Vitis Vinifera L.) Varietal Assortment and Evolution in the Marche Region (Central Italy
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Received: 25 February 2021 y Accepted: 23 June 2021 y Published: 9 July 2021 DOI:10.20870/oeno-one.2021.55.3.4628 Grapevine (Vitis vinifera L.) varietal assortment and evolution in the Marche region (central Italy) Manna Crespan1*, Daniele Migliaro1, Simone Larger2, Massimo Pindo2, Massimo Palmisano3, Alvise Manni4, Elisa Manni5, Enzo Polidori6, Francesco Sbaffi5, Quintilio Silvestri6, Oriana Silvestroni7, Riccardo Velasco1, Settimio Virgili6 and Giuseppe Camilli6 1 CREA Research Centre of Viticulture and Enology, 31015 Conegliano, Treviso, Italy 2 Unit of Computational Biology, Research and Innovation Centre, Fondazione Edmund Mach, 38010 San Michele all’Adige, Trento, Italy 3 Department of Engineering and Architecture, University of Trieste, 34127 Trieste, Italy 4 Marche history scholar, 62012 Civitanova Marche, Italy 5 I.I.S., Istituto Istruzione Superiore «Morea Vivarelli», 60044 Fabriano, Ancona, Italy 6 ASSAM, Agenzia Servizi Settore Agroalimentare Marche, 60027 Osimo, Ancona, Italy 7 Università Politecnica delle Marche, Italy, Dipartimento di Scienze Agrarie, Alimentari e Ambientali, Ancona, Italy *corresponding author: [email protected] Associate editor: Mario PezzottiABST ABSTRACT Italy is one of the richest countries in terms of grapevine varietal assortment, and the Marche region in central Italy has an ancient winegrowing tradition. Increasing interest in autochthonous grapevine varieties prompted efforts to recover and identify local minor germplasm also in this region, and to search for pedigree relationships and determine the evolution of varietal assortment. In the present study this was done using nuclear and chloroplast microsatellite markers and SNP markers from the Vitis18kSNP chip. Eighteen new genotypes were found, of which seventeen belonged to well-known, less known and even unknown vines found to be specific to the Marche region. The rearranged pedigree highlighted the complex intertwining relationships between Marche varieties. Second-degree relationships were also derived. Some minor Marche varieties have increased the number of parent-offspring related members of Garganega, Sangiovese, Crepolino/Visparola and Sciaccarello, which are varieties already recognised as founders or recurrent parents of many Italian cultivars. Crepolino/Visparola was shown to be a key variety in the evolution of the Marche varietal assortment, having played the role of parent and grandparent, as explained by the presence of this genotype in the region in ancient times. Surprisingly, Semidano, a well-known Sardinian variety, played a clear role as parent or grandparent of three minor Marche varieties, testifying to its presence in central Italy in ancient times. Incrocio Bruni 54 was confirmed as an offspring of Sauvignon and Verdicchio, as declared by the breeder. The parentage of Lacrima was completed: this variety was found to be a spontaneous cross between Nera Rada and Aleatico. A dendrogram of genetic similarity clearly resembled the groups defined with the pedigree reconstruction and it gave an indication of the genetic similarity of the varieties excluded from the strictest parentage links. ACT KEYWORDS Italian grapevine germplasm, variety identification, old autochthonous varieties, pedigree relationships, microsatellite markers (SSR markers), Single Nucleotide Polymorphism (SNP markers) OENO One 2021, 3, 17-37 © 2021 International Viticulture and Enology Society - IVES 17 Manna Crespan et al. INTRODUCTION resources for their conservation, characterisation and potential future exploitation. Over the period Italy is one of the richest countries in terms of 1990-2012, some local, national and international grapevine varietal assortment, with almost 600 grapevine varieties were evaluated and the most cultivars used in wine production registered interesting ones were registered in the Italian in the Italian National Catalogue of Grapevine National Catalogue of Grapevine Varieties, like Varieties (http://catalogoviti.politicheagricole.it). Vernaccia nera grossa in 2008 and Garofanata in The origin of the cultivated varieties can be 2013 (http://catalogoviti.politicheagricole.it). In determined by reconstructing their kinship; 2014, a larger recovery programme was set up: indeed, in the wine supply chain, describing a more than one hundred grapevine samples were wine through the synergy between cultivation, collected in the Marche region and genotyped with quality and cultural issues is becoming microsatellite (SSR) markers to speed up their increasingly important. The Marche region in identification. Indeed, classical ampelographic central Italy and overlooking the Adriatic Sea identification is often difficult due to high varietal