae de aguas continentales de México. Anales del Instituto de Biología de la Universidad Nacional Autónoma de México, XVI, 475–532.
Drummond, A.J., Ashton, B., Cheung, M., Heled, J., Kearse, M., Moir, R., Stones- Havas, S., Thierer, T. & Wilson, A. (2009) Geneious v.4.6.5. created by Biomatters. [software]
Epler, J. (2001) Identification manual for the larval Chironomidae (Diptera) of North and South Carolina. A guide to the taxonomy of the midges of the southeastern United States, including Florida. St. Johns River Water Management District, North Carolina Department of Environment and Natural Resources, State of North Carolina, 526 pp.
Escobar-Briones, E. & Alcocer, J. (2002) Caecidotea williamsi (Crustacea: Isopoda: Asellidae), a new species from a saline crater-lake in the eastern Mexican Plateau. Hydrobiologia, 477 (1–3), 93–105.
https://doi.org/10.1023/A:1021013132626
Filonov, A., Tereshchenko, I. & Alcocer, J. (2006) Dynamic response to mountain breeze circulation in Alchichica, a crater lake in Mexico. Geophysical Research Letters, 33 (7), L07404.
https://doi.org/10.1029/2006GL025901
Gunderina, L.I. (2012) Species-specific PCR primers for identification of the sibling species Chironomus plumosus (Linnaeus, 1758) and Chironomus balatonicus (Devai, Wülker et Scholl, 1983) (Chironomidae, Diptera). Proceedings of the 18th International Symposium on Chironomidae - Fauna norvegica, 31, 151–157.
https://doi.org/10.5324/fn.v31i0.1381
Guryev, V.P., Makarevitch, I., Blinov, A. & Martin, J. (2001) Phylogeny of the genus Chironomus (diptera) inferred from DNA sequences of mitochondrial cytochrome b and cytochrome oxidase I. Molecular Phylogenetics and Evolution, 19 (1), 9–21.
https://doi.org/10.1006/mpev.2001.0898
Guryev, V.P. & Blinov, A.G. (2002) Phylogenetic relationships among Holarctic populations of Chironomus entis and Chironomus plumosus in view of possible horisontal transfer of mitochondrial genes. Russian Journal of Genetics, 38 (3), 239–243
https://doi.org/10.1023/A:1014842415628
Hebert, P.D.N., Penton, E.H., Burns, J.M., Janzen, D.H. & Hallwachs, W. (2004) Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proceedings of the National Academy of Sciences of the United States of America, 101 (41), 14812–14817.
https://doi.org/10.1073/pnas.0406166101
Hebert, P. D.N., Ratnasingham, S., Zakharov, E.V., Telfer, A.C., Levesque-Beaudin, V., Milton, M.A., Pedersen, S., Jannetta, P. & deWaard, J.R. (2016) Counting animal species with DNA barcodes: Canadian insects. Philosophical Transactions of the Royal Society B Biological Sciences, 371, 20150333.
https://doi.org/10.1098/rstb.2015.0333
Hernández, M.C., Alcocer, J., Oseguera, L.A. & Escobar, E. (2014) Profundal benthic invertebrates in an oligotrophic tropical lake: different strategies for coping with anoxia. Journal of Limnology, 73 (2), 387–399.
https://doi.org/10.4081/jlimnol.2014.910Jabłońska-Barna, I., Szarek-Gwiazda, E. & Michailova, P. (2013) Environmental agents in Lake Łuknajno (Poland) affecting the genome of Chironomus melanotus Keyl, 1961 (Diptera, Chironomidae) — a new species of Polish fauna. Oceanological and Hydrobiological Studies, 42 (2), 164–172.
https://doi.org/10.2478/s13545-013-0070-2Jansson, A. (1979) A new species of Krizousacorixa from Mexico (Heteroptera, Corixidae). The Pan-Pacific Entomologist, 55, 258–260.
