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…

Metabolomics of grape polyphenols as a consequence of post-harvest drying: on-plant dehydration vs warehouse withering

A method of suspect screening analysis to study grape metabolomics, was developed [1]. By performing ultra-high performance liquid chromatography (UHPLC) – high-resolution mass spectrometry (HRMS) analysis of the grape extract, averaging 320-450 putative grape compounds are identified which include mainly polyphenols. Identification of metabolites is performed by a new HRMS-database of putative grape and wine compounds expressly constructed (GrapeMetabolomics) which currently includes around 1,100 entries.

On the losses of dissolved CO2 during champagne aging

A misconception lingers in the minds of some wine consumers that Champagne wines don’t age. It’s largely a myth, certainly as far as the best cuvees are concerned. Actually, during the so-called autolysis period of time (in the closed bottle, after the “prise de mousse”), complex chemical reactions take place when the wine remains in contact with the dead yeast cells, which progressively bring complex and very much sought-after aromas to champagne. Nevertheless, despite their remarkable impermeability to liquid and air, caps or natural cork stoppers used to cork the bottles are not 100% hermetic with regard to gas transfers. Gas species therefore very slowly diffuse through the cap or cork stopper, along their respective inverse partial pressure. After the “prise de mousse”, because the partial pressure of CO2 in the bottleneck reaches up to 6 bars (at 12 °C), gaseous CO2 progressively diffuse from the bottle to the ambient air
(where the partial pressure of gaseous CO2 is only of order of 0,0004 bar).

Pesticide removal in wine with a physical treatment by molecular sieving

All along the winemaking process, conditioning and aging, wine is susceptible to be contaminated by different molecules. Contaminations can have various origins, related to wine microorganisms or as a result of an exogenous contamination. The aforementioned contamination of the wine can be caused by the migration of molecules from the materials in contact with the wine or by a contamination from exogenous molecules present in the air. Regardless of the source of the contamination, mainly two types of consequences can be observed.

Use of chitosan as a secondary antioxidant in juices and wines

Chitosan is a polysaccharide produced from the deacetylation of chitin extracted from crustaceous and fungi. In winemaking chitosan is mainly used in the clarification of grape juice and wine, stabilization of white wines, removal of metals and to prevent wine spoilage by undesired microorganisms. The addition of chitosan to model wine systems was able to retard browning, reduce levels of metallic ions (Fe and Cu) and to protect varietal thiols due to its antiradical activity1. The present experiment was planned in order to evaluate the use of chitosan as a secondary antioxidant at three different stages of Sauvignon blanc fermentation and winemaking. Sauvignon blanc juices from three different locations were obtained at a commercial winery in Marlborough, New Zealand. One lots of grapes was collected from a receival bin and pressed into juice with a water-bag press, and a further juice sample was collected from a commercial pressing operation. Chitosan (1 g/L, low molecular weight, 75 – 85% deacetylated) was added to the juice after pressing, after cold settling, after fermentation, or at all these stages. Controls without any chitosan additions were also prepared.

Colour assessment of port wines using colorimetric and spectrophotometric methods

Colour is an important quality parameter in wines and is the result of a complex mixture of pigments
(including anthocyanins and their derivatives, quinones, xanthyllium compounds, etc.). Red wine colour changes over time as pigments react between themselves and with other wine macromolecules
(particularly polyphenols). During wine tasting, colour is normally assessed on the outer rim of the wine profile in a tilted glass, since most wines are too opaque to be analysed in the middle of the glass. Therefore, depending on the depth of observation considered, the perception of wine colour can be different.