Determination of Sambucus Interspecific Hybrid Structure using Molecular Markers

  • Metka Šiško University of Maribor, Faculty of Agriculture and Life Sciences
  • Anja Ivanuš University of Maribor, Faculty of Agriculture and Life Sciences
  • Anton Ivančič University of Maribor, Faculty of Agriculture and Life Sciences
Keywords: Sambucus, SSR markers, interspecific hybrids, clustering method, molecular markers


Phenotypic and genotypic variations within the genus Sambucus are limited. They could be efficiently increased by genetic recombination involving different species. The aim of the presented investigation was to assess the possibility of using molecular approach (i.e., microsattelites) in determination of unknown hybrid structures. The study involved 47 Sambucus genotypes (parental species and intespecific hybrids), and six microsatellite loci were analysed. The clustering method grouped the analysed genotypes into four main groups. The first main group involved two sub-groups: one with taxons and hybrids involving S. racemosa (in broad sense) and the other involving two botanical varieties of S. nigra. In the second main group, there was a sub-group involving hybrids between S. javanica and S. nigra, and a sub-group involving F1 hybrids between S. javanica and S. ebulus. The third main group contained a sub-group with hybrids between S. javanica and S. nigra ‘Black Beauty’, a sub-group with hybrids involving S. javanica, S. nigra and S. racemosa (miquelii), a sub-group with backcrosses S. javanica × (S. javanica × S. ebulus), and un unknown hybrid. The fourth main group included a subgroup with F1 hybrids S. javanica × S. ebulus, a sub-group involving various taxons of S. racemosa (in broad sense), and a sub-group involving hybrids between S. cerulea and S. javanica, with and without S. nigra. Our study shows that molecular analysis can be helpful in determining some of the unknown but simple interspecific hybrids of Sambucus. In the cases of complex hybrid combinations, the use of SSRs is most probably not the best solution.


Download data is not yet available.

Author Biographies

Metka Šiško, University of Maribor, Faculty of Agriculture and Life Sciences

Hoče, Slovenia. E-mail:

Anja Ivanuš, University of Maribor, Faculty of Agriculture and Life Sciences

Hoče, Slovenia.

Anton Ivančič, University of Maribor, Faculty of Agriculture and Life Sciences

Hoče, Slovenia.


Applequist, 1. W. L. (2015). A brief review of recent controversies in the taxonomy and nomenclature of Sambucus nigra sensu lato. Acta Horticulturae, 1061, 25-33. Retrieved from: https://doi: 10.17660/ActaHortic.2015.1061.1

Atkinson, M. D., & Atkinson, E. (2002). Sambucus nigra L. – Biological flora of the British Isles. No. 225. Journal of Ecology, 90, 895-923.

Böcher, T. W. (1941). Højsommerekskursionen til Brædstrup. Bryrup og Vrads. Botanisk Tidsskrift, 45, 433-439.

Bolli, R. (1994). Dissertationes Botanicae Band 223. Revision of the Genus Sambucus. J. Cramer, Berlin, Stuttgart.

Charlebois, D., Byers, P. L., Finn, C. E., & Thomas, A. L. (2010). Elderberry: botany, horticulture, potential. Horticultural Reviews, 37, 213-280.

Chia, C. L. (1975). A chromosome and thin-layer chromatographic study of the genus Sambucus L. (Doctoral dissertation). Cornell University. College of Agriculture and Life Sciences. Department of Plant Biology. New York. 50 pp.

Clarke, J. B., & Tobutt, K. R. (2006). Development of microsatellite primers and two multiplex polymerase chain reactions for the common elder (Sambucus nigra). Molecular Ecology Notes, 6, 453-455.

Dice, L. R. (1945). Measures of the amount of ecologic association between species. Ecology, 26, 297-302.

Doyle, J. J., & Doyle, J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin,19, 11-15.

Fernald, M. L. (1950). Gray’s manual of botany, 8th ed. New York, NY: American Book Company.

Imenšek, N., Kristl, J., Sem, V., & Ivančič, A. (2020). Elderberry (Sambucus spp.) interspecific hybridization and its impact on fruit oxalates. Plant Breeding, 139(2). Retrieved from:

Koncalová, M. N., Hrib, J., & Jicínská, D. (1983). The embryology of the Sambucus species and hybrids. In Fertilization and Embryogenesis in Ovulated Plants. Proceedings of the VII International Cytoembryological Symposium; Erdelská. O. Ed.; August 1982, Bratislava, Czechoslovakia, 43-47.

Marshall, T. C., Slate, J., Kruuk, L. E. B., & Pemberton, J. M. (1998). Statistical confidence for likelihood-based paternity inference in natural populations. Molecular Ecology Notes, 7, 639-655.

