PCR-based molecular markers for identification of taxa from the Calypogeia fissa complex (Jungermanniopsida, Calypogeiaceae)

Open access


Within Calypogeia fissa, two subspecies connected with geographic distribution are formally recognized: C. fissa subsp.fissa in Europe and C. fissa subsp.neogea in North America. Isoenzyme studies have shown that the European subspecies is genetically differentiated and composed of three genetically distinct groups PS, PB and G. The PS group has the most distinctive morphological features, but no morphological diagnostic traits have been found for groups PB and G. The sequence characterized amplified region (SCAR) markers developed on the basis of ISSR markers, applied in the study, allowed the delimitation of all groups distinguished in Europe within the C. fissa complex (PS, PB and G). The markers also revealed genetic differences between the European and American subspecies. Five primer pairs (Cal01, Cal03-Cal06) of the six pairs studied are useful as the diagnostic tool for the identification of particular groups from the C.fissa complex. The examined SCAR markers showed that the PS group of C.fissa subsp.fissa was the most distinct; it differed from both groups PB and G as well as from C.fissa subsp.neogea. All plants determined on the basis of diagnostic isozyme loci as the PS group amplified a longer product (380 bp) of the Cal04 primer pair than the rest of studied groups and yielded no amplification products in Cal03, Cal05 and Cal06 primers. The primer pair Cal03 distinguished the plants of the PB group from the remaining groups, since only the PB group generated a PCR product of about 290 bp. The genetic differences between all four studied groups of the C.fissa complex were supported by DNA sequences of the SCAR marker Cal04.

Anuniwat A., Chaveerach A., Tance T. & Sudmoon R. 2009. Development od SCAR markers for species identification of the genus Nepenthes (Nepenthaceae). Pakistan J. Biol. Sci.12: 1455-1461.

Bailey C. D., Hughes C. E. & Harris S. A. 2004. Using RAPDs to identify DNA sequence loci for species level phylogeny reconstruction: an example from Leucaena (Fabaceae). Syst. Bot. 29: 4-14.

Basha S. D & Sujatha M. 2007. Inter and intra-population variability of Jatropha curcas (L.) characterized by RAPD and ISSR markers and development of population- specific SCAR markers. Euphytica 156:375-386.

Bączkiewicz A., Sawicki J., Buczkowska K., Polok K. & Zieliński R. 2008. Application of different DNA markers in studies on cryptic species of Aneura pinguis (Jungermanniopsida, Metzgeriales). Cryptogam. Bryol. 29: 3-21.

Boiss elier-Duba yle M. C., Jubier M. F., Lejeune B. & Bischler H. 1995. Genetic variability in the three subspecies of Marchantia polymorpha (Hepaticae): isozymes RFLP and RAPD markers. Taxon 44: 363-376.

Buczkowska K. 2004. Genetic differentiation of Calypogeiafissa (L.) Raddi (Hepaticae, Jungermanniales) in Poland. Plant Syst. Evol. 247: 187-201. DOI 10.1007/ s00606-003-0156-9

Buczkowska K. 2010. Morphological differentiation of Calypogeiamuelleriana (Jungermanniales, Hepaticae) in Poland. Biodiv. Res. Conserv. 17: 23-32. DOI 10.2478/v10119-010-0004-4

Buczkowska K. & Bączkiewicz A. 2011. New taxon of the genus Calypogeia (Jungermanniales, Hepaticae) in Poland. Acta Soc. Bot. Pol. 80(4): 327-333. DOI 10.5586/asbp.2011.039

Buczkowska K. & Dab ert M. 2011. The development of species-specific SCAR markers for delimitation of Calypogeia species. J. Bryol. 33: 291-299.

Buczkowska K., Sawicki J., Szczecińska M., Klama H. & Bączkiewicz A. 2012a. Allopoliploid speciation of Calypogeia sphagnicola (Jungermanniopsiada, Calypogeiaceae) based on isozyme and DNA markers. Plant Syst. Evol. 298: 549-560.

