Skip to main content
SpringerLink
Log in

Ploidy level and genome size variation in the homosporous ferns Botrychium s.l. (Ophioglossaceae)

  • Original Article
  • Published:
Plant Systematics and Evolution Aims and scope Submit manuscript

Abstract

Recent cytological and molecular studies have investigated genome size variation and evolution in the homosporous ferns, but representatives of the Ophioglossaceae were largely overlooked, despite their evolutionary singularity. Flow cytometry analysis was performed on 41 individuals of eight species of the genera Botrychium (B.), Botrypus, and Sceptridium to estimate their ploidy level variation. In a subset of individuals, we also estimated the absolute genome size and corresponding C values. Additionally, a classical chromosome count was made on the recently described species B. alaskense. Ploidy level and new genome size records were determined for Botrychium alaskense, B. boreale, B. lanceolatum, B. “neolunaria” ined., B. pinnatum, Botrypus virginianus and Sceptridium multifidum. In addition, we confirmed the genome size of B. matricariifolium, B. minganense and B. lunaria. Two of the three major sub-clades of Botrychium differ slightly in their averaged homoploid genome size (subclade Lanceolatum, 24.72 ± 0.40 pg; subclade Lunaria, 27.51 ± 0.47 pg). Flow cytometry and chromosome counting confirmed that B. alaskense is a tetraploid. A new hexaploid cytotype, putatively formed through an autopolyploidization from the sympatric tetraploid cytotype, was detected in a single individual of B. boreale. This is only the second report of hexaploidy in the genus Botrychium and our data highlight the potential to find other ploidy levels within other Botrychium species. Interestingly, no difference within the monoploid genome sizes was detected between ploidy levels, thus supporting the hypothesis of genome size stability after polyploidization and rejecting the scenario of genome downsizing.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Bai C, Alverson WS, Follansbee A, Waller DM (2012) New reports of nuclear DNA content for 407 vascular plant taxa from the US. Ann Bot (Oxford) 110:1623–1629. doi:10.1093/aob/mcs222

    Article  CAS  Google Scholar 

  • Bainard JD, Henry T, Bainard LD, Newmaster SG (2011) DNA content variation in monilophytes and lycophytes: large genomes that are not endopolyploid. Chromosom Res 19:763–775. doi:10.1007/s10577-011-9228-1

    Article  CAS  Google Scholar 

  • Barker MS (2013) Karyotype and genome evolution in pteridophytes. In: Leitch IJ, Greilhuber J, Doležel J, Wendel JF (eds) Plant genome Divers, vol 2. Wien: Springer, pp 245–253. Phys Struct Behav Evol plant genomes. doi:10.1007/978-3-7091-1160-4

  • Brandes A, Heslop-Harrison JS, Kamm A, Kubis S, Doudrick RL, Schmidt T (1997) Comparative analysis of the chromosomal and genomic organization of Ty1-copia-like retrotransposons in pteridophytes, gymnosperms and angiosperms. Pl Molec Biol 33:11–21. doi:10.1023/A:1005797222148

    Article  CAS  Google Scholar 

  • Casson JI, Shackleford I, Parker I, Schultz J (2002) Conservation approach for goblin fern (Botrychium mormo W.H. Wagner). USDA Forest Service, Eastern Region, Milwaukee

    Google Scholar 

  • Clausen RT (1936) A monograph of the Ophioglossaceae. Mem Torrey Bot Club 19:1–177

    Google Scholar 

  • Dalgaard V (1989) Additional chromosome numbers in vascular plants from the Disko Bugt area (west Greenland). Willdenowia 19:199–213

    Google Scholar 

  • Dauphin B, Vieu J, Grant JR (2014) Molecular phylogenetics supports widespread cryptic species in moonworts (Botrychium s.s., Ophioglossaceae). Amer J Bot 101:128–140. doi:10.3732/ajb.1300154

    Article  Google Scholar 

  • Development Core Team R (2015) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna

    Google Scholar 

  • Doležel J, Sgorbati S, Lucretti S (1992) Comparison of three DNA fluorochromes for flow cytometric estimation of nuclear DNA content in plants. Physiol Pl (Copenhagen) 85:625–631. doi:10.1111/j.1399-3054.1992.tb04764.x

    Article  Google Scholar 

  • Doležel J, Greilhuber J, Lucretti S, Meister A, Lysák MA, Nardi L, Obermayer R (1998) Plant genome size estimation by flow cytometry: inter-laboratory comparison. Ann Bot (Oxford) 82:17–26

    Article  Google Scholar 

  • Doležel J, Greilhuber J, Suda J (2007) Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc 2:2233–2244. doi:10.1038/nprot.2007.310

    Article  PubMed  Google Scholar 

  • Fabbri F (1963) Primo supplement alle Tavole Cromosomiche delle Pteridophyta di Alberto Chiarugi. Caryologia 16:237–335

    Article  Google Scholar 

  • Farrar DR (2011) Systematics and taxonomy of genus Botrychium, pp 1–29. Available at: http://www.public.iastate.edu/~herbarium/botrychium.html

