title: Dynamics of lactic acid bacteria populations in Rioja wines by PCR-DGGE, comparison with culture-dependent methods.
abstract: Lactic acid bacteria populations of red wine samples from industrial fermentations, including two different vinification methods were studied. For this investigation, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis was employed to supplement previous results that were obtained by culture-dependent methods. PCR-DGGE was aimed to study two targeted genes, 16S ribosomal DNA (rDNA) and rpoB, and the results were useful to evaluate the microbial populations in wine samples. Moreover, an improvement of a detection limit determined so far for DGGE analysis was obtained with the method described in this study, what made possible to identify lactic acid bacteria populations below 10(1) colony-forming unit/mL. The species Oenococcus oeni was the most frequently detected bacterium, but identifications close to species Oenococcus kitaharae and Lactococcus lactis that are not often found in wine were firstly identified in samples of this research. PCR-DGGE allowed to detect 9 out of 11 lactic acid bacteria species identified in this study (nine by PCR-16S rDNA/DGGE and four by PCR-rpoB/DGGE), while five species were detected using the modified de Man, Rogosa and Sharpe agar. Therefore, the two methods were demonstrated to be complementary. This finding suggests that analysis of the lactic acid bacteria population structure in wine should be carried out using both culture-dependent and culture-independent techniques with more than one primer pair.
authors.affiliation: Instituto de Ciencias de la Vid y del Vino (Gobierno de La Rioja, Universidad de La Rioja, CSIC), C/ Madre de Dios 51, 26006, Logroño, La Rioja, Spain.
authors.forename: Lucía
authors.initials: L
authors.lastname: González-Arenzana
authors.forename: Rosa
authors.initials: R
authors.lastname: López
authors.forename: Pilar
authors.initials: P
authors.lastname: Santamaría
authors.forename: Isabel
authors.initials: I
authors.lastname: López-Alfaro
mesh_terms: D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D017045:Lactococcus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D014920:Wine / Q000382:microbiology*
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S