Combining Microsatellite Markers and Ampelography for Better Management of Romanian Grapevine Germplasm Collections

Aiming to investigate the correspondence of phenotypic variability and molecular markers, 50 varieties of Vitis vinifera ssp. vinifera, ancient grapevine varieties grown on Romanian territory, were selected to be analysed. All varieties were subjected to ampelographic analyses with OIV descriptors and also to molecular analyses with 13 microsatellites. The morphology description with recommended methods corresponded with the reference literature, proving the correct registration of the varieties at the moment of acquisition. Markers efficiency for assessing the genetic diversity among studied varieties was evaluated by computation of statistic parameters referring to the polymorphisms found. Among the internationally recommended SSR markers, ISV4 and VMCNG4b9 markers displayed interesting PIC and PI values. By comparing the SSR profiles of the 50 Romanian grapevine varieties with those in the Eu database, the following can be concluded: confirmation of 10 synonymies mentioned in old documents, while 3 synonymies mentioned in old reference literature were not confirmed, new synonyms were found for 4 accessions, additional synonyms were found for 4 accessions, which are to be added to the previous ones already mentioned in the literature, the synonymies in three groups of accessions were documented for the first time in  the specialised literature, the unicity of three Romanian accessions was proved (ʻMoroştinăʼ, ʻNegru mareʼ and ʻRomânieʼ) and the genetic profiles of 15 varieties were identical to those already recorded in the investigated database

Genotyping of Vitis vinifera L. within the Slovak national collection of genetic resources

Abstract Microsatellites were used as a very effective tool for genetic diversity analysis and characterization of 51 grapevine (Vitis vinifera L.) accessions from the national collection of genetic resources. Genetic diversity was relatively high, 8.91 alleles were detected per analysed microsatellite locus in average, and fifty-one accessions were distinguished into 45 groups. Distribution of recent Slovak cultivars across the dendrogram accented both their genetic diversity and the effectiveness of the national breeding program in maintaining genetic diversity and generating new genetic variants. Each cultivar was different from the others and twelve of them contained 77.6% of the total genetic diversity of the whole analysed set. Microsatellite patterns were also able to confirm parentage in selected Slovak cultivars. An unusual phenomenon of triallelism was also detected in one of the analysed accessions. The present study has initiated molecular characterization within the national grapevine genetic resource collection and their comparison with well-established international cultivars

A major QTL is associated with berry grape texture characteristics

9restrictedInternationalItalian coauthor/editorBerry texture and berry skin mechanical properties are traits with high agronomic relevance because they are related to quality parameters and marketing requirements of wine, table, and raisin grapes. Searching for QTLs linked to berry texture, an F1 population of 152 individuals and their parents were used in this study. These F1 plants were obtained crossing Raboso Veronese, a seeded black wine grape cultivar, and Sultanina, a seedless white grape variety, especially used for raisins. Density flotation was applied for berry sorting improving the management of many and highly variable genotypes, irrespective of the quantification of specific molecule classes. Berries were evaluated for technological ripeness parameters and mechanical properties. Texture parameters were taken as raw data and as data normalised on berry dimensions, i.e., berry diameter or surface or volume. SSR molecular markers were used to produce a genetic map and a major QTL for berry texture was found on chromosome 18 with traits related to berry firmness showing a phenotypical explained variance higher than 60 %, and traits related to berry resilience, springiness and cohesiveness showing a variance higher than 50 %. Surprisingly, this QTL showed to be associated with SSR markers linked to VviAGL11, the main gene linked to seedlessness. VviAGL11 expression and co-expression profiling during grape ripening was evaluated using available information; this data suggested a role for this gene on the texture of a ripe berry.restrictedCrespan, Manna; Migliaro, Daniele; Vezzulli, Silvia; Zenoni, Sara; Tornielli, Giovanni Battista; Giacosa, Simone; Paissoni, Maria Alessandra; Río Segade, Susana; Rolle, LucaCrespan, M.; Migliaro, D.; Vezzulli, S.; Zenoni, S.; Tornielli, G.B.; Giacosa, S.; Paissoni, M.A.; Río Segade, S.; Rolle, L

Parentage Atlas of Italian Grapevine Varieties as Inferred From SNP Genotyping

The Italian grape germplasm is characterized by a high level of richness in terms of varieties number, with nearly 600 wine grape varieties listed in the Italian National Register of Grapevine Varieties and with a plethora of autochthonous grapes. In the present study an extended SNP genotyping has been carried out on Italian germplasm of cultivated Vitis vinifera subsp. sativa and Vitis hybrids. Several hundred Italian varieties maintained in the repositories of scientific Institutions and about one thousand additional varieties derived from previous studies on European, Southern Italy, Magna Graecia and Georgian germplasm were considered. The large genotyping data obtained were used to check the presence of homonyms and synonyms, determine parental relationships, and identify the main ancestors of traditional Italian cultivars and closely-related accessions. The parentage among a set of 1,232 unique varieties has been assessed. A total of 92 new parent-offspring (PO) pairs and 14 new PO trios were identified. The resulted parentage network suggested that the traditional Italian grapevine germplasm originates largely from a few central varieties geographically distributed into several areas of genetic influence: “Strinto porcino” and its offspring “Sangiovese”, “Mantonico bianco” and “Aglianico” mainly as founder varieties of South-Western Italy (IT-SW); Italian Adriatic Coast (IT-AC); and Central Italy with most varieties being offsprings of “Visparola”, “Garganega” and “Bombino bianco”; “Termarina (Sciaccarello)” “Orsolina” and “Uva Tosca” as the main varieties of North-Western Italy (IT-NW) and Central Italy. The pedigree reconstruction by full-sib and second-degree relationships highlighted the key role of some cultivars, and, in particular, the centrality of “Visparola” in the origin of Italian germplasm appeared clear. An hypothetical migration of this variety within the Italian Peninsula from South to North along the eastern side, as well as of “Sangiovese” from South to Central Italy along the Western side might be supposed. Moreover, it was also highlighted that, among the main founders of muscat varieties, “Moscato bianco” and “Zibibbo (Muscat of Alexandria)” have spread over the whole Italy, with a high contribution by the former to germplasm of the North-Western of the peninsula

