16S-ARDRA, a tool for identification of lactic acid bacteria isolated from grape must and wine.
Lactic acid bacteria (LAB) are found in a great variety of habitats, including grape must and wines. There is a close relationship between the species of LAB which develop during fermentation and the eventual quality of the wine. For these reasons analytical techniques allowing fast and reliable identification of wine LAB are needed. In this work a simple and accurate protocol for identifying species of LAB isolated from grape must and wine is presented. This protocol is based on the amplification, directly from colony, of 16S rDNA and later digestion with one of the following restriction enzymes BfaI, MseI and AluI. A sequential use of the three enzymes is proposed to simplify LAB wine identification, first MseI, then BfaI and finally, if necessary, AluI digestion. The technique was able to discriminate 32 of the 36 LAB reference species tested and allowed the identification of 342 isolates from musts and wines. The isolates belonged to the species: Lactobacillus brevis, L. collinoides, L. coryniformis, L. bilgardii, L. mali, L. paracasei, Leuconostoc mesenteroides, Oenococcus oeni, Pediococcus parvulus and P. pentosaceus.
Rodas
Ana María
AM
ENOLAB--Laboratori de Microbiologia Enològica, Departament de Microbiologia i Ecologia, Facultat de Ciències Biològiques, Universitat de València, Burjassot, València, Spain.
Ferrer
Sergi
S
Pardo
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D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004275:DNA, Ribosomal / Q000032:analysis* / Q000235:genetics; D005285:Fermentation; D005516:Food Microbiology; D006094:Gram-Positive Bacteria / Q000145:classification* / Q000302:isolation & purification* / Q000378:metabolism; D006601:Hexoses / Q000378:metabolism; D019344:Lactic Acid / Q000096:biosynthesis; D007778:Lactobacillus / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D007934:Leuconostoc / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D010376:Pediococcus / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D014920:Wine / Q000382:microbiology*
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D006601:Hexoses; D019344:Lactic Acid