OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Grape and wine microorganisms: diversity and adaptation 9 Use of antisense RNA technology to modulate gene expression in Œnococcus oeni

Use of antisense RNA technology to modulate gene expression in Œnococcus oeni

Abstract

Œnococcus oeni is a wine-associated lactic acid bacterium performs the malolactic fermentation, which improves the taste and aromatic complexity of many wine. Although, wine exhibits harsh and challenging conditions (low pH, low temperature, nutrient-poor and presence of ethanol), O. oeni possesses a remarkable adaptability to those physiochemical conditions.

Mechanisms for responding to environmental changes are universally present in living beings and are essential for coping with the stress and for adapting to the new conditions. O. oeni tolerance to low pH and ethanol make its an interesting bacteria model for investigating stress response mechanism in lactic acid bacteria. However, lack of appropriate techniques to manipulate O. oeni genome has long delay molecular study of this fastidious bacterium. To get around the lack of genetic tool for gene replacement, we focused our work on gene inactivation by using antisense RNA approach to modulate gene expression. With the goal to understanding the function of O. oeni hsp genes in vivo, we produce antisense RNA targeting genes encoding: a small Hsp (hsp18)1, the master regulator of stress response (ctsR)2 and two caseinolytic protease L members of the HSP100 chaperone family (clpL1, clpL2). Thereby, we highlighted that in vivo inhibition of the expression of some of these genes strongly affects the survival of O. oeni in stress conditions.

This study presents an elegant approach providing access to an in vivo study of gene function in O. oeni.

References:

1. Darsonval, M., Msadek, T., Alexandre, H. & Grandvalet, C. The Antisense RNA Approach: a New Application for In Vivo Investigation of the Stress Response of Oenococcus oeni, a Wine-Associated Lactic Acid Bacterium. Appl. Environ. Microbiol. 82, 18–26 (2016).

2. Darsonval, M., Julliat, F., Msadek, T., Alexandre, H. & Grandvalet, C. CtsR, the Master Regulator of Stress-Response in Oenococcus oeni, Is a Heat Sensor Interacting With ClpL1. Front Microbiol 9, (2018).

DOI:

Publication date: June 10, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Cosette Grandvalet, Frédérique Julliat, Maud Darsonval, Tarek Msadek, Hervé Alexandre

UMR A 02.102 Procédés Alimentaires et Microbiologiques, AgroSup Dijon – Université de Bourgogne Franche-Comté, Dijon, FRANCE.
Unité de Biologie des Bactéries Pathogènes àGram Positif, Institut Pasteur, Paris, FRANCE.
CNRS ERL 6002, Paris, FRANCE.

Contact the author

Keywords

Oenococcus oeni, lactic acid bacteria, antisense RNA , stress response 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

Chitosan treatment to manage grapevine downy mildew

Downy mildew is one of the most important grapevine diseases, caused by the Oomycete Plasmopara viticola. The management of the disease in organic agriculture can require up to 15 copper applications per year. However, copper accumulates in the soil, is phytotoxic and is toxic for organisms living in the soil, its use has been restricted in European Union to maximum 28 kg in 7 years. Therefore, testing of alternatives with equal effectiveness is desirable. Among those, the natural biopolymer chitosan, obtained from crab shells, proved to be effective toward downy mildew in plot experiments. The aim of our trials was to extend chitosan applications in large scale experiments in different years, cultivars and environmental conditions.

Hierarchy of the interactions between physical and biological parameters intervening in the Pyrenean Gascon foothill vineyard

Un travail sur les A.O.C. du piémont pyrénéen occidental permet de construire une hiérarchie de paramètres climatiques, géo-pédologiques, morphologiques, de saisir leurs niveaux d’interaction et d’élaborer une méthodologie pour proposer un zonage

The true cost of the vineyard landscape enhancement. First results in the Venezia biodistrict 

The research is part of the “Ecovinegoals” project, financed by Interreg Adrion funds. It aims to encourage the adoption and dissemination of agroecological practices in intensive wine-growing areas. The study focuses on cost analysis of the wine-growing landscape enhancement in an organic winery in order to provide a useful tool for winemakers to direct their investments in green infrastructures. One of the Italian pilot areas of the Ecovinegoals project is the Venezia Biodistrict, characterized by viticulture in a flat reclamation area of 105,800 hectares.

Research on the origin and the side effects of chitosan stabilizing properties in wine

Fungal chitosan is a polysaccharide made up of glucosamine and N-acetyl-glucosamine and derived from chitin-glucan of Aspergillus niger or Agaricus bisporus. Fungal chitosan has been authorized as an antiseptic agent in wine since 2009 (OIV) and in organic wine in 2018. At the maximum dose of 10g/hl, it was shown to eliminate Brettanomyces bruxellensis, the main spoilage agent in red wines. Fungal chitosan is highly renewable, biocompatible (ADI equivalent to sucrose) and non-allergenic. However, winemakers often prefer to use sulfites (SO2), though sulfites are classified as priority food allergens, than chitosan. Indeed, many conflicting reports exist regarding its efficiency and its side effects towards beneficial wine microorganisms or wine taste. These contradictions could be explained by the heterogeneity of the fungal chitosan lots traded, the diversity of the wines (chemical composition, winemaking process), but also, by the recently highlighted huge genetic diversity prevailing in wine microbial species.

La zonazione della valle d’Illasi (Verona)

In the bottom of Val d’Illasi (Verona province), one of the major valleys which passes through the Lessini mountains, viticulture is widely extended. In the territory belonging to Illasi and Tregnago villages, which includes ca. 1100 ha of vineyards, devoted to produce Soave and Valpolicella DOC wines, an experimental survey was conducted on a network of twenty five reference vineyards.