Grapevine (Vitis vinifera L.) varietal assortment and evolution in the Marche region (central Italy)

14openInternationalItalian coauthor/editorItaly 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.openCrespan, Manna; Migliaro, Daniele; Larger, Simone; Pindo, Massimo; Palmisano, Massimo; Manni, Alvise; Manni, Elisa; Polidori, Enzo; Sbaffi, Francesco; Silvestri, Quintilio; Silvestroni, Oriana; Velasco, Riccardo; Virgili, Settimio; Camilli, GiuseppeCrespan, M.; Migliaro, D.; Larger, S.; Pindo, M.; Palmisano, M.; Manni, A.; Manni, E.; Polidori, E.; Sbaffi, F.; Silvestri, Q.; Silvestroni, O.; Velasco, R.; Virgili, S.; Camilli, G

Integrated Bayesian Approaches Shed Light on the Dissemination Routes of the Eurasian Grapevine Germplasm

The domestication and spreading of grapevine as well as the gene flow history had been described in many studies. We used a high-quality 7k SNP dataset of 1,038 Eurasian grape varieties with unique profiles to assess the population genetic diversity, structure, and relatedness, and to infer the most likely migration events. Comparisons of putative scenarios of gene flow throughout Europe from Caucasus helped to fit the more reliable migration routes around the Mediterranean Basin. Approximate Bayesian computation (ABC) approach made possible to provide a response to several questions so far remaining unsolved. Firstly, the assessment of genetic diversity and population structure within a well-covered dataset of ancient Italian varieties suggested the different histories between the Northern and Southern Italian grapevines. Moreover, Italian genotypes were shown to be distinguishable from all the other Eurasian populations for the first time. The entire Eurasian panel confirmed the east-to-west gene flow, highlighting the Greek role as a “bridge” between the Western and Eastern Eurasia. Portuguese germplasm showed a greater proximity to French varieties than the Spanish ones, thus being the main route for gene flow from Iberian Peninsula to Central Europe. Our findings reconciled genetic and archaeological data for one of the most cultivated and fascinating crops in the world

Novel loci associated with resistance to downy and powdery mildew in grapevine

Among the main challenges in current viticulture, there is the increasing demand for sustainability in the protection from fungal diseases, such as downy mildew (DM) and powdery mildew (PM). Breeding disease-resistant grapevine varieties is a key strategy for better managing fungicide inputs. This study explores the diversity of grapevine germplasm (cultivated and wild) from Caucasus and neighboring areas to identify genotypes resistant to DM and PM, based on 13 Simple Sequence Repeat (SSR) loci and phenotypical (artificial pathogen inoculation) analysis, and to identify loci associated with DM and PM resistance, via Genome-Wide Association Analysis (GWAS) on Single Nucleotide Polymorphism (SNP) profiles. SSR analysis revealed resistant alleles for 16 out of 88 genotypes. Phenotypic data identified seven DM and 31 PM resistant genotypes. GWAS identified two new loci associated with DM resistance, located on chromosome 15 and 16 (designated as Rpv36 and Rpv37), and two with PM resistance, located on chromosome 6 and 17 (designated as Ren14 and Ren15). The four novel loci identified genomic regions rich in genes related to biotic stress response, such as genes involved in pathogen recognition, signal transduction and resistance response. This study highlights potential candidate genes associated with resistance to DM and PM, providing valuable insights for breeding programs for resistant varieties. To optimize their utilization, further functional characterization studies are recommended

Clones Identification and Genetic Characterization of Garnacha Grapevine by Means of Different PCR-Derived Marker Systems

This study uses PCR-derived marker systems to investigate the extent and distribution of genetic variability of 53 Garnacha accessions coming from Italy, France and Spain. The samples studied include 28 Italian accessions (named Tocai rosso in Vicenza area; Alicante in Sicily and Elba island; Gamay perugino in Perugia province; Cannonau in Sardinia), 19 Spanish accessions of different types (named Garnacha tinta, Garnacha blanca, Garnacha peluda, Garnacha roja, Garnacha erguida, Garnacha roya) and 6 French accessions (named Grenache and Grenache noir). In order to verify the varietal identity of the samples, analyses based on 14 simple sequence repeat (SSR) loci were performed. The presence of an additional allele at ISV3 locus (151 bp) was found in four Tocai rosso accessions and in a Sardinian Cannonau clone, that are, incidentally, chimeras. In addition to microsatellite analysis, intravarietal variability study was performed using AFLP, SAMPL and M-AFLP molecular markers. AFLPs could discriminate among several Garnacha samples; SAMPLs allowed distinguishing few genotypes on the basis of their geographic origin, whereas M-AFLPs revealed plant-specific markers, differentiating all accessions. Italian samples showed the greatest variability among themselves, especially on the basis of their different provenance, while Spanish samples were the most similar, in spite of their morphological diversity