Molecular Ecology (2006) 15, 3707–3714 doi: 10.1111/j.1365-294X.2006.03049.x MultipleBlackwell Publishing Ltd origins of cultivated grapevine (Vitis vinifera L. ssp. sativa) based on chloroplast DNA polymorphisms R. ARROYO-GARCÍA,*† L. RUIZ-GARCÍA,* L. BOLLING,* R. OCETE,‡ M. A. LÓPEZ,‡ C. ARNOLD,§ A. ERGUL,¶ G. SÖYLEMEZO˝LU,** H. I. UZUN,†† F. CABELLO,‡‡ J. IBÁÑEZ,‡‡ M. K. ARADHYA,§§ A. ATANASSOV,¶¶ I. ATANASSOV,¶¶ S. BALINT,*** J. L. CENIS,††† L. COSTANTINI,‡‡‡ S. GORIS- LAVETS,§§§ M. S. GRANDO,‡‡‡ B. Y. KLEIN,¶¶¶ P. E. M C GOVERN,**** D. MERDINOGLU,†††† I. PEJIC,‡‡‡‡ F. PELSY,†††† N. PRIMIKIRIOS,§§§§ V. RISOVANNAYA,§§§ K. A. ROUBELAKIS-ANGELAKIS,§§§§ H. SNOUSSI,¶¶¶¶ P. SOTIRI,***** S. TAMHANKAR,††††† P. THIS,‡‡‡‡‡ L. TROSHIN,§§§§§ J. M. MALPICA,† F. LEFORT¶¶¶¶¶ and J. M. MARTINEZ-ZAPATER* *Departamento de Genética Molecular de Plantas, Centro Nacional de Biotecnología, CSIC, C/Darwin 3, 28049 Madrid, Spain, †Departamento de Biotecnología, INIA, Ctra. de A Coruña km 7, 28040 Madrid, Spain, ‡Departamento de Fisiología y Zoología, Universidad de Sevilla, C/Reina Mercedes 6, 41012 Sevilla, Spain, §National Center of Competence in Research 'Plant Survival', University of Neuchâtel, Emile Argand 11, Ch 2007, Neuchâtel, Switzerland, ¶Institute of Biotechnology, Ankara University, 065000 Besevler-Ankara, Turkey, **Department of Horticulture, Faculty of Agriculture, Ankara University, 06110 Diskapi-Ankara, Turkey, ††Department of Horticulture, Faculty of Agriculture, Akdeniz University, Antalya, Turkey, ‡‡Finca El Encin, IMIDRA, Alcalá de Henares, 28800 Madrid, Spain, §§National Clonal Germplasm Repository, USDA-ARS, University of California, Davis, CA 95616, USA, ¶¶AgroBioInstitute, D. Tzankov 8, 1164 Sofia, Bulgaria, ***Department of Viticulture, University of Agricultural Sciences and Veterinary Medicine ‘Ion Ionescu de la Brad’, Aleea Mihail Sadoveanu nr. 3, 700490 Romania, †††IMIDA, La Alberca, 30150 Murcia, Spain, ‡‡‡Istituto Agrario S. Michele all’Adige, Via Mach 1, 38010 Trento, Italy, §§§Institute of Vine and Wine Magarach, 31 Kirov St., 98600 Yalta, Crimea, Ukraine, ¶¶¶Hebrew University Hospital, Ein Kerem, Jerusalem, POB 12000, Israel, ****MASCA, University of Pennsylvania Museum, Philadelphia, PA 19104, USA, ††††UMR 1131 Santé de la Vigne et Qualité du Vin, INRA-ULP, 28 rue de Herrlisheim, BP 20507, 68021 Colmar, France, ‡‡‡‡Department of Plant Breeding, Genetics and Biometrics, University of Zagreb, HR-10000 Zagreb, Croatia, §§§§Department of Biology, University of Crete, 71409 Heraklion, Crete, Greece, ¶¶¶¶Laboratoire de Biotechnologie, Institut National de la Recherche Agronomique de Tunisie, Rue Hédi Karray, 2049 Ariana, Tunisia, *****Institute of Biology, Academy of Science, Tirana, Albania, †††††Agharkar Research Institute, Department of Science and Technology, Pune-411 004, India, ‡‡‡‡‡UMR DGPC Génétique de la Vigne, ENSAM-INRA 2pl. P. Viala, 34060 Montpellier, France, §§§§§Viticulture Department, Kuban State Agrarian University, 350044 Krasnodar, Russia, ¶¶¶¶¶Laboratory of Applied Genetics, School of Engineering of Lullier, University of Applied Sciences of Western Switzerland, 150 route de Presinge, 1254 Jussy, Switzerland Abstract The domestication of the Eurasian grape (Vitis vinifera ssp. sativa) from its wild ancestor (Vitis vinifera ssp. sylvestris) has long been claimed to have occurred in Transcaucasia where its greatest genetic diversity is found and where very early archaeological evidence, including grape pips and artefacts of a ‘wine culture’, have been excavated. Whether from Transcaucasia or the nearby Taurus or Zagros Mountains, it is hypothesized that this wine culture spread southwards and eventually westwards around the Mediterranean basin, together with the transplantation of cultivated grape cuttings. However, the existence of morphological differentiation between cultivars from eastern and western ends of the modern distribution of the Eurasian grape suggests the existence of different genetic contribution from local sylvestris populations or multilocal selection and domestication of sylvestris genotypes. To tackle this issue, we analysed chlorotype variation and distribution in 1201 samples of sylvestris and sativa genotypes from the whole area of the species’ distribution and studied their genetic relationships. The results suggest the existence of at least two important origins for the cultivated germplasm, one in the Near East and another in the western Mediterranean region, the latter of which gave rise to many of the current Western European cultivars. Indeed, over 70% of the Iberian Peninsula cultivars display chlorotypes that are only compatible with their having derived from western sylvestris populations. Correspondence: José Miguel Martínez Zapater, Fax: 