Skip to main content
SpringerLink
Log in

Genetic variability and relationships among interspecific hybrid cultivars and parental species of Paphiopedilum via ribosomal DNA sequence analysis

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

Abstract

The aim of the present study was to investigate genetic variability and relationships among several hybrid cultivars and parental species of Paphiopedilum for the purposes of further breeding. Genetic information was obtained by means of phylogenetic analysis, using nuclear ribosomal DNA (rDNA) nucleotide sequence data. Nuclear rDNA was derived from internal transcribed spacer (ITS) regions, and the 5.8S regions were amplified by polymerase chain reaction. The resulting nucleotide sequences were examined and compared for genetic variability, and the relationships were analyzed by pair-wise multiple alignment. ITS region analysis revealed the following lengths of the ITS1, ITS2, and 5.8S regions: 382–428, 207–215, and 170 bp, respectively. Sequence identity at the ITS site, including the 5.8S region, ranged from 75.9 % between P. emersonii and P. sanderianum, to 99.4 % between Paphiopedilum ‘Hamana Wave’ and Paphiopedilum ‘Kolosand’. Next, we investigated whether each of the analyzed hybrid cultivars could be identified and differentiated at the interspecies or intraspecies levels. Our phylogenetic tree revealed that hybrids derived from the subgenus Brachypetalum were clearly differentiated from those derived from the subgenus Paphiopedilum. Moreover, interspecific hybrids showed unique phylogenetic positioning among parental species, according to the ITS sequence.

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

  • Aceto S, Caputo P, Cozzolino S, Gaudio L (1999) Phylogeny and evolution of Orchis and allied genera based on ITS DNA variation: morphological gaps and molecular continuity. Mol Phylogenet Evol 13:67–76

    Article  PubMed  CAS  Google Scholar 

  • Altschul SF, Madden TL, Schaffer A, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed  CAS  Google Scholar 

  • Baldwin BG (1992) Phylogenetic utility of the internal transcribed spacers of nuclear ribosomal DNA in plants: an example from the compositae. Mol Phylogenet Evol 1:3–6

    Article  PubMed  CAS  Google Scholar 

  • Baldwin BG (1993) Molecular phylogenetics of Calycadenia (compositae) based on ITS sequences of nuclear ribosomal DNA: chromosomal and morphological evolution reexamined. Am J Bot 80:222–238

    Article  CAS  Google Scholar 

  • Baldwin BG, Sanderson MJ, Porter JM, Wojciechowski MF, Campbell CS, Donoghue MJ (1995) The ITS region of nuclear ribosomal DNA: a valuable source of evidence on angiosperm phylogeny. Ann Mo Bot Gard 82:247–277

    Article  Google Scholar 

  • Charmet G, Ravel C, Balfourier F (1997) Phylogenetic analysis in the Festuca–Lolium complex using molecular markers and ITS rDNA. Theor Appl Genet 94:1038–1046

    Article  CAS  Google Scholar 

  • Chase MW (2005) Classification of Orchidaceae in the age of DNA data. Curtis’s Bot Mag 22:2–7

    Article  Google Scholar 

  • Chen SC, Tsi ZH (1984) On Paphiopedilum malipoense sp. nov—an intermediate form between Paphiopedilum and Cypripedium. Acta Phytotax Sin 22:119–124

    CAS  Google Scholar 

  • Chung SY, Choi SH, Kim MJ, Yoon KE, Lee GP, Lee JS, Ryu KH (2006) Genetic relationship and differentiation of Paphiopedilum and Phragmipedium based on RAPD analysis. Sci Hort 103:153–159

    Article  Google Scholar 

  • Cox AV, Bennett ST, Parokonny AS, Kenton A, Callimassia MA, Bennett MD (1993) Comparison of plant telomere locations using a PCR-generated synthetic probe. Ann Bot 72:239–247

    Article  CAS  Google Scholar 

  • Cox AV, Pridgen AM, Albert VA, Chase MW (1997) Phylogenetics of the slipper orchids (Cypripedioideae: Orchidaceae): nuclear rDNA ITS sequences. Plant Syst Evol 208:187–223

    Article  Google Scholar 

  • Douzery EJP, Pridgeon AM, Kores P, Linder HP, Kurzweil H, Chase MW (1999) Molecular phylogenetics of Diseae (Orchidaceae): a contribution from nuclear ribosomal ITS sequences. Am J Bot 86:887–899

    Article  PubMed  CAS  Google Scholar 

  • Downie SR, Deborah S, Katz-Downie S (1996) A molecular phylogeny of Apiaceae subfamily Apioideae: evidence from nuclear ribosomal DNA internal transcribed spacer sequences. Am J Bot 83:234–251

    Article  CAS  Google Scholar 

  • Goh MWK, Kumar PP, Lim SH, Tan HTW (2005) Random amplified polymorphic DNA analysis of the moth orchids, Phalaenopsis (Epidendroideae: Orchidaceae). Euphytica 141:11–22

    Article  CAS  Google Scholar 

  • Intrieri MC, Muleo R, Buiatti M (2006) Chloroplast DNA polymorphisms as molecular markers to identify cultivars of Olea europaea L. J Hort Sci Biotech 82:109–113

    Google Scholar 

  • Jobst J, King K, Hemleben V (1998) Molecular evolution of the internal transcribed spacers (ITS1 and ITS2) and phylogenetic relationships among species of the family Cucurbitaceae. Mol Phylogenet Evol 9:204–219

