Genetic profiling of mistletoe (Viscum album L.) using RAPD-analysis

  • Yu. O. Bilonozhko
  • A. N. Rabokon
  • A. S. Postovoitovа
  • L. O. Kalafat
  • S. M. Privаlikhin
  • N.M. Pirko
  • A. Ye. Demkovych
  • Ya. B. Blume
  • Ya. V. Pirko
DOI: https://doi.org/10.7124/FEEO.v26.1246

Abstract

Aim. The aim of this research was genetic profiling and identification of genetic differences between V. album speciments, growing on deciduous and coniferous species of woody plants using RAPD markers. Methods. The method of polymerase chain reaction (PCR) with random primers (Random Amplification of Polymorphic DNA - RAPD) was used. Amplified DNA fragments were fractionated by electrophoresis in non-denaturing polyacrylamide gel. DNA bands were detected using the staining with silver nitrate. Results. All the studied mistletoe samples were differentiated from each other, and their unique molecular genetic profiles were obtained. 241 amplified DNA fragments were detected in the range from 200 to 2000 bp, 152 fragments (63%) were polymorphic. The samples were divided into two separate groups depending on the type of host plant. Conclusions. The fact that the samples formed two separate clades confirms the assumption that mistletoe, which grow on pine and grow on maple, represents two separate subspecies of V. album.

Keywords: Viscum album L., molecular genetic markers, polymorphism, RAPD.

References

Kolodziejek J., Patykowski J., Kolodziejek R. Distribution, frequency and host patterns of European mistletoe (Viscum album subsp. album) in the major city of Lodz. Poland Biologia. 2013. Vol. 68 (1). P. 55–64. doi: 10.2478/s11756-012-0128-4.

Kim Ch.S., Kim S.Y., Sun B.Y., Yi J.S. A review of the taxonomic and ecological characteristics of Korean mistletoe types (Viscum, Korthalsella, Loranthus and Taxillus). Korean J. Pl. Taxon. 2013. Vol. 43 (2). P. 81–89. doi: 10.11110/kjpt.2013.43.2.81

Ahmed Z., Dutt H.C. Restriction of Viscum album to few phorophytes in a habitat with diverse type of tree species. Austin J Plant Biol. 2015. Vol. 1 (2). P. 101–105.

Schaller G., Urech K., Grazi G., Giannattasio M. Viscotoxin composition of the three European subspecies of Viscum album. Planta Medica. 1998. Vol. 64. P. 677–678. doi: 10.1055/s-2006-957553

Mejnartowicz L. Relationship and genetic diversity of mistletoe (Viscum album L.) subspecies. Acta Soc. Bot. Poloniae. 2006. Vol. 75 (1). P. 39–49. doi: 10.5586/asbp.2006.007

Bishoyi A.K., Sharma A., Kavane A., Geetha K.A. Varietal discrimination and genetic variability analysis of Cymbopogon using RAPD and ISSR markers analysis. Appl. Biochem. Biotechnol. 2016. Vol. 179 (4). P. 659–670. doi: 10.1007/s12010-016-2022-y

Kalkэs O., Okcu M., Okcu Z., Karabulut B. et al. Relationships among some pears genotypes (Pyrus communis L.) based on ISSR and RAPD analysis. Erwerbs-Obstbau. 2016. Vol. 58 (4). P. 259–264. doi: 10.1007/s10341-016-0287-5

Sunar S., Yildirim N., Sengul M., Agar G. Genetic diversity and relationships detected by ISSR and RAPD analysis among Aethionema species growing in Eastern Anatolia (Turkey). C.R. Biol. 2016. Vol. 339. P. 147–151. doi:10.1016/j.crvi.2016.02.006

Green M.R., Sambrook J. Molecular cloning. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press, 2012. 1890 p.

Orlovskaya O.A., Koren L.V., Khotyleva L.V. Evaluation of genetic polymorphism of spring triticale accessions (× Triticosecale Wittmack) based on RAPD and ISSR markers. Vavilov J. Genetics Breeding. 2012. Vol. 16 (1). C. 279–284. [in Russian]

Sambrook J., David W.R. Molecular Сloning: A Laboratory Manual. NY: Cold Spring Harbor, 2001. Vol. 2.

Benbouza H., Jacquemin J-M., Baudoin J.-P., Mergeai G. Optimization of a reliable, fast, cheap and sensitive silver staining method to detect SSR markers in polyacrylamide gel. Biotechnol. Agron. Soc. Environ. 2006. Vol. 10 (2). P. 77–81.

Nei M., Li W.H. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA. 1979. Vol. 76. P. 5269–5273. doi: 10.1073/pnas.76.10.5269

Pavlicek A., Hrda S., Flegr J. FreeTree – Freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap/jackknife analysis of the tree robustness. Application in the RAPD analysis of the genus Frenkelia. Folia Biol. 1999. Vol. 45. P. 97–99.

Hillis D.M., Bull J.J. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biol. 1993. Vol. 42. P. 182–192. doi: 10.1093/sysbio/42.2.182

Rambaut A. FigTree, a graphical viewer of phylogenetic trees. Accessed from: http://tree.bio.ed.ac.uk/software/figtree/