Buczkowska K., Sawicki J., Szczecińska M., Klama H. & Bączkiewicz A. 2012b. Isozyme and DNA markers reveal a new genetically distinct taxon of Calypogeiasphagnicola (Jungermanniopsida, Calypogeiaceae). Pol. Bot. J. 57: 95-107.

Buczkowska K., Sawicki J., Szczecińska M., Rosadziński S., Rabska M., Bączkiewicz A. 2011. Two morphologically distinct groups of the Calypogeia fissa complex were found in Europe. Biodiv. Res. Conserv. 23: 7-19. DOI 10.2478/v10119-011-0014-x Dams holt K. 2002. Illustrated Flora of Nordic Liverworts and Hornworts. 837 pp. Nordic Bryological Society, Lund.

Das M., Bhattacharya S. & Pal A. 2005. Generation and characterization of SCARs by cloning and sequencing of RAPD products: A strategy for species-specific marker development in bamboo. Ann. Bot. 95: 835-841.

Fuselier L., Davison P. G., Clements M., Shaw B., Devos N., Heinrichs J., Hentschel J., Sab ovlj evic M., Szovenyi P., Schuette S., Hofba uer W & Shaw A. J. 2009. Phylogeographic analysis reveal distinct lineages of the liverworts Metzgeria furcata (L.) Dumort. and Metzgeriaconjugata Lindb. (Metzgeriaceae) in Europe and North America. Biol. J. Linn. Soc. 98: 745-756.

Gao L., Chen W. Q. & Liu T. G. 2010. Development of a SCAR marker by inter-simple sequence repeat for diagnosis of dwarf bunt of wheat and detection of Tiletacontroversa Kühn. Folia Microbiol. 55: 258-264.

Hassel K. & Gunnarsson U. 2003. The use of inter simple sequence repeats (ISSR) in bryophyte population studies. Lindbergia 28: 152-157.

Hedenäs L., Bisa ng I., Korpela inen H & Cronholm B. 2010. The true sex ratio in European Pseudocalliergontrifarium (Bryophyta: Amblystegiaceae) revealed by a novel molecular approach. Biol. J. Linn. Soc. 100: 132-140.

Heinrichs J., Hentschel J., Bombosch A., Fiebig A., Reise J., Edelmann M., Kreier H. P., Schäfer-Verwimp A., Caspa ri S., Schmidt A. R., Zhu R. L., Konrat M., Shaw B. & Shaw A. J. 2010. One species or at least eight? Delimitation and distribution of Frullaniatamarisci (L.) Dumort. s. l. (Jungermanniopsida, Porellales) inferred from nuclear and chloroplast DNA markers. Mol. Phylogenet. Evol. 56: 1105-1114.

Jankowiak K., Buczkowska K. & Szweykowska-Kulińska Z. 2005. Successful extraction of DNA from 100-yearold herbarium specimens of the liverwort Bazzaniatrilobata. Taxon 54: 335-336.

Koła W. 1989. Analiza florystyczno-ekologiczna wątrobowców Wzgórz Trzebnicko-Ostrzeszowskich. In: I. Kuczyńska (ed.). Zmiany szaty roślinnej Dolnego Śląska pod wpływem działalności człowieka. Acta Univ. Wratislaviensis 1156, Prace Botaniczne 44: 305-355.

Korpelainen H., Bisang I., Hedenäs L. & Kolehmainen J. 2008. The first sex-specific molecular marker discoverd in the moss Pseudocalliergon trifarium. J. Hered. 99: 581-587.

Kreier H. P., Feldberg K., Mahr F., Bombosch A., Schmidt A. R., Zhu R. L., Konrat M., Shaw B., Shaw A. J. & Heinrichs J. 2010. Phylogeny of the leafy liverwort Ptilidium: Cryptic speciation and shared haplotypes between Northern and Southern Hemispheres. Mol. Phylogenet. Evol. 57: 1260-1267. Lister D. L., Bower M. A., Howe C. J. & Jones M. K. 2008. Extraction and amplification of nuclear DNA from herbarium specimens of emmer wheat: a method for assessing DNA preservation by maximum amplicon length recovery. Taxon 57: 254-258.