  • Gagnidze RI, Tsn G, Danelia IM, Churadze MV (1998) Chromosome numbers of the species of the Georgian flora. Bot Zhurn (Moscow & Leningrad) 83:143–147

    Google Scholar 

  • Greilhuber J, Doležel J, Lysák MA, Bennett MD (2005) The origin, evolution and proposed stabilization of the terms ‘genome size’ and ‘C value’ to describe nuclear DNA contents. Ann Bot (Oxford) 95:255–260. doi:10.1093/aob/mci019

    Article  CAS  Google Scholar 

  • Hauk WD, Haufler CH (1999) Isozyme variability among cryptic species of Botrychium subgenus Botrychium (Ophioglossaceae). Amer J Bot 86(5):614–633

    Article  CAS  Google Scholar 

  • Hauk WD, Parks CR, Chase MW (2003) Phylogenetic studies of Ophioglossaceae: evidence from rbcL and trnL-F plastid DNA sequences and morphology. Molec Phylogen Evol 28:131–151. doi:10.1016/S1055-7903(03)00032-0

    Article  CAS  Google Scholar 

  • Hauk WD, Kennedy L, Hawke HM (2012) A phylogenetic investigation of Botrychium ss (Ophioglossaceae): evidence from three plastid DNA sequence datasets. Syst Bot 37:320–330. doi:10.1600/036364412X635377

    Article  Google Scholar 

  • Henry TA, Bainard JD, Newmaster SG (2014) Genome size evolution in Ontario ferns (Polypodiidae): evolutionary correlations with cell size, spore size, and habitat type and an absence of genome downsizing. Genome 57:555–566. doi:10.1139/gen-2014-0090

    Article  CAS  PubMed  Google Scholar 

  • Hultén E (1968) Flora of Alaska and neighboring territories—a manual of the vascular plants. Stanford University Press, Stanford

    Google Scholar 

  • Kato M (1987) A phylogenetic classification of Ophioglossaceae. Gard Bull Singapore 40:1–14

    Google Scholar 

  • Kovács GM, Balázs T, Pénzes Z (2007) Molecular study of arbuscular mycorrhizal fungi colonizing the sporophyte of the eusporangiate rattlesnake fern (Botrychium virginianum, Ophioglossaceae). Mycorrhiza 17:597–605. doi:10.1007/s00572-007-0137-2

    Article  PubMed  Google Scholar 

  • Leitch IJ, Bennett MD (2004) Genome downsize in polyploids plants. Biol J Linn Soc 82:651–663. doi:10.1111/j.1095-8312.2004.00349.x

    Article  Google Scholar 

  • Leitch IJ, Soltis DE, Soltis PS, Bennett MD (2005) Evolution of DNA amounts across land plants (Embryophyta). Ann Bot (Oxford) 95:207–217. doi:10.1093/aob/mci014

    Article  CAS  Google Scholar 

  • Loureiro J, Rodriguez E, Doležel J, Santos C (2007) Two new nuclear isolation buffers for plant DNA flow cytometry: a test with 37 species. Ann Bot (Oxford) 100:875–888. doi:10.1093/aob/mcm152

    Article  CAS  Google Scholar 

  • Löve A, Löve D (1976) IOPB chromosome number reports LIII. Taxon 25:483–500

    Google Scholar 

  • Paris CA, Wagner FS, Wagner WH (1989) Cryptic species, species delimitation, and taxonomic practice in the homosporous ferns. Amer Fern J 79:46–54. doi:10.2307/1547159

    Article  Google Scholar 

  • Peruzzi L, Cesca G, Puntillo D (2003) Isoëtes (Isoetaceae), Ophioglossum and Botrychium (Ophioglossaceae) in Calabria (S Italy): more karyological and taxonomical data. Caryologia 56:359–363. doi:10.1080/00087114.2003.10589345

    Article  Google Scholar 

  • Pryer KM, Schuettpelz E, Wolf PG, Schneider H, Smith AR, Cranfill R (2004) Phylogeny and evolution of ferns (monilophytes) with a focus on the early leptosporangiate divergences. Amer J Bot 91:1582–1598. doi:10.3732/ajb.91.10.1582

    Article  CAS  Google Scholar 

  • Ramsey J, Schemske DW (1998) Pathways, mechanisms, and rates of polyploid formation in flowering plants. Annual Rev Ecol Syst 29:467–501

    Article  Google Scholar 

  • Sharma AK, Sharma A (1980) Chromosome techniques—theory and practice. Butterworth-Heinemann, Oxford

    Google Scholar 

  • Shinohara W, Nakato N, Yatabe-Kakugawa Y, Oka T, Kim JK, Murakami N, Noda H, Sahashi N (2013) The use of matK in Ophioglossaceae phylogeny and the determination of Mankyua chromosome number shed light on chromosome number evolution in Ophioglossaceae. Syst Bot 38:564–570. doi:10.1600/036364413X670232

