A new vector, pGID052, for genetic transfer in<i>Oenococcus oeni</i>

Beltramo, Charlotte; Oraby, Mona M.; Bourel, GÃ©rald; Garmyn, Dominique; Guzzo, Jean

doi:10.1111/j.1574-6968.2004.tb09626.x

Cited by 14 publications

(16 citation statements)

References 34 publications

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“…However, the constitution of biodiversity pool remains a very important step in the case of O. oeni , as effective DNA recombinant tools have been only recently set up for this species (Zuniga et al. 2003; Beltramo et al. 2004; Assad‐García et al.…”

Section: Discussionmentioning

confidence: 99%

Characterization and technological properties of Oenococcus oeni strains from wine spontaneous malolactic fermentations: a framework for selection of new starter cultures

Solieri

Genova²,

Paola³

et al. 2010

Journal of Applied Microbiology

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Aims: To characterize the genetic and phenotypic diversity of 135 lactic acid bacteria (LAB) strains isolated from Italian wines that undergone spontaneous malolactic fermentation (MLF) and propose a multiphasic selection of new Oenococcus oeni malolactic starters. Methods and Results: One hundred and thirty‐five LAB strains were isolated from 12 different wines. On the basis of 16S amplified ribosomal DNA restriction analysis (ARDRA) with three restriction enzymes and 16S rRNA gene sequencing, 120 O. oeni strains were identified. M13‐based RAPD analysis was employed to investigate the molecular diversity of O. oeni population. Technological properties of different O. oeni genotypes were evaluated in synthetic medium at increasing selective pressure, such as low pH (3·5, 3·2 and 3·0) and high ethanol values (10, 11 and 13% v/v). Finally, the malolactic activity of one selected strain was assessed in wine by malolactic trial in winery. Conclusions: The research explores the genomic diversity of wine bacteria in Italian wines and characterizes their malolactic metabolism, providing an efficient strategy to select O. oeni strains with desirable malolactic performances and able to survive in conditions simulating the harsh wine environment. Significance and Impact of the Study: This article contributes to a better understanding of microbial diversity of O. oeni population in Italian wines and reports a framework to select new potentially O. oeni starters from Italian wines during MLF.

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Section: Discussionmentioning

confidence: 99%

Characterization and technological properties of Oenococcus oeni strains from wine spontaneous malolactic fermentations: a framework for selection of new starter cultures

Solieri

Genova²,

Paola³

et al. 2010

Journal of Applied Microbiology

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“…These studies will provide an essential contribution to the understanding of the cell physiology of O. oeni. In order to carry out gene inactivation in O. oeni, future studies will focus on the construction of an integrative vector for gene disruption derived from the conjugative plasmid pGID052 (5).…”

Section: Discussionmentioning

confidence: 99%

CtsR Is the Master Regulator of Stress Response Gene Expression inOenococcus oeni

et al. 2005

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“…Complementary oligonucleotides psynC and psynD (50 mM each) ( Table 2) were annealed by heating (95°C, 5 min) and gently cooling (95°C to 20°C), and the DNA duplex was restricted with SwaI and SmaI restriction enzymes and then subcloned between the SwaI and PvuII sites of plasmid pGID052 (32). The resulting plasmid, named pSYNS3 (Table 1), was then used as a DNA matrix for PCR amplification of the promoter region using oligonucleotides olcg302 and olcg305 (Table 2).…”

Section: Methodsmentioning

confidence: 99%

The Antisense RNA Approach: a New Application for In Vivo Investigation of the Stress Response of Oenococcus oeni, a Wine-Associated Lactic Acid Bacterium

Darsonval

Msadek

Alexandre

et al. 2016

Appl Environ Microbiol

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Oenococcus oeni is a wine-associated lactic acid bacterium mostly responsible for malolactic fermentation in wine. In wine, O. oeni grows in an environment hostile to bacterial growth (low pH, low temperature, and ethanol) that induces stress response mechanisms. To survive, O. oeni is known to set up transitional stress response mechanisms through the synthesis of heat stress proteins (HSPs) encoded by the hsp genes, notably a unique small HSP named Lo18. Despite the availability of the genome sequence, characterization of O. oeni genes is limited, and little is known about the in vivo role of Lo18. Due to the lack of genetic tools for O. oeni, an efficient expression vector in O. oeni is still lacking, and deletion or inactivation of the hsp18 gene is not presently practicable. As an alternative approach, with the goal of understanding the biological function of the O. oeni hsp18 gene in vivo, we have developed an expression vector to produce antisense RNA targeting of hsp18 mRNA. Recombinant strains were exposed to multiple stresses inducing hsp18 gene expression: heat shock and acid shock. We showed that antisense attenuation of hsp18 affects O. oeni survival under stress conditions. These results confirm the involvement of Lo18 in heat and acid tolerance of O. oeni. Results of anisotropy experiments also confirm a membrane-protective role for Lo18, as previous observations had already suggested. This study describes a new, efficient tool to demonstrate the use of antisense technology for modulating gene expression in O. oeni. Oenococcus oeni is a wine-associated lactic acid bacterium mostly responsible for wine malolactic fermentation (MLF) (1). In wine medium, O. oeni evolves under extreme physicochemical conditions: low pH (3.2 to 3.5), low temperature (18°C), and the presence of biological competitors such as yeast, producing inhibitory compounds, i.e., fatty acids, ethanol, 2-phenylethanol, and sulfites. The winemaking process itself may also constitute a hostile factor for bacterial growth. All these hostile environmental factors can cause a delay in MLF initiation. Thus, this fermentation step is currently not yet fully managed. However, the gradual increase in the ethanol level in wine during the alcoholic fermentation performed by yeasts and the production of substances inhibiting growth of lactic acid bacteria (dodecanoic acid or decanoic acid) led to the selection of the species most suited and adapted to wine conditions: O. oeni (1, 2). A better understanding of the mechanisms involved in stress-adaptive responses is essential to improve O. oeni development in wine and to perfect industrial processes of preparing wine malolactic starter. Because of its acidophilic properties, O. oeni is an interesting model for investigating stress response mechanisms in lactic acid bacteria (LAB). Like any organism undergoing an environmental stress, O. oeni tries to restore a metabolic profile beneficial for survival. Over the past decades, several mechanisms involved in adaptation to wine were described, inclu...

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A new vector, pGID052, for genetic transfer inOenococcus oeni

Cited by 14 publications

References 34 publications

Characterization and technological properties of Oenococcus oeni strains from wine spontaneous malolactic fermentations: a framework for selection of new starter cultures

Characterization and technological properties of Oenococcus oeni strains from wine spontaneous malolactic fermentations: a framework for selection of new starter cultures

CtsR Is the Master Regulator of Stress Response Gene Expression inOenococcus oeni

The Antisense RNA Approach: a New Application for In Vivo Investigation of the Stress Response of Oenococcus oeni, a Wine-Associated Lactic Acid Bacterium

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