G. De Lorenzis

Azole-resistant Aspergillus fumigatus in the environment of northern Italy, May 2011 to June 2012

In recent years acquired azole resistance in Aspergillus fumigatus has been increasingly reported and a dominant mechanism of resistance (TR34/L98H) was found in clinical and environmental isolates. The aim of the present study was to investigate the prevalence of azole resistance in environmental A. fumigatus isolates collected in northern Italy. A. fumigatus grew from 29 of 47 soil samples analysed. Azole-resistant isolates were detected in 13% (6/47) of the soil samples and in 21% (6/29) of the soil samples containing A. fumigatus. High minimal inhibitory concentrations (MIC) of itraconazole (≥16 mg/L) and posaconazole (≥0.5 mg/L) were displayed by nine isolates from six different soil samples, namely apple orchard (1 sample), rose pot compost (2 samples), and cucurbit yields (3 samples). Seven isolates had a MIC=2 mg/L of voriconazole. Seven of nine itraconazole and posaconazole resistant isolates harboured the same TR34/L98H mutation of cyp51A. These findings, together with the occurrence of resistant clinical isolates, suggest that azole resistance should be considered in primary patient care.

Identification and characterization of grapevine genetic resources maintained in Eastern European Collections

Plenary lecture given at the 8th Mid-European Clay Conference, held in Kosice (Slovakia) on July 4-8th, 2016.COST MOLIM WG3 Meeting, Algorithm Development and High-performance Computing in Chemistry and Physics; Bratislava, March 21-22, 2016; http://web4.umb.sk/molim2016/Oral presentation given at the 8th Mid-European Clay Conference, held in Kosice (Slovakia) on July 4-8th, 2016. S6: application and treatment of clays and other industrial minerals.Cost Action Molim, Molecules in Motions, CM 1405; University Paris-Est Marne-La-Vallee, France, 27-29 August 2015This chapter provides an overview of the main biochemical transformations of major elements, including carbon, nitrogen, sulfur, iron and phosphorus in anaerobic digesters. Mineralization of organic matter during anaerobic digestion processes results in the production of inorganic carbonate, ammonium, sulfide, and phosphate species, which are involved in a complex network of chemical and biological reactions through interaction with available macro and micro nutrients as well as microbial processes with profound effects on the efficiency and stability of the anaerobic digester performance. The interplay of iron, phosphorus and sulfur cycles has recently attracted attention in the frame of research developed for the recovery of phosphorus on one hand and in the frame of the

Allelic variation in the VvMYBA1 and VvMYBA2 domestication genes in natural grapevine populations (Vitis vinifera subsp. sylvestris)

Grape skin color is among the most important qualitative traits on which selection is based in wine and table grape breeding programmes. Skin color is determined by the quantity and composition of anthocyanins. In prior work on cultivated forms, it was shown that polymorphisms in the grape transcription factor family VvMYBA are responsible for anthocyanin content variation in the berries of cultivated grapevine (Vitis vinifera subsp. vinifera). Wild grapevine (V. vinifera subsp. sylvestris) is the ancestor of the cultivated V. vinifera subsp. sativa and has black-colored berries. The purpose of this study was to determine how the VvmybA1 and VvmybA2 polymorphisms emerged and affected the genetic diversity of wild grapevines in the Mediterranean basin by examining samples from the Iberian Peninsula, Italian Peninsula and Caucasian region. Our observations provide evidence that variation in the two transcriptional regulators generated a novel allele series via length polymorphisms in VvmybA1 and a point mutation in VvmybA2, which is lacking in cultivated grapevine. Further, correlation was detected between allele composition and anthocyanin contents. According to polymorphisms in both VvMYBA genes at the color locus, we were able to identify several haplotypes. The most ancestral haplotype (HapN) was found in wild grapevine in the western Mediterranean region and corresponded to wine grape cultivars, whereas recent haplotypes were detected in eastern regions. These eastern zones showed the most diverse haplotypes, which appeared in table cultivars where intense breeding practices may have replaced the original haplotype diversity. These findings provide information about the evolution of grapes since their domestication and have direct implications for wine quality.

