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Discrimination between four Simocephalus species from Slovakia using a PCR-RFLP technique

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Abstract

Planktonic crustaceans are traditionally identified based on morphological and morphometric characters. However, such characters may be hardly distinguishable and often overlap between species. A probability of misidentification is thus relatively high. Molecular techniques may increase the accuracy of identification if appropriate markers are used. Aim of our work was to develop a simple molecular procedure enabling discrimination between four species of Simocephalus occurring in Europe. PCR-RFLP technique proved to be suitable for such discrimination. Within the 709 bp fragment of mitochondrial cytochrome c oxidase subunit 1 gene we found unique combinations of restriction sites of the BbsI and SacI enzymes for Simocephalus vetulus, S. exspinosus, S. serrulatus and S. congener. PCR products of samples from several locations in Slovakia were digested with the two enzymes and electrophoresed on an agarose gel. The restriction patterns were clearly visible and easily distinguishable. This method is applicable for identifying the four species in any life-stage. Considering its simplicity and cost-effectiveness it can be widely used as a diagnostic tool for discriminating between Simocephalus species with overlapping morphologic characters.

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References

  • Ahmed A.O.A., Mukhtar M.M., Kools-Sijmons M., Fahal A.H., de Hoog S., van den Ende G.B., Zijlstra E.E., Verbrugh H., Abughroun E.S.A.M., Elhassan A.M. & van Belkum A. 1999. Development of a species-specific PCR-restriction fragment length polymorphism analysis procedure for identification of Madurella mycetomatis. J. Clin. Microbiol. 37: 3175–3178. PMID: 85521

    PubMed  CAS  Google Scholar 

  • Folmer O., Black M., Hoeh W., Lutz R. & Vrijenhoek R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol. Mar. Biol. Biotech. 3: 294–299. PMID: 7881515

    CAS  Google Scholar 

  • Gajardo G., Crespo J., Triantafyllidis A., Tzika A., Baxevanis A.D., Kappas I. & Abatzopoulos T.J. 2004. Species identi-fication of Chilean Artemia populations based on mitochondrial DNA RFLP analysis. J. Biogeogr. 31: 547–555. DOI: 10.1111/j.1365-2699.2003.01046.x

    Article  Google Scholar 

  • Galan M., Pagès M. & Cosson J.-F. 2012. Next-generation sequencing for rodent barcoding: species identification from fresh, degraded and environmental samples. PLoS ONE 7(11): e48374. DOI: 10.1371/journal.pone.0048374

    Article  PubMed  CAS  Google Scholar 

  • Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. 41: 95–98.

    CAS  Google Scholar 

  • Hann B.J. 1987. Naturally occurring interspecific hybridization in Simocephalus (Cladocera, Daphniidae): its potential significance. Hydrobiologia 145: 219–224. DOI: 10.1007/BF02530283

    Article  Google Scholar 

  • Hann B.J. 1995. Genetic variation in Simocephalus (Anomopoda: Daphniidae) in North America: patterns and consequences. Hydrobiologia 307: 9–14. DOI: 10.1007/BF00031992

    Article  Google Scholar 

  • Hebert P.D.N. 1985. Interspecific hybridization between cyclic parthenogens. Evolution 39: 216–220. DOI: 10.2307/2408534

    Article  Google Scholar 

  • Hebert P.D.N., Cywinska A., Ball S.L. & deWaard J.R. 2003a. Biological identifications through DNA barcodes. Proc. R. Soc. Lond. B 270: 313–322. DOI: 10.1098/rspb.2002.2218

    Article  CAS  Google Scholar 

  • Hebert P.D.N., Ratnasingham S. & deWaard J.R. 2003b. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc. R. Soc. Lond. B 270(Suppl. 1): S96–S99. DOI: 10.1098/rsbl.2003.0025

    Article  CAS  Google Scholar 

  • Hudec I. 1993. Notes to the distribution of the genus Simocephalus (Crustacea: Daphniiformes, Daphniidae) in Slovakia. Biologia 48: 141–147.

    Google Scholar 

  • Hudec I. 1995. Variability of Simocephalus vetulus (Crustacea: Anomopoda, Daphniidae) in Slovakia. Biologia 50: 465–473.