Liu Y., Yan H. F., Cao T. & Ge X. J. 2010. Evaluation of 10 plant barcodes in bryophyte (Mosses). J. Syst. Evol. 48: 36-46.

Mehes M. S., Nkongolo K. K. & Michael P. 2007. Genetic analysis of Pinus strobus and Pinus monticola populations from Canada using ISSR and RAPD markers: development of genome-specific SCAR markers. Plant Syst. Evol. 267: 47-63.

Melotto M., Afanad or L. & Kell y J. D. 1996. Development of a SCAR marker linked to the I gene in common bean. Genome 39: 1216-1219.

Murray M. & Thomps on W. F. 1980. Rapid isolation of molecular weight plant DNA. Nucleic Acids Res. 8: 4321-4325.

Müller K. 1951-1958. Die Lebermoose Europas In: Dr L Rabenhorst’s Kryptogamen Flora von Deutschland …sterreich und der Schweiz. 3rd ed. Akademische Verlagsgesellaschaft Geest & Portig K.-G., Leipzig.

Pacak A. & Szweykowska-Kulińska Z. 2003.Organellar inheritance in liverworts. An example of Pellia borealis. J. Mol. Evol. 56: 11-17.

Paran I. & Michelm ore R. W. 1993. Development of reliable PCR based morker linked to downy mildew resistence gene in lettuce. Theor. Appl. Genet. 85: 985-993.

Paton J. 1999. The Livervort Flora of the British Isles. 626 pp. Harley Books, Colchester.

Sawicki J. & Szczecińska M. 2011. A comparison of PCRbased markers for the molecular identification of Sphagnum species of the section Acutifolia. Acta Soc. Bot. Pol. 80(3): 185-192.

Schuster R. M. 1969. The Hepaticae and Anthocerotae of North America east of the hundredth meridian, vol. 2, 1062 pp. Columbia University Press, New York- London.

Shaw A. J., Boles S. B. & Shaw B. 2008. A phylogenetic delimitation of the “Sphagnum subsecundum complex” (Sphagnaceae, Bryophyta). Am. J. Bot. 95: 731-744.

Shaw A. J., Cox C. J. & Boles S. B. 2003. Polarity of peatmoss (Sphagnum) evolution: who says bryophytes have no roots? Am. J. Bot. 90: 1777-1787.

Shaw A. J., Melosik I. & Cox C. J. 2005. Divergent and reticulate evolution in closely related species of Sphagnum section Subsecunda. Bryologist 108: 363-376.

Stech M. & Quandt D. 2010. 20,000 species and five key markers: The status of molecular bryophyte phylogenetic. Phytotaxa 9: 196-228.

Su H., Wang L., Liu L., Chi X. & Zhang Y. 2008. Use of inter-simple sequence repeat markers to develop strain-specific SCAR markers for Flammulina velutipes. J. Appl. Genet. 49: 233-235.

Szweykowska-Kulińska Z., Pacak A. & Jankowiak K. 2002. New DNA markers for discrimination between closely-related species and for the reconstruction of historical events; an example using liverworts. Cell. Mol. Biol. Lett. 7: 403-416.

Szweykowski J. 2006. An annotated checklist of Polish liverworts. In: Z. Mirek (ed.). Biodiversity of Poland, 4, 114 pp. W. Szafer Institute of Botany, Polish Academy of Sciences, Kraków.

Tamura K., Dudley J., Nei M. & Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.1. Mol. Biol. Evol. 24: 1596-1599.

Vanderpoorten A., Hedenäs L. & Jacquema rt A.L. 2003. Differentiation in DNA fingerprinting and morphologyamong species of the pleurocarpous moss genus Rhytidiadelphus (Hylocomniaceae). Taxon 52: 229-236.

Ye Q., Qiu Y., Quo Y., Chen J., Yang S., Zhao M & Fu C. 2006. Species-specific SCAR marker for authentication of Sinocalycanthus chinensis. J. Zhejiang Univ. Sci. B. 7: 868-872.

Biodiversity Research and Conservation

The Journal of Adam Mickiewicz University

Journal Information


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 103 103 3
PDF Downloads 44 44 1