Plant Systematics and Evolution

, Volume 281, Issue 1–4, pp 77–86 | Cite as

Identifying a mysterious aquatic fern gametophyte

  • Fay-Wei LiEmail author
  • Benito C. Tan
  • Volker Buchbender
  • Robbin C. Moran
  • Germinal Rouhan
  • Chun-Neng WangEmail author
  • Dietmar QuandtEmail author
Original Article


Süßwassertang, a popular aquatic plant that is sold worldwide in aquarium markets, has been long considered a liverwort because of its ribbon-like thallus. However, its antheridia are remarkably fern-like in morphology. To corroborate the hypothesis that Süßwassertang is a fern gametophyte and to determine its closest relative, we have sequenced five chloroplast regions (rbcL, accD, rps4trnS, trnL intron, and trnL-F intergenic spacer), applying a DNA-based identification approach. The BLAST results on all regions revealed that Süßwassertang is a polypod fern (order: Polypodiales) with strong affinities to the Lomariopsidaceae. Our phylogenetic analyses further showed that Süßwassertang is nested within the hemi-epiphytic fern genus Lomariopsis (Lomariopsidaceae) and aligned very close to L. lineata. Our study brings new insights on the unexpected biology of Lomariopsis gametophytes—the capacity of retaining a prolonged gametophytic stage under water. It is of great interest to discover that a fern usually known to grow on trees also has gametophytes that thrive in water.


Aquarium DNA barcoding DNA-based identification Gametophyte Fern Lomariopsis Lomariopsidaceae 



The authors thank Li-Yaung Kuo for laboratory help, Tien-Chuan Hsu for collecting Taiwanese Lomariopsis spectabilis, and Dr. Wen-Liang Chiou (Taiwan Forestry Research Institute) for valuable comments.