Johannsen, O.A. (1905) Aquatic nematocerous Diptera II. Chironomidae. In: Needham, I.G., Morton, K.I. & Johannsen, O.A. (Eds.), May flies and midges of New York. Third report on aquatic insects, 86. New York State Education Department, Albany, pp. 16–37.
https://doi.org/10.5962/bhl.title.20757
Johannsen, O.A. (1908) New North American Chironomidae. In: Felt, E.P. (Ed.), 23rd report of the State Entomologist on injurious and other insects of the State New York, 1907, 124. New York State Museum Bulletins, U.S.A., pp. 264 –285.
Kajak, Z. & Prus, P. (2003) Seasonal and year-to-year variation of numbers of Chironomus plumosus L. and Tubificidae in a lowland reservoir: Regularities, causes, mechanisms. Polish Journal of Ecology, 51 (3), 339–351.
Keyl, H.-G. (1962) Chromosomenevolution bei Chironomus II. Chromosomenumbauten und phylogenetische Beziehungen der Arten. Chromosoma, 13 (4), 464–514.
https://doi.org/10.1007/BF00327342
Kiknadze, I.I., Shilova, A., Kekris, I., Shobanov, N., Zelenzov, N., Grebenjuk, A., Istomina, A. & Praslov, B. (1991) Karyotype and morphology of larvae in Chironomini. Atlas. Novosibirsk, Nauka, 114 pp.
Kondo, N.I., Ueno, R., Ohbayashi, K., Golygina, V. & Takamura, K. (2016) DNA barcoding supports reclassification of Japanese Chironomus species (Diptera: Chironomidae). Entomological Science, 19 (4), 337–350.
https://doi.org/10.1111/ens.12212
Kumar, S., Stecher, G. & Tamura, K. (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33 (7), 1870–1874.
https://doi.org/10.1093/molbev/msw054
Lindeberg, B. & Wiederholm, T. (1979) Notes on the taxonomy of European species of Chironomus (Diptera: Chironomidae). Recent development in chironomid studies (Diptera: Chironomidae). Scandinavian Entomology Supplement. International Review of Hydrobiology, 10, 99–116.
Lindegaard, C. (1995) Chironomidae (Diptera) of European Cold Springs and Factors Influencing their Distribution. Journal of the Kansas Entomological Society, 68 (2), 108–131.
Martin, J. (1979) Chromosomes as tools in taxonomy and phylogeny of Chironomidae (Diptera). Entomologica Scandinavica Supplement, 10, 67–74.
Martin, J. (2011) From bands to base pairs: Problems in the identification of species using the example of Chironomus oppositus Walker (Honorary Thienemann Lecture). In: Wang, X. & Liu, W. (Eds.), Contemporary Chironomid Studies. Proceedings of the 17th International Symposium on Chironomidae, 6–9 July, 2009, Nankai University, China. Nankai University Press, Tianjin, pp. 126–143.
Martin, J. (2017) North American species of the genus Chironomus as recognized by larval cytology and morphology. Available from: http://www.chironomidae.net/Martin/NACytfiles/NAChiron.html (accessed 19 June 2017)
Martin, J., Guryev, V. & Blinov, A. (2002) Population variability in Chironomus (Camptochironomus) species (Diptera, Nematocera) with a Holarctic distribution: evidence of mitochondrial gene flow. Insect Molecular Biology, 11 (5), 387–397.
https://doi.org/10.1046/j.1365-2583.2002.00348.x
Martin, S., Proulx, I. & Hare, L. (2008) Explaining metal concentrations in sympatric Chironomus species. Limnology and Oceanography, 53 (2), 411–419.
https://doi.org/10.4319/lo.2008.53.2.0411
Michailova, P. (1989) The polytene chromosomes and their significance to the systematics of the family Chironomidae, Diptera. Acta Zoologica Fennica, 186, 1–107.
Michailova, P., IIkova, J. & White, K. (2016) Implications of Genome Alterations in Chironomus bernensis Klȍtzli (Diptera) for Assessment of Trace Metal Pollution in Two Bulgarian Rivers. River Research and Applications, 32 (5), 914–924.
https://doi.org/10.1002/rra.2927
Montiel-Martínez, A., Ciros-Pérez, J., Ortega-Mayagoitia, E. & Elías-Gutiérrez, M. (2008) Morphological, ecological, reproductive and molecular evidence for Leptodiaptomus garciai (Osorio-Tafall 1942) as a valid endemic species. Journal of Plankton Research, 30 (10), 1079–1093.
https://dx.doi.org/10.1093/plankt/fbn067
Oliva, M.G., Lugo, A., Alcocer, J. & Cantoral-Uriza, E.A. (2006) Cyclotella alchichicana sp. nov. from a saline Mexican lake. Diatom Research, 21 (1), 81–89.
https://doi.org/10.1080/0269249x.2006.9705653
Osorio-Tafall, B.F. (1942) Un nuevo Diaptomus del México Central (Copepoda, Diaptomidae). Revista Brasileira de Biologia, 2, 147–154.
