ORIGINAL PAPER Whole genome amplification and microsatellite genotyping of herbarium DNA revealed the identity of an ancient grapevine cultivar Nenad Malenica & Silvio Šimon & Višnja Besendorfer & Edi Maletić & Jasminka Karoglan Kontić & Ivan Pejić Received: 2 March 2011 /Revised: 30 June 2011 /Accepted: 2 July 2011 /Published online: 22 July 2011 # Springer-Verlag 2011 Abstract Reconstruction of the grapevine cultivation his- tory has advanced tremendously during the last decade. Identification of grapevine cultivars by using microsatellite DNA markers has mostly become a routine. The parentage of several renowned grapevine cultivars, like Cabernet Sauvignon and Chardonnay, has been elucidated. However, the assembly of a complete grapevine genealogy is not yet possible because missing links might no longer be in cultivation or are even extinct. This problem could be overcome by analyzing ancient DNA from grapevine herbarium specimens and other historical remnants of once cultivated varieties. Here, we present the first successful genotyping of a grapevine herbarium speci- men and the identification of the corresponding grape- vine cultivar. Using a set of nine grapevine microsatellite markers, in combination with a whole genome amplifi- cation procedure, we found the 90-year-old Tribidrag herbarium specimen to display the same microsatellite profile as the popular American cultivar Zinfandel. This work, together with information from several historical documents, provides a new clue of Zinfandel cultivation in Croatia as early as the beginning of fifteenth century, under the native name Tribidrag. Moreover, it emphasizes substantial information potential of existing grapevine and other herbarium collections worldwide. Keywords SSR . aDNA . Vitis vinifera . Tribidrag . Zinfandel . Herbarium Introduction Worldwide, thousands of grapevine herbarium collections are preserving both ampelographic and nomenclatural information about old grapevine cultivars. Many of the specimens are probably the last remnants of forgotten or extinct cultivars, which are no longer grown. If these collections could be successfully genotyped, they would have a huge potential for discovering the missing links in genealogy of grapevine cultivars and history of grapevine cultivation. In addition to herbaria, various grapevine archaeological remnants, such as woody parts or seeds, could serve a similar purpose (This et al. 2006). Since both growers and consumers prefer old traditional grapevine varieties, the above raised issues have not only scientific but also a potentially important marketing dimension. The field of ancient DNA (aDNA) research is devoted to gaining valuable information from old biological speci- mens. Ancient DNA research is steadily developing for the last 25 years. Despite occasional false-positive results being published in high-profile journals (Cooper and Poinar 2000), the field came to the current state of art where entire ancient genomes can reliably be sequenced, as Communicated by: Sven Thatje N. Malenica (*) : V . Besendorfer Faculty of Science, Department of Molecular Biology, University of Zagreb, Horvatovac 102a, 10000 Zagreb, Croatia e-mail: malenica@zg.biol.pmf.hr S. Šimon : I. Pejić Faculty of Agriculture, Department of Plant Breeding, Genetics and Biometrics, University of Zagreb, Svetošimunska 25, 10000 Zagreb, Croatia E. Maletić : J. Karoglan Kontić Faculty of Agriculture, Department of Viticulture and Enology, University of Zagreb, Svetošimunska 25, 10000 Zagreb, Croatia Naturwissenschaften (2011) 98:763–772 DOI 10.1007/s00114-011-0826-8