Manna Crespan

Direct multiplex PCR for grapevine genotyping and varietal identification

Grapevine cultivar identification is based mainly on two complementary methodologies: microsatellite (simple sequence repeat (SSR)) DNA analysis and traditional ampelography. Here, we report a direct multiplex PCR approach that allows the simultaneous amplification of 11 SSR loci from crude samples, i.e. bypassing DNA extraction, by using an engineered DNA polymerase improved to tolerate plant PCR inhibitors. Many different plant tissues were successfully amplified: leaf, root, wood, berry flesh and skin, stalk and must, from wine and table grape varieties, and rootstocks. The direct multiplex PCR that we propose is quicker and cheaper than the methodologies used until now, and provides accurate results. Thus, SSR DNA analysis becomes economically more accessible to a larger number of potential users in addition to research institutes.

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

The Geographic Distribution of Saccharomyces cerevisiae Isolates within three Italian Neighboring Winemaking Regions Reveals Strong Differences in Yeast Abundance, Genetic Diversity and Industrial Strain Dissemination

In recent years the interest for natural fermentations has been re-evaluated in terms of increasing the wine terroir and managing more sustainable winemaking practices. Therefore, the level of yeast genetic variability and the abundance of Saccharomyces cerevisiae native populations in vineyard are becoming more and more crucial at both ecological and technological level. Among the factors that can influence the strain diversity, the commercial starter release that accidentally occur in the environment around the winery, has to be considered. In this study we led a wide scale investigation of S. cerevisiae genetic diversity and population structure in the vineyards of three neighboring winemaking regions of Protected Appellation of Origin, in North-East of Italy. Combining mtDNA RFLP and microsatellite markers analyses we evaluated 634 grape samples collected over 3 years. We could detect major differences in the presence of S. cerevisiae yeasts, according to the winemaking region. The population structures revealed specificities of yeast microbiota at vineyard scale, with a relative Appellation of Origin area homogeneity, and transition zones suggesting a geographic differentiation. Surprisingly, we found a widespread industrial yeast dissemination that was very high in the areas where the native yeast abundance was low. Although geographical distance is a key element involved in strain distribution, the high presence of industrial strains in vineyard reduced the differences between populations. This finding indicates that industrial yeast diffusion it is a real emergency and their presence strongly interferes with the natural yeast microbiota.

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

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.