Pedrosa, J.A.M., Cocchiararo, B., Bordalo, M.D., Rodrigues, A.C.M., Soares, A.M.V.M., Barata, C. & Nowak, C. (2017) The role of genetic diversity and past-history selection pressures in the susceptibility of Chironomus riparius populations to environmental stress. Science of the total Environment, 576 (15), 807–816.
https://doi.org/10.1016/j.scitotenv.2016.10.100
Paggi, A.C. (2009) Chironomidae. In: Domínguez, E. & Fernández, H. (Eds.), Macroinvertebrados bentónicos sudamericanos. Sistemática y biología. Fundación Miguel Lillo, Tucumán, pp. 383–409.
Pfenninger, M., Nowak, C., Kley, C., Steinke, D. & Streit, B. (2007) Utility of DNA taxonomy and barcoding for the inference of larval community structure in morphologically cryptic Chironomus (Diptera) species. Molecular Ecology, 16 (9), 1957–1968.
https://doi.org/10.1111/j.1365-294x.2006.03136.x
Pfrender, M.E., Hawkins, C.P., Bagley, M., Courtney, G.W., Creutzburg, B.R, Epler, J.H., Fend, S., Ferrington, L.C., Hartzell, P.L., Jackson, S., Larsen, D.P., Levesque, C.A., Morse, J.C., Petersen, M.J., Ruiter, D., Schindel, D. & Whiting, M. (2010) Assessing Macroinvertebrate Biodiversity in Freshwater Ecosystems: Advances and Challenges in DNA-Based Approaches. Quarterly Review of Biology, 85 (3), 319–340.
https://doi.org/10.1086/655118
Prat, N., Ribera, C., Rieradevall, M., Villamarin, C. & Acosta, R. (2013) Distribution, abundance and molecular analysis of Barbadocladius Cranston & Krosch (Diptera, Chironomidae) in tropical, high altitude Andean streams and rivers. Neotropical Entomology, 42 (6), 607–617.
https://doi.org/10.1007/s13744-013-0161-1
Proulx, I., Martin, J., Carew, M. & Hare, L. (2013) Using various lines of evidence to identify Chironomus species in eastern Canadian lakes. Zootaxa, 3741 (4), 401–458.
https://doi.org/10.11646/zootaxa.3741.4.1
Ramírez-García, P. & Novelo, A. (1984) La vegetación acuática vascular de seis lagos- cráter del estado Puebla, México. Boletín de la Sociedad Botánica de México, 46, 75–88.
Richardi, V.S., Vicentini, M., Rebechi, D., Fávaro, L.F & Navarro-Silva, M.A. (2015) Morpho-histological characterization of immature of the bioindicator midge Chironomus sancticaroli Strixino and Strixino (Diptera, Chironomidae). Revista Brasileira de Entomologia, 59 (3), 240–250.
https://doi.org/10.1016/j.rbe.2015.07.003
Sari, A., Duran, M., Sen, A. & Bardakci, F. (2015) Investigation of Chironomidae (Diptera) relationships using mitochondrial COI gene. Biochemical Systematics and Ecology, 59, 229–238.
https://doi.org/10.1016/j.bse.2015.01.005
Sharley, D.J., Pettigrove, V. & Parsons, Y.M. (2004) Molecular identification of Chironomus spp. (Diptera) for biomonitoring of aquatic ecosystems. Australian Journal of Entomology, 43 (4), 359–365.
https://doi.org/10.1111/j.1440-6055.2004.00417.x
Shapiro, J.A. (2002) Genome organization and reorganization in evolution: formatting for computation and function. In: Van Speybroack, L., Van de Vijver, G. & de Waele, D. (Eds.), From Epigenesis to Epigenetics: The Genome in Context, pp. 111–134.
