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…

Modeling island and coastal vineyards potential in the context of climate change

Climate change impacts regional and local climates, which in turn affects the world’s wine regions. In the short term, these modifications rises issues about maintaining quality and style of wine, and in a longer term about the suitability of grape varieties and the sustainability of traditional wine regions. Thus, adaptation to climate change represents a major challenge for viticulture. In this context, island and coastal vineyards could become coveted areas due to their specific climatic conditions. In regions subject to warming, the proximity of the sea can moderate extremes temperatures, which could be an advantage for wine. However, coastal and island areas are particular prized spaces and subject to multiple pressures that make the establishment or extension of viticulture complex.
In this perspective, it seems relevant to assess the potentialities of coastal and island areas for viticulture. This contribution will present a spatial optimization model that tends to characterize most suitable agroclimatic patterns in historical or emerging vineyards according to different scenarios. Thanks to an in-depth bibliography a global inventory of coastal and insular vineyards on a worldwide scale has been realized. Relevant criteria have been identified to describe the specificities of these vineyards. They are used as input data in the optimization process, which will optimize some objectives and spatial aspects. According to a predefined scenario, the objectives are set in three main categories associated with climatic characteristics, vineyards characteristics and management strategies. At the end of this optimization process, a series of maps presents the different spatial configurations that maximize the scenario objectives.

1H-NMR-based Metabolomics to assess the impact of soil type on the chemical composition of Mediterranean red wines

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.

δ13C : A still underused indicator in precision viticulture  

The first demonstration of the interest of carbon isotope composition of sugars in grapevine, as an integrated indicator of vineyard water status, dates back to 2000 (Gaudillère et al., 1999; Van Leeuwen et al., 2001). Thanks to the isotopic discrimination of Carbon that takes place during plant photosynthesis, under hydric stress conditions, it is possible to accurately estimate the photosynthetic activity. Ever since, δ13C has been widely applied with success to zonation, terroir studies and vine physiology research, but is still not widely used by viticulturists. This is quite astonishing by considering the impact of global warming on viticulture and the need to improve water management, that would justify a widespread use of δ13C.
The lack of private laboratories proposing the analysis, the cost of the technology, as well as the long analytical delays, have been detrimental to its development. Some laboratories tried to overcome the analytical difficulties of isotopic analysis by using fourier transformed infrared spectroscopy, as a fast and cheap alternative to the official OIV method (IRMS). These claimed FTIR models have never been published or peer reviewed and cannot be considered robust. In this work, thanks to the recent acquisition of IRMS technology, new modern and robust applications of δ13C for viticulture are proposed. This includes the use of the analysis to make parcel separations at harvesting, the possibility to increase the precision of hydric stress cartography and the potential cost reduction when compared with Scholander pressure bomb analysis.

Soil quality in Beaujolais vineyard. Importance of pedology and cultural practices

A pedological study was carried out from 2009 to 2017 in Beaujolais vineyard, to improve physical and chemical knowledge of soils. It was completed in 2016 and 2017 by the current study, dealing with microbial aspects, in order to build a reference frame for improved advice in soil management. Microbial biomass was measured on representative plots of the six most common soil types identified in Beaujolais and, for each soil type, on plots with different levels of the main impacting parameters: total organic carbon, pH, cation exchange capacity, extractable copper. A total of 59 soil samples were collected. Confirming the results of various trials carried out in Beaujolais over the past 20 years, the results of the present study showed that the soils were still alive, but exhibited a large variability of biological parameters, which appeared dependant on both pedological and anthropic factors. Therefore, a good interpretation of biological parameters and advice for vine growers must rely on a pedologically-based referential with differentiated main driving factors. For example, the control of pH is of primary importance in granitic soils and in no way organic matter addition can improve soil quality if pH is too low. Conversely, in calcareous soils, biological parameters are more directly affected by direct or indirect (cover crops for example) inputs of organic matter. The use of biological parameters, such as microbial biomass, is of great potential value to improve advice on agro-viticultural practices (soil management, fertilization, liming, etc.), basis of a sustainable wine production on fragile soils.

A predictive model of spatial Eca variability in the vineyard to support the monitoring of plant status

[lwp_divi_breadcrumbs home_text="IVES" use_before_icon="on" before_icon="||divi||400" module_id="publication-ariane" _builder_version="4.19.4" _module_preset="default" module_text_align="center" module_font_size="16px" text_orientation="center"...