    Article  PubMed  CAS  Google Scholar 

  • Karasawa K (1979) Karyomorphological studies in Paphiopedilum, Orchidaceae. Bull Hiroshima Bot Gard 2:1–149

    Google Scholar 

  • Koopowitz H (2000) A revised checklist of the genus Paphiopedilum. Orchid Digest Oct.-Nov.-Dec. 155–179

  • Lee YI, Chang FC, Chung MC (2011) Chromosome pairing affinities in interspecific hybrids reflect phylogenetic distances among lady’s slipper orchids (Paphiopedilum). Ann Bot 108:113–121

    Article  PubMed  Google Scholar 

  • Marshall JA, Knapp S, Davey MR, Power JB, Cocking EC, Bennet MD, Cox AV (2001) Molecular systematic of Solanum section Lycopersicum (Lycopersicon) using the nuclear ITS rDNA region. Theor Appl Genet 103:1216–1222

    Article  CAS  Google Scholar 

  • Nielsen LR, Siegismund HR (1999) Interspecific differentiation and hybridization in Vanilla species (Orchidaceae). Heredity 83:560–567

    Article  PubMed  Google Scholar 

  • Palacios C, Rossello JA, Gonzalez-Candelas F (2000) Study of the evolutionary relationships among Limonium species (Plumbaginaceae) using nuclear and cytoplasmic molecular markers. Mol Phylogenet Evol 94:232–249

    Article  Google Scholar 

  • Sang T, Crawford DJ, Stuessy TF (1995) Documentation of reticulate evolution in peonies (Paeonia) using internal transcribed spacer sequences of nuclear ribosomal DNA: implication for biogeography and concerted evolution. Proc Natl Acad Sci USA 92:6813–6817

    Article  PubMed  CAS  Google Scholar 

  • Sanger F, Niklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A 74:5463–5467

    Article  PubMed  CAS  Google Scholar 

  • Schlegel M, Steinbruck G, Hahn K, Rottger B (1989) Interspecific relationship of ten European orchid species as revealed by enzyme electrophoresis. Plant Syst Evol 163:107–119

    Article  Google Scholar 

  • Soliva M, Kocyan A, Widmer A (2001) Molecular phylogenetics of the sexually deceptive orchid genus Ophrys (Orchidaceae) based on nuclear and chloroplast DNA sequences. Mol Phylogenet Evol 20:78–88

    Article  PubMed  CAS  Google Scholar 

  • Soltis PS, Soltis DE, Wolf PG, Nickrent DL, Chaw SM, Chapman RL (1999) The phylogeny of land plants inferred from 18S rDNA sequences: pushing the limits of rDNA signal? Mol Biol Evol 16:1774–1784

    Article  PubMed  CAS  Google Scholar 

  • Suh Y, Thien L, Reeve HF, Zimmer EA (1993) Molecular evolution and phylogenetic implications of internal transcribed spacer sequences of ribosomal DNA in Winteraceae. Am J Bot 80:1042–1055

    Article  CAS  Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson T (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  • Tsai CC, Peng CI, Huang SC, Huang PL, Chou CH (2004) Determination of the genetic relationship of Dendrobium species (Orchidaceae) in Taiwan based on the sequence of the internal transcribed spacer of ribosomal DNA. Sci Hort 101:315–325

    Article  CAS  Google Scholar 

  • Umehara M, Sueyoshi T, Shimomura K, Iwai M, Shigyo M, Hirashima K, Nakahara T (2006) Interspecific hybrids between Allium fistulosum and Allium schoenoprasum reveal carotene-rich phenotype. Euphytica 148:295–301

    Article  CAS  Google Scholar 

  • Wang QD, Zhang T, Wang JB (2007) Phylogenetic relationships and hybrid origin of Potamogeton species (Potamogetonaceae) distributed in China: insights from the nuclear ribosomal internal transcribed spacer sequence (ITS). Pl Syst Evol 267:65–78

    Article  CAS  Google Scholar 

  • Whitten WM, Williams NH, Chase MW (2000) Subtribal and generic relationships of Maxillarieae (Orchidaceae) with emphasis on Stanhopeinae: combined molecular evidence. Am J Bot 87:1842–1856

    Article  PubMed  CAS  Google Scholar 

  • Wojciechowski MF, Sanderson MJ, Baldwin BG, Donoghue MJ (1993) Monophyly of aneuploid Astragalus (Fabaceae): evidence from nuclear ribosomal DNA internal transcribed spacer sequences. Am J Bot 80:711–722

    Article  CAS  Google Scholar 

  • Yang YW, Lai KN, Tai PY, Ma DP, Li WH (1999) Molecular phylogenetic studies of Brassica, Rorippa, Arabidopsis and allied genera based on the internal transcribed spacer region of 18S–25S rDNA. Mol Phylogenet Evol 13:455–462

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a research grant from Seoul Women’s University (2011).

Author information

Authors and Affiliations

  1. Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women’s University, Seoul, 139-774, Korea

    So Young Chung & Sun Hee Choi

Authors
  1. So Young Chung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sun Hee Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sun Hee Choi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chung, S.Y., Choi, S.H. Genetic variability and relationships among interspecific hybrid cultivars and parental species of Paphiopedilum via ribosomal DNA sequence analysis. Plant Syst Evol 298, 1897–1907 (2012). https://doi.org/10.1007/s00606-012-0689-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00606-012-0689-2

Keywords

Search

Navigation