    Article  Google Scholar 

  • Soltis DE, Soltis PS, Schemske DW, Hancock JF, Thompson JN, Husband BC, Judd WS (2007) Autopolyploidy in angiosperms: have we grossly underestimated the number of species? Taxon 56:13–30. doi:10.2307/25065732

    Google Scholar 

  • Stensvold MC (2008) A taxonomic and phylogeographic study of the Botrychium lunaria complex. PhD Thesis, University of Iowa, Iowa

  • Wagner WH (1955) Cytotaxonomic observations on North American ferns. Rhodora 57:219–240

    Google Scholar 

  • Wagner FS (1993) Chromosomes of North American grapeferns and moonworts (Ophioglossaceae: Botrychium). Contr Univ Michigan Herb 19:83–92

    Google Scholar 

  • Wagner WH, Grant JR (2002) Botrychium alaskense, a new moonwort from the interior of Alaska. Amer Fern J 92:164–170. doi:10.1640/0002-8444(2002)

    Article  Google Scholar 

  • Wagner WH, Lord LP (1956) The morphological and cytological distinctness of Botrychium minganense and B. lunaria in Michigan. Bull Torrey Bot Club 83:261–280

    Article  Google Scholar 

  • Wagner WH, Wagner FS (1986) Three new species of moonworts (Botrychium subg. Botrychium) endemic in western North America. Amer Fern J 76:33–47. doi:10.2307/1547557

    Article  Google Scholar 

  • Wagner WH, Wagner FS (1990) Notes on the fan-leaflet group of moonworts in North America with descriptions of two new members. Amer Fern J 80:73–81. doi:10.2307/1547172

    Article  Google Scholar 

  • William JC (1996) Flora of the Yukon Territory. NRC Research Press, Ottawa

    Google Scholar 

  • Williams EW, Waller DM (2012) Phylogenetic placement of species within the genus Botrychium s.s. (Ophioglossaceae) on the basis of plastid sequences, amplified fragment length polymorphisms, and flow cytometry. Int J Pl Sci 173:516–531. doi:10.1086/664711

    Article  CAS  Google Scholar 

  • Winther JL, Friedman WE (2008) Arbuscular mycorrhizal associations in Lycopodiaceae. New Phytol 177:790–801. doi:10.1111/j.1469-8137.2007.02276.x

    Article  CAS  PubMed  Google Scholar 

  • Wood TE, Takebayashi N, Barker MS, Mayrose I, Greenspoon PB, Rieseberg LH (2009) The frequency of polyploid speciation in vascular plants. Proc Natl Acad Sci USA 106:13875–13879. doi:10.1073/pnas.0811575106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhukova PG, Petrovsky VV (1976) Chromosome numbers in some plant species from West Chukotka. II. Bot Zhurn 61:963–969

    Google Scholar 

  • Zika PF, Farrar DR (2009) Botrychium ascendens WH Wagner (Ophioglossaceae) in Newfoundland and notes on its origin. Amer Fern J 99:249–259. doi:10.1640/0002-8444-99.4.249

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the Fonds Marguerite Wüthrich and A. Matthey-Dupraz at the Université de Neuchâtel, the Fonds Dr. Joachim de Giacomi of the Swiss Academy of Natural Sciences, the Bourse de voyages of the Swiss Academy of Natural Sciences, and the Fonds de donations of the Université de Neuchâtel. The authors thank Lennart Stenberg for assistance with fieldwork in Sweden, and Anne-Catherine Pasche and Philippe Küpfer for help with laboratory work.

Author information

Authors and Affiliations

  1. Laboratoire de botanique évolutive, Institut de Biologie, Université de Neuchâtel, rue Emile-Argand 11, 2000, Neuchâtel, Switzerland

    Benjamin Dauphin & Jason Grant

  2. Ecology and Evolution Unit, Department of Biology, Université de Fribourg, Chemin du Musée 10, 1700, Fribourg, Switzerland

    Patrik Mráz

  3. Herbarium and Department of Botany, Faculty of Science, Charles University in Prague, Benátská 2, 12801, Prague, Czech Republic

    Patrik Mráz

Authors
  1. Benjamin Dauphin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jason Grant
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Patrik Mráz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Benjamin Dauphin.

Ethics declarations

Funding

B. Dauphin received research Grants from the Fonds Marguerite Wüthrich and A. Matthey-Dupraz at the Université de Neuchâtel, the Fonds Dr. Joachim de Giacomi of the Swiss Academy of Natural Sciences, the Bourse de voyages of the Swiss Academy of Natural Sciences, and the Fonds de donations of the Université de Neuchâtel.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Handling editor: Karol Marhold.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dauphin, B., Grant, J. & Mráz, P. Ploidy level and genome size variation in the homosporous ferns Botrychium s.l. (Ophioglossaceae). Plant Syst Evol 302, 575–584 (2016). https://doi.org/10.1007/s00606-016-1285-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00606-016-1285-7

Keywords

Search

Navigation