Planta Med 2003; 69(10): 939-944
DOI: 10.1055/s-2003-45104
Original Paper
Analytical Methods
© Georg Thieme Verlag Stuttgart · New York

Molecular Genetic Survey of European Mistletoe (Viscum album) Subspecies with Allele-Specific and dCAPS Type Markers Specific for Chloroplast and Nuclear DNA Sequences

Arkadiusz Piotrowski1 , J. Renata Ochocka1 , Justyna Stefanowicz1 , Maria Łuczkiewicz2
  • 1Department of Biology and Pharmaceutical Botany, Medical University of Gdańsk, Gdańsk, Poland
  • 2Department of Pharmacognosy, Medical University of Gdańsk, Gdańsk, Poland
Further Information

Publication History

Received: March 4, 2003

Accepted: May 3, 2003

Publication Date:
02 December 2003 (online)

Abstract

The qualitative and quantitative content of mistletoe metabolites, and bioactivity of extracts is related to the subspecies of Viscum album L. These were indicated to be genetically distinct and host specific. We aimed to check (i) whether the specificity is strict and (ii) how frequently hybridization occurs among the subspecies. We designed two sets of allele-specific and dCAPS molecular genetic markers that would facilitate identification of Viscum album L. subspecies and their hybrid derivatives on the basis of chloroplast trnH(GUG)-trnK(UUU) and nuclear rDNA ITS1&2 sequences. Out of 118 plants surveyed, 103 displayed characteristics that confirmed strict host specificity of the subspecies, in addition, the results were compliant between nuclear and chloroplast markers showing no indication of hybridization among subspecies. From 15 samples that showed deviations from this model 13 came from the Mediterranean Sea basin, and only two originated from Central and Western Europe.

Abbreviations

dCAPS:derived Cleaved Amplified Polymorphic Sequence

ITS1&2:Internal Transcribed Spacers 1&2

MAMA:Mismatch Amplification Mutation Assay

References

  • 1 Büssing A, Schietzel M. Apoptosis-inducing properties of Viscum album L. extracts from different host trees, correlate with their content of toxic mistletoe lectins.  Anticancer Res. 1999;  19 23-8
  • 2 Ribereau-Gayon G, Dumont S, Muller C, Jung M L, Poindron P, Anton R. Mistletoe lectins I, II and III induce the production of cytokines by cultured human monocytes.  Cancer Lett. 1996;  109 33-8
  • 3 Ochocka J R, Piotrowski A. Biologically active compounds from European mistletoe (Viscum album L.)  Can J Plant Pathol. 2002;  24 21-8
  • 4 Schaller G, Urech K, Grazi G, Giannattasio M. Viscotoxin composition of the three European subspecies of Viscum album .  Planta Med.. 1998;  64 677-8
  • 5 Khwaja T A, Dias C B, Pentecost S. Recent studies on the anticancer activities of mistletoe (Viscum album) and its alkaloids.  Oncology. 1986;  43 Suppl 1 42-50
  • 6 Stein G M, Schietzel M, Bussing A. Mistletoe in immunology and the clinic (short review).  Anticancer Res. 1998;  18 3247-9
  • 7 Zuber D, Widmer A. Genetic evidence for host specificity in the hemi-parasitic Viscum album L. (Viscaceae).  Mol Ecol. 2000;  9 1069-73
  • 8 Doyle J J, Doyle J L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue.  Phytochem Bull. 1987;  19 11-5
  • 9 Yang M, Zhang D, Liu J, Zheng J. A molecular marker that is specific to medicinal rhubarb based on chloroplast trnL/trnF sequences.  Planta Med. 2001;  67 784-6
  • 10 Cha R S, Zarbl H, Keohavong P, Thilly W G. Mismatch amplification mutation assay (MAMA): application to the c-H-ras gene.  PCR Methods Appl. 1992;  2 14-20
  • 11 Michaels S D, Amasino R M. A robust method for detecting single-nucleotide changes as polymorphic markers by PCR.  Plant J. 1998;  14 381-5
  • 12 Neff M M, Neff J D, Chory J, Pepper A E. dCAPS, a simple technique for the genetic analysis of single nucleotide polymorphisms: experimental applications in Arabidopsis thaliana genetics.  Plant J. 1998;  14 387-92
  • 13 Thompson J D, Higgins D G, Gibson T J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.  Nucleic Acids Res. 1994;  22 4673-80
  • 14 Hall T A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT.  Nucl Acids Symp Ser. 1999;  41 95-8
  • 15 Rice P, Longden I, Bleasby A. EMBOSS: the European Molecular Biology Open Software Suite.  Trends Genet. 2000;  16 276-7
  • 16 Soltis D E, Soltis P S. Choosing an approach and an appropriate gene for phylogenetic analysis. In: DE Soltis, PS Soltis, JJ Doyle (editors) Molecular Systematics of Plants II. DNA Sequencing. Massachusetts, USA; Kluwer Academic Publishers 2000: pp. 1-42
  • 17 Nickrent D L, Duff R J, Colwell A E, Wolfe A D, Young N D, Steiner K E, dePamphilis C W. Molecular phylogenetic and evolutionary studies of parasitic plants. In: DE Soltis, PS Soltis, JJ Doyle (editors) Molecular Systematics of Plants II. DNA Sequencing. Massachusetts, USA; Kluwer Academic Publishers 2000: pp. 211-41
  • 18 Stypinski P. Biology and ecology of the European mistletoe (Viscum album, Viscaceae) in Poland.  Fragm Flor Geobot Ser Polonica. 1997;  (Supplement 1) 3-115
  • 19 Jerome C A, Ford B A. The discovery of three genetic races of the dwarf mistletoe Arceuthobium americanum (Viscaceae) provides insight into the evolution of parasitic angiosperms.  Mol Ecol. 2002;  11 387-405
  • 20 Martin-Cordero C, Lopez-Lazaro M, Agudo M A, Navarro E, Trujillo J, Ayuso M J. A cytotoxic diarylheptanoid from Viscum cruciatum .  Phytochemistry. 2001;  58 567-9

Arkadiusz Piotrowski

Department of Biology and Pharmaceutical Botany

Medical University of Gdansk

ul. Hallera 107

80-416 Gdansk

Poland

Phone: +48-58-3493214

Fax: +48-58-3493211

Email: arpiotr@amg.gda.pl