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…

Using combinations of recombinant pectinases to elucidate the deconstruction of the polysaccharide‐rich grape cell wall during winemaking

The effectiveness of enzyme-mediated maceration processes in red winemaking relies on a clear picture of the target (berry cell wall structure) to achieve the optimum combination of specific enzymes to be used. However, we lack the information on both essential factors of the reaction (i.e. specific activities in commercial enzyme preparation and the cell wall structure of berry tissue). In this study, the different combinations of pure recombinant enzymes and the recently validated high throughput cell wall profiling tools were applied to extend our knowledge on the grape berry cell wall polymeric deconstruction during the winemaking following a combinatorial enzyme treatment design.

Characterizing the effects of nitrogen on grapevines with different scion/rootstock combinations: agronomic, metabolomic and transcriptomic approaches

Most vineyards are grafted and include a variety (Vitis vinifera) grafted over a wild Vitis rootstock (hybrids of V. berlandieri, riparia and rupestris). Grape berry quality at harvest depends on a subtle balance between acidity and the concentrations of sugars, polyphenols and precursors of aroma compounds. The mechanisms controlling the balance of sugars/acids/polyphenols are influenced by the abiotic environment, in particular nitrogen supply, and interact with the genotypes of both the scion variety and the rootstock. Previous work suggests that some of the effects of water stress are in fact linked to a nitrogen deficiency driven indirectly by the reduction of water absorption.

The moment of preharvest elicitor application influence its final effect on winegrapes quality

Phenolic compounds are secondary metabolites of grapes. Plants produce a wide variety of this type of metabolites through diverse biosynthesis pathways and their production is sometimes a response to external stimuli, either environmental or biotic stresses. Some of them may act as chemical defenses against pathogens or herbivores and their synthesis is increased when the attack exists. However, it is remarkable that the synthesis of these interesting compounds can be activated even when the stimulus is not present, with the use of elicitors. These are substances that when applied exogenously trigger the biosynthetic pathways conducting to the synthesis of these defense compounds.

Sensory definition of green aroma concept in red French wines. Evidence for the contribution of novel volatile markers

The aromatic complexity of a wine results from the perception of the association of volatile molecules and each aroma can be categorized into different families. The “green” aromas family in red wines has retained our attention by its close link with the fruity perception. In that study, the “green” olfactory concept of red wines was considered through a strategy combining both sensory analysis and hyphenated chromatographic techniques including HPLC and MDGC (Multidimensional Gas Chromatography). The aromatic space of this concept was specified by lexical generation through a free association task on 22 selected wines by a panel of wine experts. Then, 70 French red wines were scored on the basis of the intensity of their “green” and “fruity” attributes.

Analysis of voltammetric fingerprints of different white grape musts reveals genotype-related oxidation patterns

Must oxidation is a complex process involving multiple enzymatic transformations, including the oxidation of phenolics containing an ortho-diphenol function. The latter process has a primary influence on wine aroma characteristics and stability, due to the central role of ortho-diphenols in the non-enzymatic oxidative reactions taking place during winemaking and in finished wine. Although oxidation of must is traditionally avoided, in recent years its contribution to wine quality has been revisited, and in some cases improvements to wine aroma have been observed with the application of controlled must oxidation. Nowadays there is a great interest in the wine industry towards the identification of specific markers or patterns to characterize and classify the response of grape must to oxidation.