34-915854506; E-mail: [email protected] © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd 3708 R. ARROYO-GARCÍA ET AL. Keywords: chloroplasts diversity, crop origins, domestication centres, domestication, genetic variation, grapevine Received 7 December 2005; revision received 12 May 2006; accepted 2 June 2006 c. 2200 bc, and on the coasts of the Italian and Iberian Introduction Peninsulas (c. 800 bc) (McGovern 2003). On the other hand, The Eurasian grape (Vitis vinifera L.) is the most widely the existence of morphological differentiation among cultivated and economically important fruit crop in the cultivars from distinct geographical areas in the Near East world (Vivier & Pretorius 2002). Cultivated grapevines and in the western Mediterranean region supports the (Vitis vinifera spp. sativa) are thought to have been second possibility in which wild local sylvestris germplasm domesticated from wild populations of Vitis vinifera spp. significantly contributed to the generation of grape cultivars, sylvestris (Levadoux 1956). These wild vines are dioecious possibly through multiple domestication events (Negrul plants still occurring in small isolated populations along 1938; Levadoux 1956; Mullins et al. 1992). This possibility is riverbank forests from the Atlantic coast of Europe to compatible with an eastern ancestral origin for the wine Tajikistan and the western Himalayas (Zohary & Hopf culture and viticulture practices and its spread from east 2000). The exploitation of wild grape fruit as a food source to west. Resolving this issue has important implications by Palaeolithic hunter-gatherer populations is well for understanding the origin of current grape cultivars documented at many prehistoric sites across Europe and provides information on the processes involved in (Zohary 1996). However, the domestication of the grapevine the domestication of woody plant species. has been linked to the production of wine, which required The analysis of the amount and distribution of genetic more fruit to be viable as well as storage containers made variation in cultivated (V. vinifera ssp. sativa) and wild of pottery to preserve the beverage, developments that (V. vinifera ssp. sylvestris) populations can help in under- did not occur until the Neolithic period (c. 8500–4000 bc) standing the process of grapevine domestication. V. vinifera (McGovern et al. 1986). The domestication process involved plants are highly heterozygous and the vegetative propa- the selection of hermaphrodite genotypes producing larger gation of cultivars has maintained their high heterozygosity and sweeter berries of attractive colours and the development levels. When cultivars from the same geographic regions of techniques for their vegetative propagation (Zohary & are grouped, nuclear DNA microsatellite markers pro- Hopf 2000). vide weak discrimination between different geographic Major questions regarding grapevine domestication groups, with the greatest variation existing within the cultivar concern the number of domestication events and the groups themselves (Aradhya et al. 2003). Additionally, geographic locations where they took place. Two basic European grape cultivars have a complex history of move- divergent hypotheses can be formulated: (i) a restricted ment over growing regions, which hampers the recogni- origin hypothesis in which domestication took place tion of clear geographic trends in their distribution from a limited wild stock in a single location, with those (Sefc et al. 2000). To try overcoming these problems we cultivars subsequently being transplanted to other regions have used variation in the chloroplast genome to analyse (Olmo 1976); and (ii) a multiple-origin hypothesis in which the relationships between sylvestris and sativa grapevine domestication could have involved a large number of groups. The chloroplast genome has a lower evolutionary founders recruited during an extended time period and rate than the nuclear genome (Provan et al. 1999), and along the entire distribution range of the wild progenitor being maternally inherited in grapevine (Arroyo-García species (Mullins et al. 1992). In agreement with the first et al. 2002) can only be disseminated by seeds or cuttings. possibility, archaeological research has traced the earliest Furthermore, preliminary genotyping of grapevine evidence for large-scale winemaking, presumably exploiting cultivars from different locations for a few chloroplast a domesticated plant, to the Neolithic period in the northern microsatellite loci have shown the existence of a reduced mountainous regions of the Near East, encompassing the number of chlorotypes, which could show specific geographic northern Zagros, eastern Taurus and Caucasus Mountains distributions (Arroyo-García