Gábor Gyulai 1, Zoltán Tóth 1, Zoltán Szabó 1, Ferenc Gyulai 2, Richárd Lágler 1, László Kocsis 3 and László Heszky 1 Domestication Events of Grape (Vitis vinifera) from Antiquity and the Middle Ages in Hungary from Growers’ Viewpoint Morphological reconstruction of Acareosperma ; Ampelocissus ¸ in southwest Asia and southern ancient grapes ( Vitis vinifera) Ampelopsis (pepper-vine s); Cay- Transcaucasia (Armenia and based on archaeological seed ratia ; Cissus (treebine s); Clema- Georgia). Seeds of Vitaceae are remains provide insight into the ticissus ; Cyphostemma ; Leea ; easily identified from a suite of domestication and cultivation Muscadinia ; Nothocissus ; Parthe- unique and distinctive morpho- events of grapes in Hungary. nocissus ; Pterisanthes ; Ptero- logical characters (particularly a Ancient grape seeds were cissus ; Rhoicissus ; Tetrastigma ; pair of ventral in folds and a dorsal excavated at Roman and Medieval Vitis (grape s); and Yua of about chalazal scar). archaeological sites in Hungary 700 species (Facsar 1970; Terpó The wild, dioecious ancestor and analyzed by LM (Light 1976). The genus Vitis consists of form of V. vinifera ssp. silvestris Microscopy) and SEM (Scanning about 60 inter-fertile species (syn.: V. silvestris ) still co exists Electron Microscopy). Excavation including about fifteen species of with the cultivated, hermaphrodite sites included Budapest ( Aquin- agronomic importance (Table 1). flower form of V. vinifera ssp. cum ; 2 nd –4th CENT . A.D. Of them, V. vinifera (2n = 4× = 38) vinifera (syn. V. vinifera ) in Hungary) and Keszthely (Fenék- is the only species which is Eurasia and North Africa (This et puszta) of Roman Age (5 th CENT . indigenous to Eurasia, with a al . 2006). Today, thousands of A.D., Hungary); and Gyır (Ece; relatively small nuclear (nuDNA) cultivars have been developed 11 –12 th CENT . A.D., Hungary), genome size of 0.475 –0.5×10 9 which are generally classified in Debrecen (13 th CENT . A.D., DNA base pair (bp); and a 160,928 three main groups according to Hungary) and the King ’s Palace of bp of chloroplast cpDNA (Jansen their final production, as wine the Árpád Dynasty at the Castle of et al. 2006) and a regular size of grapes, table grapes including Buda, Budapest (15 th CENT . A.D., higher plant mtDNA (1 –400,000 modern seedless grapes, and Hungary) of the Middle Ages. bp) . Most genera of family raisins. Ancient seeds were compared to Vitaceae have 2n = 38 chromoso- Genetically, dioecy in wild thirty current grape varieties of mes (n = 19), but species of grape is encoded by a single gene; similar seed size, shape, and Muscadinia , Ampelocissus , female individuals are homo- morphology. The modern grape Parthenocissus , and Ampelopsis gametic carrying homozygous variety Vitis vinifera cv. ‘kék have 2n = 40 (n = 20) recessive pistil- su ppressor alleles bakator ’ ( syn .: ‘ Blue Bocca chromosomes, and species of the (su msu m) which suppress the d’Oro ’; ‘ aranybogyó ’) was found genus Cissus has 2n = 24 (n = 12) development of anthers (and most similar in seed morphology chromosomes. pollen). Male plants are hetero- to one of the ancient samples (15 th Changes in seed ( ‘pip ’) shape, zygous (SU Fsu m) carrying a CENT. Debrecen, Hungary) which wild grapes have rounder pips with dominant pistil-suppressing Su F indicates the antiquity of this short beaks, while seeds of allele. cultivar. cultivated grape tend to be more The shift, under domestication, elongated with longer beaks. Seed to bisexual (hermaphrodism) The genus Vitis morphology indicates that flowers took place via a single + domestication of grape ( Vitis mutation to SU m which is also Species of the plant family vinifera ) began with the Eurasian dominant over su resulting in two Vitaceae are woody climbers wild grape ( V. sylvestris) about genotypes of hermaphroditc grape comprising 13 –17 genera: 5,500 –5,000 B .P. (before present) types (SU +su m and SU +SU +). 1,2 Szent István University, 1Institute of Genetics and Plant Breeding, and 2KTI, Gödöllô, Hungary; 3Pannon University, Faculty of Georgikon, Keszthely, Hungary. e-mail: [email protected] 8 Hungarian Agricultural Research 2009/3 –4 Domestication events in the Table 1. Vitis species (1 –27), hybrids (1 –9) and gene bank samples genus Vitis (1 –12) The oldest (8,400 B .P.) wild Vitis species Vitis hybrids* Vitis gene bank samples 1. Vitis acerifolia 1. V. arizonica x V. rupestris 1. Vitis sp. grape ( Vitis sylvestris ) seeds 2. Vitis aestivalis 2. V. berlandieri x V. riparia 2. Vitis sp. 196-17 (about 3 mm long) were excavated 3. Vitis amurensis 3. V. berlandieri x V. 3. Vitis sp. 216-N in Turkey, at Nevali Çori (NÇ) rupestris 4. Vitis arizonica 4. V. berlandieri x V. vinifera 4. Vitis sp. 44-53M located near the Turkish city of 5. Vitis bashanica 5. V. cinerea x V. riparia 5. Vitis sp. 8007 Urfa (37 °60’N, 38 °70’E, 490 m 6. Vitis berlandieri 6. V. cinerea x V. rupestris 6. Vitis sp. 8658 above sea level) on the slope of a 7. Vitis betulifolia 7. V. labrusca x V. vinifera 7. Vitis sp. cv. 