  1. Atkinson LR (1973) The gametophyte and family relationships. Bot J Linn Soc 67[Suppl 1]:73–90Google Scholar
  2. Chase MW, Salamin N, Wilkinson M, Dunwell JM, Kesanakurthi RP, Haidar N, Savolainen V (2005) Land plants and DNA barcodes:short-term and long-term goals. Philos Trans R Soc Lond B Biol Sci 360:1889–1895PubMedCrossRefGoogle Scholar
  3. Chase MW, Cowan RS, Hollingsworth PM, van den Berg C, Madriñán S, Petersen G, Seberg O, Jørgsensen T, Cameron KM, Carine M, Pedersen N, Hedderson TAJ, Conrad F, Salazar GA, Richardson JE, Hollingsworth ML, Barraclough TG, Kelly L, Wilkinson M (2007) A proposal for a standardised protocol to barcode all land plants. Taxon 56:295–299Google Scholar
  4. Cox CJ, Goffinet B, Newton AE, Shaw AJ, Hedderson TAJ (2000) Phylogenetic relationships among the diplolepideous-alternate mosses (Bryidae) inferred from nuclear and chloroplast DNA sequences. Bryologist 103:224–241CrossRefGoogle Scholar
  5. Dassler CL, Farrar DR (1997) Significance of form in fern gametophytes: clonal, gemmiferous gametophytes of Callistopteris baueriana (Hymenophyllaceae). Int J Plant Sci 158:622–639CrossRefGoogle Scholar
  6. Farrar DR (1967) Gametophytes of four tropical fern genera reproducing independently of their sporophytes in the southern Appalachians. Science 155:1266–1267PubMedCrossRefGoogle Scholar
  7. Farrar DR (1990) Species and evolution in asexually reproducing independent fern gametophytes. Syst Bot 15:98–111CrossRefGoogle Scholar
  8. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  9. Ford CS, Ayres KL, Toomey N, Haider N, Stahl JV, Kelly LJ, Wikström N, Hollingsworth PM, Duff RJ, Hoot SB, Cowan RS, Chase MW, Wilkinson MJ (2009) Selection of candidate coding DNA barcoding regions for use on land plants. Bot J Linn Soc 159:1–11CrossRefGoogle Scholar
  10. Hollingsworth ML, Clark AA, Forrest LL, Richardson J, Pennington RT, Long DG, Cowan R, Chase MW, Gaudeul M, Hollingsworth PM (2009) Selecting barcoding loci for plants: evaluation of seven candidate loci with species-level sampling in three divergent groups of land plants. Mol Ecol Resour 9:439–457Google Scholar
  11. Huelsenbeck JP, Ronquist F, Nielsen R, Bollback JP (2001) Bayesian inference of phylogeny and its impact on evolutionary biology. Science 294:2310–2314PubMedCrossRefGoogle Scholar
  12. Huelsenbeck JP, Larget B, Miller RE, Ronquist F (2002) Potential applications and pitfalls of Bayesian inference of phylogeny. Syst Biol 51:673–688PubMedCrossRefGoogle Scholar
  13. Johnson GN, Rumsey FJ, Headley AD, Sheffield E (2000) Adaptions to extreme low light in the fern Trichomanes speciosum. New Phytol 148:423–431CrossRefGoogle Scholar
  14. Kelchner SA (2000) The evolution of non-coding chloroplast DNA and its application in plant systematics. Ann Mo Bot Gard 87:482–498CrossRefGoogle Scholar
  15. Kress WJ, Erickson DL (2007) A two-locus global DNA barcode for land plants: the coding rbcL gene complements the non-coding trnH-psbA spacer region. PLoS ONE 2:e508PubMedCrossRefGoogle Scholar
  16. Kress WJ, Wurdack KJ, Zimmer EA, Weigt LA, Janzen DH (2005) Use of DNA barcodes to identify flowering plants. Proc Natl Acad Sci USA 102:8369–8374PubMedCrossRefGoogle Scholar
  17. Lahaye R, Bank M, Borgarin D, Warner J, Pupulin F, Gigot G, Maurin O, Duthoit S, Barraclough TG, Savolainen V (2008) DNA barcoding the floras of biodiversity hotspots. Proc Natl Acad Sci USA 105:2923–2928PubMedCrossRefGoogle Scholar
  18. Lewis LA, Mishler BD, Vilgalys R (1997) Phylogenetic relationships of the liverworts (Hepaticae), a basal embryophyte lineage, inferred from nucleotide sequence data of the chloroplast gene rbcL. Mol Phylogenet Evol 7:377–393PubMedCrossRefGoogle Scholar
  19. Li CX, Lu SG (2006) Phylogenetic analysis of Dryopteridaceae based on chloroplast rbcL sequences. Acta Phytotax Sin 44:503–515CrossRefGoogle Scholar
  20. Müller K (2004) PRAP—calculation of Bremer support for large data sets. Mol Phylogenet Evol 31:780–782PubMedCrossRefGoogle Scholar
  21. Müller J, Müller K (2004) TreeGraph: automated drawing of complex tree figures using an extensible tree description format. Mol Ecol Notes 4:6–788CrossRefGoogle Scholar
  22. Müller K, Quandt D, Müller J, Neinhuis C (2005) PhyDE® 0995. Phylogenetic Data Editor. Available at:
  23. Nadot SN, Bajon R, Lejeune B (1994) The chloroplast gene rps4 as a tool for the study of Poaceae phylogeny. Plant Syst Evol 191:27–38CrossRefGoogle Scholar
  24. Nayar BK, Kaur S (1971) Gametophytes of homosporous ferns. Bot Rev 37:295–396CrossRefGoogle Scholar
  25. Nitta J (2008) Exploring the utility of three plastid loci for biocoding the filmy ferns (Hymenophyllaceae) of Morea. Taxon 57:725–736Google Scholar
  26. Nixon KC (1999) The parsimony ratchet, a new method for rapid parsimony analysis. Cladistics 15:407–411CrossRefGoogle Scholar
