Lactobacillus bobalius sp. nov., a lactic acid bacterium isolated from Spanish Bobal grape must

1 ENOLAB – Laboratorio de Microbiología Enológica, Departamento de Microbiología y Ecología, Facultad de Biología, Universidad de Valencia, Dr Moliner 50, E-46100 Burjassot, Valencia, Spain 2 Institut Mediterrani d'Estudis Avançats (CSIC-UIB), E-07190 Esporles, Mallorca, Spain Correspondence Isabel Pardo Isabel.Pardo{at}uv.es A Lactobacillus strain, designated 203 T , previously isolated from Bobal grape must was characterized phylogenetically, genotypically and phenotypically in order to establish whether it represents a novel species. On the basis of the 16S rRNA gene sequence, strain 203 T was shown to belong to the genus Lactobacillus , falling within the Lactobacillus alimentarius–Lactobacillus farciminis group and being closely related to the type strains of L. alimentarius , Lactobacillus kimchii and Lactobacillus paralimentarius . DNA–DNA hybridization results confirmed the separate status of strain 203 T at the species level. To establish the similarities and differences between 203 T and the three aforementioned closest species, the following methods were used: amplified rDNA restriction analysis, analysis of the 16S–23S rDNA intergenic spacer region, random amplification of polymorphic DNA (RAPD) profiling, ribotyping, carbohydrate fermentation and physiological tests. Strain 203 T could be differentiated genetically using RAPD analysis and ribotyping. Phenotypically, it can be distinguished from its closest relatives by its ability to grow at pH 3.3, by gas production from gluconate and by certain carbohydrate fermentations. On the basis of these data, strain 203 T represents a novel species of the genus Lactobacillus , for which the name Lactobacillus bobalius sp. nov. is proposed. The type strain is 203 T (=CECT 7310 T =DSM 19674 T ). Abbreviations: ARDRA, amplified rDNA restriction analysis; ISR, intergenic spacer region; RAPD, random amplification of polymorphic DNA The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain 203 T is AY681134 . Neighbour-joining and maximum-parsimony phylogenetic trees for strain 203 T and members of the genus Lactobacillus are available as supplementary material with the online version of this paper.