'Norton' Euphrates side valley, Hilvan 8. Vitis bryoniifolia 8. V. pseudoreticulata x V. 8. Vitis sp. CWD 96.701 vinifera province. The first convincing 9. Vitis cinerea (downy grape) 9. V. riparia x V. rupestris 9. Vitis sp. Nie 372 evidence of Vitis vinifera seeds 10. Vitis davidii 10. Vitis sp. Nie 415 with indications of grape culti- 11. Vitis flexuosa * Interspecific hybrids registered in 11. Vitis sp. NL- Hungary (2006): vation were also uncovered in 12. Vitis heyneana 12. Vitis sp. Qiu 13. Vitis kelungnsis Turkey at Kurban Höyük (5.700 – ‘Bianka’; ‘Csillám’; 14. Vitis labrusca(Concord grape) ‘Duna gyöngye’; ‘Esther’; 5.200 B.P. non-calibrated radio- 15. Vitis piasezkii ‘Fanny’; ‘Göcseji zamatos’; 16. Vitis popenoei (totoloche grape) ‘Kunleány’; ‘Medina; carbon time), followed by the early ‘Nero’; ‘Odysseus’; 17. Vitis pseudoreticulata ‘Orpheus’; ‘Platina’; Bronze Age samples (3,200 –1900 18. Vitis quinquangularis ‘Pannon frankos’; ‘Pölöskei muskotály’; 19. Vitis riparia (riverbank grape) ‘Refrén’; ‘Taurus’; B.P.) along the Jordan Valley, at 20. Vitis rotundifolia (fox grape) ‘Teréz’; ‘Viktória gyöngye’; ‘Zalagyöngye’. Tell Shuna (Jordan; Chalcoitic), 21. Vitis rupestris (rock grape) 22. Vitis shuttleworthii (callose Jericho (Cisjordan; early Bronze 23. Vitis sinocinerea Age), and Arad (Israel, early 24. Vitis thunbergii 25. Vitis tiliifolia Bronze Age) (Jacquat and 26. Vitis vinifera(wine grape) Martinoli 1999). Ancient grape 27. Vitis yeshanensis seeds were also excavated at Semma (Sudan) 3,500 B .P. Zheng He (1405 –1435), or growing in North America. Some The earliest evidence of wine Columbus voyages (first: Aug. 3 of these hybrids became resistant production (jars from Godin) was 1492 to March 15 1493; second: to Phyloxera (an insect pest), found in Iran (Hajji Firuz Tepe site Sept. 25 1493 to June 11 1495; which devastated European in the Zagros Mountains) about third: May 30 1498 to Nov. 15 vineyards in the 1880s, and 7,400 –7,000 B .P. (This et al . 1500; fourth: May 11 1502 to Nov. supplied resistant rootstocks for 2006) and 5,500 4,900 B .P. Greek, 7 1504). The first plantations in replantations. This event indicates Latin, and Egypt vine amphoras North America were established that the diversity of grape genome with gelyfied vine remains were on the West Coast by Spanish has been narrowed twice; first by found in the hulls of sunken ships missionaries and later by Hun- the Biblical sunk, similar to the famous garian viticulturists like Ágoston flood, followed by the re shipwreck remains at Uluburun Haraszty who is considered the plantation of Noah ‘the first near Kas (Turkey). Grape ‘father of California’s grape- vintner ’ (Genesis 9) on Mount cultivation gradually spread to growing industry ’. Haraszty Ararat, and second by Phyloxera Mesopotamia, Assyria, and Egypt imported 200,000 grape cuttings (This et al. 2002). Unlike the (about 5, 500–5,000 B.P.), and from Europe from 1849, including genome for dioecious V. sylvestris , further west along the grape varieties from his native genetic diversity of grape has been Mediterranean to Phoenicia, Hungary. With the passing of time, narrowing continuously as the Greece, North Africa and then to Haraszty developed over half a result of vegetative propagation the entire Roman Empire north to million California acres to either by rooting of twigs, or by Pannonia (Hungary) and German viticulture, making wine growing grafting. tribes. Viticulture also spread second to orange production in the In Hungary, the earliest wild eastward along the Silk Road and state’s agricultural economy. In grape ( Vitis sylvestris ) seed it reached China and Japan in recognition of his merits, Haraszty remains were found at Tiszapolgár 3,200 B .P. was named California’s State (5,300 B .C.) and the earliest Vitis Grape were introduced to the Commissioner of Viticulture (Sisa vinifera at Sopron (1,300 B .C.), Americas by European colonists 2006). which dates the origins of grape starting from the 16 th CENT. after European grape formed cultivations to the late Bronze Age either the early Chinese explorer hybrids with native Vitis species (Table 2, Fig. 1). Hungarian Agricultural Research 2009/3 –4 9 The earliest wine residue in Table 2. Vitis (V. sylvestris, V. vinifera and V. sp.) and wine remains Hungary dates back to 700 B .C. (at (pieces #) excavated in Hungary Fehérvárcsurgó), which places the beginnings of wine making to the Iron Age. Thus, there is evidence that both grape cultivation and wine making date to well before to the Roman period in Hungary. How to recover ancient DNA Excavated and wet-sieved sediment samples of the study presented were processed by flotation followed by seed sorting and identification in the laboratory according to Gyulai et al . (2001, 2006).