Evaluation of intra-specific diversities in Oenococcus oeni through analysis of genomic and expressed DNA.
In winemaking Oenococcus (O.) oeni is the most frequent species of lactic acid bacteria (LAB) associated with malolactic fermentation (MLF). Several studies have demonstrated that O. oeni is a quite homogeneous species and strains are difficult to differentiate especially when isolates from the same region are analyzed. In this study, the molecular biodiversity of O. oeni isolated from wines of the same region (Aglianico produced in Basilicata Region, Southern Italy) was evaluated with the aim of designing a molecular approach for discrimination and characterization of the isolates at the strain level. Three molecular techniques were applied: random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), restriction endonucleases analysis-pulsed field gel electrophoresis (REA-PFGE) and differential display PCR (DD-PCR). The results obtained by RAPD-PCR confirmed the difficulty in differentiating isolates. By means of REA-PFGE a higher polymorphism, often related to the origin (winery) of strains, was revealed. However, on analyzing strains isolated from the same winery, only in some cases was more than one REA-PFGE pattern obtained. By analyzing dendrograms constructed on the basis of DD-PCR profiles differentiation of strains isolated from the same winery, in some cases, could be accomplished. The reliability of the DD-PCR in the differentiation of closely related strains suggests that this method could represent an alternative and/or additional tool to other molecular methods, such as REA-PFGE, for fine characterization of oenococcal strains.
Lechiancole
Tiziana
T
Dipartimento di Biologia, Difesa e Biotecnologie Agro-Forestali, Università degli Studi della Basilicata, Campus Macchia Romana 100 Potenza, Italy.
Blaiotta
Giuseppe
G
Messina
Dina
D
Fusco
Vincenzina
V
Villani
Francesco
F
Salzano
Giovanni
G
D004269:DNA, Bacterial / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D005285:Fermentation; D016680:Genome, Bacterial / Q000235:genetics; D006095:Gram-Positive Cocci / Q000145:classification* / Q000235:genetics*; D008291:Malate Dehydrogenase / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D015203:Reproducibility of Results; D014920:Wine / Q000382:microbiology
D004269:DNA, Bacterial; C037731:malolactic enzyme; D008291:Malate Dehydrogenase; D015252:Deoxyribonucleases, Type II Site-Specific; C038485:GGGCCC-specific type II deoxyribonucleases