In Vivo PCR-DGGE Analysis of <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> and <Emphasis Type="Italic">Oenococcus oeni</Emphasis> Populations in Red Wine |
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Authors: | G Spano A Lonvaud-Funel O Claisse S Massa |
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Institution: | (1) Department of Food Science, Foggia University, via Napoli 25, 71100 Foggia, Italy;(2) Faculté d’Oenologie UMR INRA, Laboratoire de Biotechnologie et Microbiologie Appliquée (LBMA) 351, cours de la libération, Talence cedex, France |
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Abstract: | In order to monitor Lactobacillus plantarum and Oenococcus oeni in red wine produced with Italian grape (variety “Primitivo di Puglia”), a polymerase chain reaction– denaturing gradient
gel electrophoresis (PCR-DGGE) approach using the rpoB as gene target was established. Wine was treated or not with potassium metabisulphite and supplemented with a commercial
bacterial starter of O. oeni to encourage malolactic fermentation. Samples were taken from the vinification tanks at 4, 10, 16, 22, and 28 days after
the start of alcoholic fermentation. Genomic DNA was directly isolated from wine and identification of lactic acid bacteria
was performed using primers rpoB1, rpoB1O, and rpoB2 able to amplify a region of 336 bp corresponding to the rpoB gene. Amplified fragments were separated in a 30–60% DGGE gradient, and the ability of the PCR-DGGE analysis to distinguish
L. plantarum and O. oeni was assessed. The results reported suggest that the PCR-DGGE method, based on the rpoB gene as molecular marker, is a reproducible and suitable tool and may be of great value for wine makers in order to monitor
spoilage microorganisms during wine fermentation. |
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