Mikulič-Petkovšek, M., Ivančič, A., Schmitzer, V., Veberič, R., & Štampar, F. (2016). Comparison of major taste compounds and antioxidative properties of fruits and flowers of different Sambucus species and interspecific hybrids. Food Chemistry, 200, 134-140.

Mikulič-Petkovšek, M., Ivančič, A., Todorović, B., Veberič, R., & Štampar, F. (2015a). Fruit phenolic composition of different elderberry species and hybrids. Journal of Food Science, 80, C2180-C2190.

Mikulič-Petkovšek, M., Samoticha, J., Eler, K., Štampar, F., & Veberič, R. (2015b). Traditional elderflower beverages: a rich source of phenolic compounds with high antioxidant activity. Journal of Agricultural and Food Chemistry, 63, 1477-1487.

Mikulič-Petkovšek, M., Schmitzer, V., Slatnar, A., Todorović, B., Veberič, R., Štampar, F., & Ivančič, A. (2014). Investigation of anthocyanin profile of four elderberry species and interspecific hybrids. Journal of Agricultural and Food Chemistry, 62(24), 5573-5580.

Nilsson, A. (1987). Hybriden mellan fläder och druvfläder funnen i Skåne. Svensk Botanisk Tidskrift, 81, 174-175.

Perrier, X., & Jacquemoud-Collet, J. P. (2006). DARwin software. Retrieved from:

Saitou, N., & Nei, M. (1987). The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology Evolution, 4, 406-425.

Shokrzadeh, M., & Saravi, S. S. S. (2010). The chemistry, pharmacology and clinical properties of Sambucus ebulus: a review. Journal of Medicinal Plants Research, 4, 95-103.

Simonovik, B. (2007). Genetic evaluation of interspecific hybrids in the genus Sambucus and establishment of selected plant tissue culture techniques in common elder (S. nigra L.) (Doctoral dissertation). Biotechnical Faculty, University of Ljubljana, Ljubljana.

Simonovik, B., Ivančič, A., Jakše, J., & Bohanec, B. (2007). Production and genetic evaluation of interspecific hybrids within the genus Sambucus. Plant Breeding, 126(6), 628-633.

Tessier, C., David, J., This, P., Boursiquot, J. M., & Charrier, A. (1999). Optimization of the choice of molecular markers for varietal identification in Vitis vinifera L. Theoretical and Applied Genetics, 98, 171-177.

Todorović, B., Mikulič-Petkovšek, M., Štampar, F., & Ivančič, A. (2017). Phenolic compounds in floral infusions of various Sambucus species and their interspecific hybrids. Turkish Journal of Agriculture and Forestry, 41(2), 154-164.

Vlachojannis, J. E., Cameron, M., & Chrubasik, S. (2010). A systematic review on the sambuci fructus effect and efficacy profiles. Phytotherapy Research, 24(1), 1-8.

von Linné, C. (Caroli Linnaei). 1753. Species plantarum: exhibentes plantas rite cognitas, ad genera relatas, cum differentiis specificis, nominibus trivialibus, synonymis selectis, locis natalibus, secundum systema sexuale digestas... Holmiae; Impensis Laurentii Salvii. Retrieved from:

von Schwerin, F. G. (1920). Revisio generis Sambucus. Mitteilungen der Deutschen dendrologischen gesellschaft, 29, 194-231.

Wenga, J. -R., Linb, C. -S., Laic, H. -C., Line, Y. -P., Wange, C. -Y., Tsaie, Y.-C., Wuf, K. -C., Huangg, S. -H., & Line, C. - W. (2019). Antiviral activity of Sambucus Formosana Nakai ethanol extract and related phenolic acid constituents against human coronavirus NL63. Virus Research, 273, 1-8.

Weston, R. (1775). The English flora: or, a catalogue of trees, shrubs, plants and fruits, natives as well as exotics, cultivated, for use or ornament, in the English nurseries, greenhouses and stoves: arranged according to the Linnaean system: with the Latin trivial, and common English names, and an English index referring to the Latin names. Also a general catalogue of seeds for the kitchen-garden, flower-garden, grass-lands etc., usually raised for sale, and those annually imported from America. Printed for the Author and sold by: Millan, J., Robson and Co. New Bond-Street, Carnan, T., St. Paul’s Church-Yard, Dilly, E., and C., London.

Winge, Ö. (1944). The Sambucus hybrid S. nigra × S. racemosa. Comptesrendus des travaux du laboratories Carlsberg. Sér Physiolog, 24, 73-78.

Zulini, L., Russo, M., & Peterlunger, E. (2002). Genotyping wine and table grape cultivars from Apulia (Southern Italy) using microsatellite markers. Vitis, 41, 183-187.

How to Cite
Šiško M., Ivanuš A., & Ivančič A. (2020). Determination of Sambucus Interspecific Hybrid Structure using Molecular Markers. Agricultura Scientia, 16(1-2), 1-10.