Study of intra-varietal diversity in biotypes of Aglianico and Muscat of Alexandria (Vitis vinifera L.) cultivars

Background and Aims Somatic mutations are the major source of intra-varietal variability in some species such as grapevine (Vitis vinifera L.) that are propagated vegetatively. Aglianico and Muscat of Alexandria are two cultivars mostly cultivated in the south of Italy, showing great intra-varietal diversity. The variability of 15 clones of Aglianico belonging to three biotypes, Taburno, Taurasi and Vulture, and 21 clones of Muscat of Alexandria, coming from France, Greece, Italy and Spain, was investigated. Methods and Results The intra-varietal diversity was investigated by ampelographic and eno-carpological (berry characteristics of oenological interest) methods (nine OIV descriptors, related to bunch and berry morphology, sugar content, total acid content and pH of must), chemical (anthocyanins, aromatic compounds and phenolic substances) and genetic analysis [18 K SNPs, single nucleotide polymorphisms (SNP)]. Intra-varietal diversity has been detected in both cultivars, mainly based on differences in the accumulation of secondary metabolites. The Taurasi biotype was discriminated by linear discrimination analysis mainly for the proportion of malvidin-3-O-glucoside acetate and methoxylated anthocyanins, while the Spanish Muscat biotype was discriminated based on the proportion of flavanols and hydroxycinnamic acids. Conclusions Ampelography and the concentration of secondary metabolites have proved to be the best tools for the characterisation of the intra-varietal variability of grapevine germplasm. The high-throughput genetic analysis based on 18 K SNP loci did not highlight a polymorphism sufficient to distinguish among biotypes of the same cultivar, suggesting that a different molecular approach has to be adopted. Significance of the Study This study confirms that intra-varietal diversity is better examined with a multidisciplinary approach.

Evidence for a Sympatric Origin of Ribolla gialla, Gouais Blanc and Schiava cultivars (V. vinifera L.)

Ribolla gialla is an autochthonous grape variety cultivated in Friuli Venezia Giulia (Italy) and in Slovenia, and probably originated in Eastern Central Europe. Just like Ribolla gialla, the Gouais blanc and Schiava cultivars also appear to have originated in the same geographical area. To verify this hypothesis, a pool of varieties probably sharing the same historical and geographical origin were taken into account. The fingerprinting, by 35 SSR loci, of Gouais blanc, Ribolla gialla, Schiava grossa and Schiava lombarda (synonyms of Schiava bresciana), is presented and the correlation among historical, geographical and genetic information of these cultivars was investigated. Gouais blanc and Traminer, already suggested as key varieties in the development of European grape diversity and as parents for some French varieties, interestingly enough seems to be linked to Ribolla gialla. The putative parentage was verified using 58 microsatellite markers. The genetic results suggest a common geographical origin for Gouais blanc, Ribolla gialla and the Schiava group: these cultivars appear to be related through sympatric origin. The hypothesis of Ribolla gialla as a progeny of Gouais blanc and Traminer was ruled out. The data proved a second-degree relationship between Gouais blanc and Ribolla gialla and a third or more distant degree of relationships between Ribolla gialla and Traminer.

Single nucleotide polymorphism profiles reveal an admixture genetic structure of grapevine germplasm from Calabria, Italy, uncovering its key role for the diversification of cultivars in the Mediterranean Basin

