SSR molecular marker analysis of the grapevine germplasm of Montenegro

摘要

Aim: Given that the information about the origin, genetic relationships, and diversity of Montenegrin grapevines is still partial, we performed a detailed analysis of the germplasm in this country using simple sequence repeat (SSR) markers. Our main goal was to determine the identity of cultivars unique to Montenegro and those shared with other countries, especially the neighbouring ones.Methods and results: Seventy samples were collected and 14 genotypes were found. After SSR profile comparison with available molecular databases and literature data, the identity of each genotype was established. Five well-known cultivars were found, the others being minor, lesser-known cultivars.Conclusion: This research provides an overview of the Montenegrin grapevine assortment. There are cultivars shared with other countries, mainly the neighbouring ones, while others are likely native to Montenegro. The Krato?ija population (alias Primitivo, Zinfandel and Crljenak Ka?telanski) has a large number of different names in Montenegro and also a wide morphological variability. Therefore, Montenegro is the best candidate as the origin and spreading point of this cultivar.Significance and impact of the study: The present study adds information on the identity, origin, diffusion and variability of some grapevine cultivars, allowing us to reconstruct the history and evolution of national and transnational ampelographic assortment of Montenegro. IntroductionMontenegro is a small country located in the Balkan Peninsula and overlooking the Adriatic Sea. Grapevine growing in Montenegro dates back before the Roman period (Vuksanovi?, 1977). The climatic, soil and orographic conditions of southern Montenegro define two wine regions: the Montenegro Skadar Lake basin, encompassing the Podgorica, Crmnica, Rijecki, Bjelopavlicki and Katunski sub regions, and the Montenegrin seacoast region, including the Boko-Kotorski, Budvansko-Barski, Ulcinjski and Grahovsko-Nudolski sub regions. Ancient and probably autochthonous cultivars dominate the present grapevine assortment of these two regions, and very heterogeneous populations appear as a consequence of perennial growing. Furthermore, there are numerous local grapevine cultivars lacking heterogeneity in their population, which have been registered as autochthonous even though they were not definitively identified. Most of these cultivars can be recognized using OIV codes, but it is not enough for their reliable identification (Uli?evi?, 1966; Pejovi?, 1988; Mara?, 2000).Current viticulture and wine production in Montenegro is based mainly on presumed autochthonous grapevine cultivars, such as Vranac, Krato?ija, Krsta?, and ?i?ak. The dominant cultivars for red wine production are Vranac and Krato?ija. Significantly less prevalent is Krsta?, used for white wine production, while no commercial wine is produced from ?i?ak. The presence of Mu?ka?ela and Trojka is significantly lower.The results of Calò et al. (2008) showed that Krato?ija is a synonym of the Californian Zinfandel, the Italian Primitivo and the Croatian Crljenak Ka?telanski, and suggested a first-degree relationship with the Montenegrin Vranac. Contrary to Vranac, which has stable morphological traits and has undergone expansion in the recent years, Krato?ija is very ancient and a lot of biotypes have been found within its population, not fully investigated yet. Because of its heterogeneity, Krato?ija is less present alone in the vineyards, and is found mostly in combination with Vranac, which dominates in the assortment of red grape cultivars. The heterogeneity of Krato?ija has been described by many authors (Stojanovi?, 1929; Buli?, 1949; Jelaska, 1954; Uli?evi?, 1966; Avramov, 1988; Pejovi?, 1988; Buri?, 1995; Bozinovik et al., 1998; Mara?, 2000; Mara? et al., 2004). It is well known that the features of an old grapevine cultivar may change with the accumulation and fixation of mutations in the shoot apical meristem, giving rise to many biotypes within the population, even with noticeable morphological or phenological differences. In this context, our work had different purposes. Using simple sequence repeat (SSR) markers, we wanted i) to investigate the identity of almost all grapevine cultivars grown in Montenegro; ii) to determine which cultivars are likely native to Montenegro and which are common to neighbouring countries; iii) to ascertain the identity of presumed Krato?ija biotypes collected in the experimental field of the Biotechnical Institute in Ljeskopolje and coming from different Montenegro private vineyards, where they are cultivated under many different names, depending on the area; and iv) to verify the correct identity of Vranac mother vines chosen for clonal selection process.Materials and methods1. Plant materialSeventy samples were selected for genotyping during the period from 2006 to 2010. The vines were collected from different viticultural areas of Montenegro (Table 1).Table 1. Accession list (name, pro