has an ancient winegrowing tradition. In light of variability, often combined with poor vine health. increasing consumer interest in local products, Later, rare and unexplored genetic resources were research on local grapevine germplasm and its preserved in a dedicated germplasm repository; conservation and exploitation play an important a search for verbal and written information on role in supporting the economic sustainability of these poorly known grapevines was also made. farms. Viticulture is an important sector in the Pedigree studies were then implemented on Marche region, with approximately 17,300 ha varieties cultivated in the Marche region, in of vineyards and wine production of around one which 12 nuclear and 8 chloroplast SSRs were million hectolitres (https://www.assoenologi.it/ combined with SNPs from the Vitis18kSNP array. wp-content/uploads/2020/09/report_previsioni_ The very large number of point mutations in the vendemmiali.pdf). Even if fifty-six grapevine grapevine genome make SNP (Single Nucleotide varieties are suitable for cultivation in the Marche, Polymorphism) markers a powerful tool with only six are grown in over 70 % of the region’s many applications. Next generation sequencing vineyards, namely Sangiovese, Montepulciano, provided hundreds of thousands of SNP markers Verdicchio, Trebbiano Toscano/Biancame, in the whole Vitis spp. genome. A first Vitis chip Passerina and Pecorino. The first two varieties are array, the Vitis9kSNP, was developed in 2010 used to produce the renowned Rosso Piceno wines (Myles et al., 2010), with 9,000 SNP detected and the third the Verdicchio dei Castelli di Jesi in 11 Vitis vinifera L. varieties and six wild Vitis wines; together, these three varieties account for species. The GrapeReSeq Consortium developed more than 50 % of the Marche appellation wines. a new one, the Vitis18kSNP, obtaining around 18,000 SNPs from 47 V. vinifera varieties, Significant efforts have been made to recover 12 wild Vitis species and 5 Muscadinia rotundifolia and identify local minor grapevine germplasm varieties (Le Paslier et al., 2013). This tool is used in many Italian regions, like Apulia, Campania, in pedigree studies as it contributes to finding new Emilia Romagna, Friuli Venezia Giulia, Liguria, relationships, and it confirms or questions those Sardinia, Sicily, Tuscany (Costantini et al., 2005; found using only SSRs (Myles et al., 2011; Ruffa De Mattia et al., 2007; Torello Marinoni et al., et al., 2016; Laucou et al., 2018; De Lorenzis 2009; Carimi et al., 2010; Crespan et al., 2011; et al., 2019; Crespan et al., 2020; Raimondi De Lorenzis et al., 2014; Schneider et al., 2014; et al., 2020; D’Onofrio et al., 2021). Indeed, Mercati et al., 2016; D’Onofrio et al., 2016; the Vitis18kSNP surpasses SSRs, because it can Gristina et al., 2017; Pastore et al., 2020), and provide information on thousands of points in the variety pedigrees have been reconstructed for the genome in just one analysis, making up for their most widespread and renowned ones and for those very low polymorphism. having local importance, helping to unravel the intricate skein of the structure of Italian grapevine MATERIALS AND METHODS populations (Di Vecchi-Staraz et al., 2007; Ruffa 1. Plant material obtained during a sampling et al., 2016; Crespan et al., 2020; Raimondi campaign in the Marche region, nSSR et al., 2020; D’Onofrio et al., 2021). Similar efforts genotyping and varietal identification have been made in the Marche region, because the resulting knowledge is very useful in science and One hundred and twenty-two vines were sampled has cultural and marketing appeal. Since 1990, in the Marche region and preserved in the ASSAM ASSAM has recovered local grapevine genetic repository (Table 1); the sampling sites are 18 © 2021 International Viticulture and Enology Society - IVES OENO One 2021, 3, 17-37 FIGURE 1. Sites of recovery in Marche region of the 18 grapevine varieties lacking the SSR profile in the VIVC, which are not yet enrolled in the Italian catalogue and in bold in Table 1. Circle colours correspond to accession berry colour, black (red circles) or white (yellow circles). indicated when available and reported in Figure 1 genotyped samples were identified using the CREA for the accessions not listed or lacking an SSR Viticulture and Enology molecular database, the profile in the Vitis International Variety Catalogue available literature and the Vitis International (VIVC), or for those not enrolled in the Italian Variety Catalogue (VIVC, http://www.vivc.de). catalogue. 2. Plant material selected for pedigree study Often these vines did not have a varietal name and thus generic names were used for the samples, A selection of sixty-seven grapevine varieties referring, for example, to a vine trait, like grape from two Italian repositories (CREA Viticulture