Background and Aims Grapevine (Vitis vinifera L.) is one of the earliest domesticated crops, dating back 6000 years ago in the Near East before spreading into Europe. Despite the abundance of historical, archaeological and genetic records, until now secondary domestication events in European regions are not well demonstrated. Here, a genetic characterisation of grape germplasm from Calabria in Southern Italy, a crucial area of the Mediterranean Basin, aims to validate this area as a secondary centre of crop domestication. Methods and Results True‐to‐type cultivar classification of 72 accessions was carried out by using microsatellite loci (simple sequence repeat) along with the main Organisation Internationale de la Vigne et du Vin ampelographic descriptors. The classification highlighted a high level of genetic diversity (He = 0.83) among native cultivars from Calabria. A decay of genetic diversity moving from southern Italy to north‐western Mediterranean regions was observed, probably because of repeated founder effects during the grapevine expansion from the Mediterranean Basin to Europe. Single nucleotide polymorphism analysis showed an admixture genetic structure at K = 7, clarifying a complex network of pedigree relationships generated by crosses among cultivars. Parentage analysis underlined a high proportion of parent–offspring relationships (76%) in Mantonico Bianco and Pecorello, hypothesising their key role in the pedigree of many native cultivars from the southern Mediterranean area. Conclusion Overall, our results appear to indicate a pivotal role of cultivars from Calabria in the grape genetic diversity of southern Italy. Furthermore, genetic analysis of grape wild accessions from Calabria should be of value in a discussion of a secondary centre of grape domestication. Significance of the Study A large grape collection from Calabria was for the first time characterised through ampelographic and genetic analysis.

Analysis of Genetic Structure of Twelve Sicilian Grapevine Cultivars

In recent years, Sicilian viticulture has shown a significant growth with a remarkable increase both in quantity and in quality. Sicily is a large wine region in Italy and its wines are already known on the international markets. Several autoch-thonous grapevine cultivars have been preserved in Sicily, but the Island genetic resources are still almost unexploited due to the lack of genetic characterization. The aim of the present study is to investigate molecular characteristics of Sicilian cultivars; evaluate probable cases of synonymies and false attributions, and preserve Sicilian germplasm. A wide screening of 12 major Sicilian cultivars was carried out (?Carricante?, ?Catarratto comune? and ?lucido?, ?Nero d?Avola?, ?Nerello cappuccio?, ?Nerello Mascalese?, ?Perricone?, ?Grillo?, ?Grecanico?, ?Inzolia?, ?Corinto nero? and ?Frappato?). In this frame a total of 687 accessions were collected and analysed with SSR markers (6 to 21 SSR loci). Allelic profiles were obtained and compared with international databases, allowing verification of mistakes and false attributions. Results clarified several synonymies and false attributions, and highlighted the richness and variability of the Sicilian grapevine platform still present in the region. Furthermore, these data provide a base for future studies.

Proteomic analysis among different aglianico ecotypes

Although molecular markers such as SSRs are extensively used for the identification of different vines, they often fail to discriminate the clones of a given cultivar. For these reasons, many attempts are made to find devices able to achieve this purpose. In this view, we evaluated the potential of proteomics in this field comparing the skin 2-DE profiles of berries belonging to three clones of the Italian cultivar ?Aglianico? (Taurasi, Taburno and Vulture) grown in the same field and considered in different moments to reduce the impact of environmental and seasonal variability. After a preliminary and completely automated gel alignment, we tested some multivariate statistical techniques to distinguish the three clones and to isolate the variables (i.e., the spots on the gels) responsible for the clonal discrimination. Although further analyses will be necessary to refine such an approach, the results are encouraging in order to identify markers useful in discriminating clones.

Italian wild grapevine : a state of the art on germplasm and conservation in 2010 : the Year of Biodiversity

Vitis vinifera subsp. sylvestris Gmel. is the wild progenitor of cultivated grapevine. In some countries, wild grapevines are considered on the brink of extinction. In 2006 the Italian Ministry of Environment funded a research project aiming to create a germplasm collection of wild grapevines pooled from existing Italian populations. In a two-year period, woody cuttings of 748 plants were collected and 246 among these are already in collection at the Cinque Terre National Park in Liguria, whose territory was designated a UNESCO world heritage site for its landscape agriculture and for the need to preserve its cultural, environmental, local and historical values. Samples collected have been genotyped at 20 SSR loci both to have a correct fingerprint of all the accessions and to check for possible mistakes during samplings. The ex situ collection is available for researchers interested in the study of the genetic structure of wild grapevine, both for population genetics and for studies regarding gene diversity.