    Google Scholar 

  • Ingaki Y., Ehara M., Watanabe K.I., Hayashi-Ishimaru Y. & Ohama, T. 1998. Directionally evolving genetic code: the UGA codon from stop to tryptophan in mitochondria. J. Mol. Evol. 47: 378–384. PMID: 9767683

    Article  Google Scholar 

  • Johns G.C. & Avise J.C. 1998. A comparative summary of genetic distances in the vertebrates from the mitochondrial cytochrome b gene. Mol. Biol. Evol. 15: 1481–1490. PMID: 12572611

    Article  PubMed  CAS  Google Scholar 

  • Kalous L., ŠlechtovČech M. 2010. Do small fish mean no voucher? Using a flatbed desktop scanner to document larval and small specimens before destructive analyses. J. Appl. Ichthyol. 26: 614–617. DOI: 10.1111/j.1439-0426.2010.01471.x

    Article  Google Scholar 

  • Kohout J., Pekárik L., Šedivá A., Didenko A., Čiampor F. & Čiamporová-Zaťovičová Z. 2013. Discrimination between invasive Ponto-Caspian gobies using a PCR-RFLP method. J. Appl. Ichthyol., published online. DOI: 10.1111/jai.12315

    Google Scholar 

  • Kress W.J., Wurdack K.J., Zimmer E.A., Weigt L.A. & Janzen D.H. 2005. Use of DNA barcodes to identify flowering plants. Proc. Natl. Acad. Sci. U.S.A. 102(23): 8369–8374. DOI: 10.1073/pnas.0503123102

    Article  PubMed  CAS  Google Scholar 

  • Mirhendi H., Makimura K., Khoramizadeh M. & Yamaguchi H. 2006. A one-enzyme PCR-RFLP assay for identification of six medically important Candida species. Jpn. J. Med. Mycol. 47: 225–229. PMID: 16940958

    Article  CAS  Google Scholar 

  • Orlova-Bienkowskaja M.Y. 2001. Cladocera: Anomopoda: Daphniidae: genus Simocephalus. In: Dumont H.J.F. (ed.), Guides to the Identification of the Microinvertebrates of the Continental Waters of the World 17, SBA Acad. Publ. Hague, 130 pp. ISBN-10: 9057820900

    Google Scholar 

  • Schwenk K. 1993. Interspecific hybridization in Daphnia: Distinction and origin of hybrid matrilines. Mol. Biol. Evol. 10: 1289–1302. PMID: 8277855

    PubMed  CAS  Google Scholar 

  • Širca S., Geric Stare B., Strajnar P. & Urek G. 2010. PCR-RFLP diagnostic method for identifying Globodera species in Slovenia. Phytopathol. Mediterr. 49: 361–369.

    Google Scholar 

  • Šrámek-Hušek R. 1962. Cladocera — perloočky, pp. 174–410. In: Šrámek-Hušek R., Straškraba M. & Brtek J. (eds), Lupenonožci — Branchiopoda, Fauna ČSSR 16, NČSAV, Praha.

    Google Scholar 

  • Traub R.J., Robertson I.D., Irwin P., Mencke N. & Thompson A. 2004. Application of a species-specific PCR-RFLP to identify Ancylostoma eggs directly from canine faeces. Vet. Parasitol. 123: 245–255. DOI: 10.1016/j.vetpar.2004.05.026

    Article  PubMed  CAS  Google Scholar 

  • Young Sh-S., Ni M.-H. & Liu M.-Y. 2012. Systematic study of the Simocephalus sensu stricto species group (Cladocera: Daphniidae) from Taiwan by morphometric and molecular analyses. Zool. Stud. 51(2): 222–231.

    Google Scholar 

  • Zapata M.A., Cienfuegos A.V., Quirós O.I., Quińones M.L., Luckhaart S. & Correa M.M. 2007. Discrimination of seven Anopheles species from San Pedro de Urabá, Antioquia, Colombia, by polymerase chain reaction-restriction fragment length polymorphism analysis of its sequences. Am. J. Trop. Med. Hyg. 77(1): 67–72. PMID: 17620632

    PubMed  CAS  Google Scholar 

  • Zaret T. 1969. Predation-balanced polymorphism of Ceriodaphnia cornuta Sars. Limnol. Oceanogr. 14: 301–303. DOI: 10.4319/lo.1969.14.2.0301

    Article  Google Scholar 

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Kohout, J., Illyová, M., Čiampor, F. et al. Discrimination between four Simocephalus species from Slovakia using a PCR-RFLP technique. Biologia 69, 76–79 (2014). https://doi.org/10.2478/s11756-013-0285-0

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