  27. Quandt D, Stech M (2003) Molecular systematics of bryophytes in context of land plant evolution. In: Sharma AK, Sharma A (eds) Plant genome: biodiversity and evolution, vol 1. Science Publ, Enfield, pp 267–295Google Scholar
  28. Quandt D, Stech M (2004) Molecular evolution and phylogenetic utility of the chloroplast trnT-trnF region in bryophytes. Plant Biol 6:545–554PubMedCrossRefGoogle Scholar
  29. Quandt D, Müller K, Huttunen S (2003) Characterisation of the chloroplast DNA psbT-H region and the influence of dyad symmetrical elements on phylogenetic reconstructions. Plant Biol 5:400–410CrossRefGoogle Scholar
  30. Quandt D, Müller K, Stech M, Hilu KW, Frey W, Frahm JP, Borsch T (2004) Molecular evolution of the chloroplast trnL-F region in land plants. Monogr Syst Bot Mo Bot Gard 98:13–37Google Scholar
  31. Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574PubMedCrossRefGoogle Scholar
  32. Rouhan G, Hanks JG, McClelland D, Moran RC (2007) Preliminary phylogenetic analysis of the fern genus Lomariopsis (Lomariopsidaceae). Brittonia 59:115–128CrossRefGoogle Scholar
  33. Rumsey FJ, Vogel JC, Russell SJ, Barrett JA, Gibby M (1999) Population structure and conservation biology of the endangered fern Trichomanes speciosum Willd (Hymenophyllaceae) at its northern distributional limit. Biol J Linn Soc 66:333–344Google Scholar
  34. Sato T (1982) Phenology and wintering capacity of sporophytes and gametophytes of ferns native to northern Japan. Oecologia 55:53–61CrossRefGoogle Scholar
  35. Schneider H, Schuettpelz E (2006) Identifying fern gametophytes using DNA sequences. Mol Ecol Notes 6:989–991CrossRefGoogle Scholar
  36. Schneider H, Schuettpelz E, Pryer KM, Cranfill R, Magallón Lupia R (2004) Ferns diversified in the shadow of angiosperms. Nature 428:553–557PubMedCrossRefGoogle Scholar
  37. Skog JE, Mickel JT, Moran RC, Volovsek M, Zimmer EA (2004) Molecular studies of representative species in the fern genus Elaphoglossum (Dryopteridaceae) based on cpDNA sequences rbcL, trnL-F, and rps4-trnS. Int J Plant Sci 165:1063–1075CrossRefGoogle Scholar
  38. Smith AR, Cranfill RB (2002) Infrafamilial relationships of the thelypteroid ferns (Thelypteridaceae). Am Fern J 92:131–149CrossRefGoogle Scholar
  39. Souza-Chies TT, Bittar G, Nadot S, Carter L, Besin E, Lejeune B (1997) Phylogenetic analysis of Iridaceae with parsimony and distance methods using the plastid gene rps4. Plant Syst Evol 204:109–123CrossRefGoogle Scholar
  40. Swofford DL (2002) PAUP*: phylogenetic analysis using parsimony (*and other methods) Version 40b10. Sinauer, SunderlandGoogle Scholar
  41. Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three noncoding regions of chloroplast DNA. Plant Mol Biol 17:1105–1109PubMedCrossRefGoogle Scholar
  42. Taberlet P, Coissac E, Pompanon F, Gielly L, Miquel C, Valentini A, Vermat T, Corthier G, Brochmann C, Willerslev E (2006) Power and limitations of the chloroplast trnL (UAA) intron for plant DNA barcoding. Nucleic Acids Res 35:e14PubMedCrossRefGoogle Scholar
  43. Tsutsumi C, Kato M (2005) Molecular phylogenetic study on Davalliaceae. Fern Gaz 17:147–162Google Scholar
  44. Tsutsumi C, Kato M (2006) Evolution of epiphytes in Davalliaceae and related ferns. Bot J Linn Soc 151:495–510CrossRefGoogle Scholar
  45. Wakasugi T, Tsudzuki J, Ito S, Nakashima K, Tsudzuki T, Sugiura M (1994) Loss of all ndh genes as determined by sequencing the entire chloroplast genome of the black pine Pinus thunbergii. Proc Natl Acad Sci USA 91:9794–9798PubMedCrossRefGoogle Scholar
  46. Wang C-N, Moller M, Cronk QCB (2004) Phylogenetic position of Titanotrichum oldhamii (Gesneriaceae) inferred from four different gene regions. Syst Bot 29:407–418CrossRefGoogle Scholar
  47. Watkins JE Jr, Mack MC, Sinclair TR, Mulkey SS (2007) Ecological and evolutionary consequences of desiccation tolerance in tropical fern gametophytes. New Phytol 176:708–717PubMedCrossRefGoogle Scholar
  48. Wolf PG, Rowe CA, Sinclair RB, Hasebe M (2003) Complete nucleotide sequence of the chloroplast genome from a leptosporangiate fern, Adiantum capillus-veneris L. DNA Res 10:59–65PubMedCrossRefGoogle Scholar
  49. Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Life ScienceNational Taiwan UniversityTaipeiTaiwan
  2. 2.The HerbariumSingapore Botanic GardensSingaporeSingapore
  3. 3.Plant Phylogenetics and Phylogenomics Group, Institute of BotanyDresden University of TechnologyDresdenGermany
  4. 4.The New York Botanical GardenBronx, New YorkUSA
  5. 5.Muséum National d’Histoire Naturelle, UMR 7205Herbier NationalParisFrance
  6. 6.Institute of Ecology and Evolutionary BiologyNational Taiwan UniversityTaipeiTaiwan
  7. 7.Nees Institute for Biodiversity of PlantsUniversity of BonnBonnGermany

Personalised recommendations