Macrowine 2021
IVES 9 IVES Conference Series 9 Capture depletion of grapevine DNA: an approach to advance the study of microbial community in wine

Capture depletion of grapevine DNA: an approach to advance the study of microbial community in wine

Abstract

The use of next-generation sequencing (NGS) has helped understand microbial genetics in oenology. Current studies mainly focus on barcoded amplicon NGS but not shotgun sequencing, which is useful for functional analyses. Since the high percentage of grapevine DNA conceals the microbial DNA in must, the majority of sequencing data is wasted in bioinformatic analyses. Here we present capture depletion of grapevine whole genome DNA. By using biotinylated RNA baits made from genomic grapevine libraries, the grapevine DNA is hybridized to the baits and captured by magnetic beads while the targeted microbial DNA is left and subsequently sequenced. A pilot study was carried out to mimic degraded complex materials. Using known complex microbial samples and spiking in large portion of grapevine DNA, the hybridization and depletion took place with quantification by qPCR. Different parameters, such as hybridization hours, temperatures and bait amounts were also tested for potential effects. The aim is to provide an insight into depletion of undesired grapevine DNA and to enhance the data yield in NGS studies of microbial DNA with a more cost effective way suitable for future oenological studies.

Publication date: May 17, 2024

Issue: Macrowine 2016

Type: Poster

Authors

Sarah S.T. Mak*, Anders Hansen, Christian Carøe, Jacob Enk, Jean-Marie Rouillard, M Thomas P Gilbert

*Centre for GeoGenetics

Contact the author

Tags

IVES Conference Series | Macrowine | Macrowine 2016

Citation

Related articles…

Effects of post-fermentative cold maceration on chemical and sensory characteristics of Syrah, Cabernet Franc and Montepulciano wines

Astringency sensation decreases slowly during the aging of red wine. Complex reactions of condensation and precipitation of wine polyphenols are involved in this phenomenon. Wine composition and conditions of aging, such as temperature and oxygen availability, strongly influence evolution of the phenol matrix. Recently, a Post-Fermentative cold Maceration (PFM) technique was tested with the aim of accelerating reactions leading to the reduction of astringency and exploiting chemical compounds not extracted from the solid parts of grapes during the previous traditional maceration phase. To this purpose, an innovative maceration system was engineered and used to perform PFM trials on marc derived from vinification of different varieties of red grapes.

Phenolic profiles of minor red grape cultivars autochthonous from the Spanish region of La Mancha

The phenolic profiles of little known red grape cultivars, namely Garnacho, Moribel and Tinto Fragoso, which are autochthonous from the Spanish region of La Mancha (ca. 600,000 ha of vineyards) have been studied over the consecutive seasons of years 2013 and 2014. The study was separately performed over the skins, the pulp and the seeds, and comprised the following phenolic types: anthocyanins, flavonols, hydroxycinnamic acid derivatives (HCADs), total proanthocyanidins (PAs) and their structural features. The selected grape cultivars belong to the Vine Germplasm Bank created in this region in order to preserve the great diversity of genotypes grown in La Mancha.

Some applications come from a method to concentrate proteins

All techniques usually used to assay proteins was not reliable in vegetable extract due to interferences with the components included in extracts like polyphenols, tanins, pectines, aromatics compounds. Absorbance at 280nm, Kjeldhal assay, Biuret and Lowry methods, Acid Bicinchonique technique and Bradford assay give the results depending on the composition of extract, on the presence or not of detergent and on the raw material (Marchal, 1995). Another difficulty in these extracts for the quantification of proteins comes from the large amount of water included in vegetable and the low concentration of proteins. Thus in red wines, proteins are usually not taken into account due to their low concentration (typically below 10 mgL-1) and to the presence of anthocyanis and polyphenols.

Novel contribution to the study of mouth-feel properties in wines

In general, there is a well-established lexicon related to wine aroma and taste properties; however mouth-feel-related vocabulary usually includes heterogeneous, multimodal and personalized terms. Gawel et al.
(2000) published a wheel related to mouthfeel properties of red wine. However, its use in scientific publications has been limited. The authors accepted that the approach had certain limitations as it included redundant and terms with hedonic tone and some others were absent. It is of high interest to generate a mouth-feel lexicon and finding the chemical compound or group of compounds responsible for such properties in red wine. In the present work a chemical fractionation method has been developed.

Dissecting the polysaccharide‐rich grape cell wall matrix during the red winemaking process, using high‐throughput and fractionation methods

Limited information is available on grape wall-derived polymeric structure/composition and how this changes during fermentation. Commercial winemaking operations use enzymes that target the polysaccharide-rich polymers of the cell walls of grape tissues to clarify musts and extract pigments during the fermentations. In this study we have assessed changes in polysaccharide composition/ turnover throughout the winemaking process by applying recently developed cell wall profiling approaches to both wine and pomace polysaccharides. The methods included gas chromatography for monosaccharide composition (GC-MS), infra-red (IR) spectroscopy and comprehensive microarray polymer profiling
(CoMPP) using cell wall probes.