https://doi.org/10.1111/j.1749-6632.2002.tb04915.xShobanov, N.A., Shilova, A.I. & Belyanina, S.I. (1996) Extent and content of the genus Chironomus Meig. (Diptera, Chironomidae): review of world fauna. In: Shobanov, N.A. & Zinchenko, T.D. (Eds.), Ecology, Evolution and Systematics of Chironomids. Inst. Biol. Inland Waters & Inst. Ecol. Volga Basin, Russian Academy of Sciences, Tolyatti, Boruk, pp. 44–96.
Silvestro, D. & Michalak, I. (2012) RaxmlGUI: A graphical front-end for RAxML. Organisms Diversity and Evolution, 12 (4), 335–337.
https://doi.org/10.1007/s13127-011-0056-0
Stamatakis, A. (2006) RAxML-VI-HPC: Maximum likelihood-based phylogenetic analices with thousands of taxa and mixed models. Bioinformatics, 22 (21), 2688–2690.
https://doi.org/10.1093/bioinformatics/btl446
Strenzke, K. (1959) Revision der Gattung Chironomus Meig. I. Die Imagines von 15 norddeutschen Arten und Unterarten. Archiv für Hydrobiologie, 56, 1–42.
Suárez-Morales, E., Barrera-Moreno, O. & Ciros-Pérez, J. (2013) A new species of Cletocamptus Schmankewitsch, 1875 (Crustacea, Copepoda, Harpacticoida) from a high altitude saline lake in Central Mexico. Journal of Limnology, 72 (2), 313–325.
https://doi.org/10.4081/jlimnol.2013.e25
Townes, H.K. (1945) The Neartic species of the Tendipedini [Diptera, Tendipedidae (= Chironomidae)]. The American Midland Naturalist, 34, 1–206.
https://doi.org/10.2307/2421112Vallenduuk, H.J. & Moller Pillot, H.K.M. (1997) Key to the larvae of Chironomus in Western Europe. RIZA Rapport 97.053, 1997, pp. 1–13, appendices.
Vilaclara, G., Chávez, M., Lugo, A., González, H. & Gaytán, M. (1993) Comparative description of crater-lakes basic chemistry in Puebla State, Mexico. Verhandlungen des Internationalen Verein Limnologie, 25, 435–440.
Webb, C.J. & Scholl, A. (1985) Identification of larvae of European species of Chironomus Meigen (Diptera: Chironomidae) by morphological characters. Systematic Entomology, 10 (3), 353–372.
https://doi.org/10.1111/j.1365-3113.1985.tb00143.x
Wülker, W. (1973) Revision der Gattung Chironomus Mg. III. Europäische Arten des thummi Komplexes. Archiv für Hydrobiologie, 72 (3), 356 – 374.
Wülker, W., Devai, Gy. & Devai, I. (1989) Computer assisted studies of chromosome evolution in the genus Chironomus (Dipt.). Comparative and integrated analysis of chromosome arms A, E and F. Acta biologica Debrecina. Supplementum oecologica Hungarica, 2, 373–387.
Wülker, W. & Martin, J. (1974) A Review of the genus Chironomus (Diptera, Chironomidae). VI. Cytology of the maturus – complex. Studies in Natural Sciences, 1 (9), 1–21.
Wülker, W., Sublette, J.E. & Martin, J. (1991) Chironomus utahensis Malloch and Chironomus harpi new species and their karyosystemic relationships to other species of the decorus-group of Chironomus. Spixiana, 14, 71–94.
Wülker, W., Martin, J., Kiknadze. I.I., Sublette, J.E. & Michiels, S. (2009) Chironomus blaylocki n. sp. and C. bifurcatus n. sp., North American species near the base of the decorus-group. Zootaxa, 2023, 28–46.
Wülker, W., Kiknadze, I.I. & Istomina, A. (2011) Karyotypes of Chironomus Meigen (Diptera: Chironomidae) species from Africa. Comparative Cytogenetics, 5 (1) 23–46.
https://doi.org/10.3897/compcytogen.v5i1.975