Genetic identification of 200-year-old Serbian grapevine herbarium
Botanist Andreas Raphael Wolny collected a grapevine herbarium from 1812-1824 in Sremski Karlovci (wine region of Vojvodina, Serbia), which represents local cultivated grapevine diversity before the introduction of grape phylloxera in the region. The herbarium comprises over 100 samples organized into two subcollections based on berry colour (red and white varieties), totaling 47 different grape varieties. The objective of this study was to investigate the historical varietal assortment of Balkan and Pannonian winegrowing areas with long viticulture traditions. Obtaining DNA from more than 100 years old plant remains requires the use of ancient (or archive) DNA (aDNA) extraction procedures, adapted to aDNA conditions (typically degraded, damaged, and contaminated with environmental DNA). Here, we used a protocol for recovering ultrashort DNA molecules from herbarized leaves in facilities tailored for aDNA extraction. In 84 of 103 samples, DNA was successfully extracted and quantified. The presence of grapevine DNA in these extractions was verified in 31 randomly selected samples by partially amplifying a Vitis psaA chloroplast gene. Then, genotyping with 13 SSR was performed in a laboratory where grapevine DNA had never been amplified. The success of the genotyping analyses varied, from a third of the samples producing no positive results, to others working well, like modern DNA samples. The genetic profiles obtained were then compared to those stored in international databases (ICVV and VIVC). The samples were also genotyped for 48 SNPs and confirmed the SSR results. Up to 32 of the herbarized samples were successfully identified as 19 different varieties from the Western Balkans and neighboring regions, such as cv. ‘Kadarka Kek’, indicating their continuous cultivation for more than 200 years. The combined analysis of ancient and modern samples allows elucidating the historical evolution of the crop in Serbia.
Acknowledgements: Ministry of Agriculture, Forestry, and Water Management, Republic of Serbia; and European COST Action CA17111 INTEGRAPE.
Issue: ICGWS 2023
1Instituto de Ciencias de la Vid y del Vino (ICVV, CSIC-CAR-UR), Departamento de Viticultura, Logroño, Spain
2Instituto de Historia, Departamento de Arqueología y Procesos Sociales, Madrid, Spain
3University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Novi Sad, Serbia
4Universidad de Santiago de Compostela, Santiago de Compostela, Spain
5University of Belgrade, Institute for Multidisciplinary Research, Belgrade, Serbia
6University of Belgrade, Faculty of Biology, Belgrade, Serbia
7University of Belgrade, Faculty of Agriculture, Department of Viticulture, Belgrade, Serbia
8University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia
9University of Tuebingen, Archaeo- and Palaeogenetics, Tuebingen, Germany
10Max-Plank Institute for Biology, Molecular Biology, Tuebingen, Germany
11University of Western Ontario, Department of Biology, London, Canada