Recovery and purification of proteins from grape seed byproducts using proteomic and separative techniques

Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.

Copper is listed among the active substances candidates for substitution (Regulation EU 2015/408). Yet still, because of the lack of valid alternatives, the European Commission recently confirmed its usage authorization by limiting the maximum amount to 28 Kg per hectare in 7 years, i.e. an average of 4 kg/year (Reg. EU 2018/1981).This restriction is due to copper accumulation in soils and surface waters both caused by a steady application, especially on perennial crops (Riepert et al., 2013). The aim of this work is to determine if treatments with reduced copper dosages are able to reach different grapevine’s organs, with particular focus on the core of bunches, and if these small amounts can ensure the respect of the legislative prescription, without compromising the phytosanitary conditions of the vineyards, thus grape yields.

The use of pectinolytic enzymes in winemaking is state of the art. These enzymes catalyse the degradation of pectic substances through depolymerization (hydrolases and lyases) and de-esterification. As a result, it supports the extraction of juice and facilitates filtration. It has also been shown in winemaking that the presence of pectinolytic enzymes improves the stability, taste, texture, colour and aroma of products. With regard to enzymes currently applied in winemaking, enzymes derived from filamentous fungi dominate the enzyme industry. Fungal-based pectinolytic enzymes specifically require purification from the culture medium to eliminate unwanted side reactions, which is poorly sustainable. Some non-traditional yeast strains have been reported to exhibit pectinolytic activities. Therefore, the direct use of pectinolytic yeast during wine fermentation process can be an attractive and alternative source for the use of enzymes as input.

Among the Vitis vinifera L. cv. Moscato, Moscato Bianco is the oldest and most cultivated one in Europe (1). According to the OIV Focus 2015, Italy is the country with the largest cultivated area of Moscato Bianco with about 12500 hectares (2), that is used to produce well-known wines (i.e., Moscato Passito in Piedmont, Moscato di Trani in Puglia, and Moscatello di Montalcino in Tuscany), mainly obtained from partially dehydrated grapes (1). Different dehydration techniques can strongly modify the chemical compounds of oenological interest, among which Volatile Organic Compounds (VOCs) (1) that are the main responsible for the varietal sensory character of the final wine.

Disease resistant hybrid grape cultivars are now allowed in a number of EU wine PDOs, and are also accepted in a number of countries outside the EU. There is increasing interest in diseases resistant hybrid grape cultivars (RHGCs) because they allow for the production of healthy, high quality grapes with limited use of pesticides and the associated environmental and public health