Three strains of fructophilic lactic acid bacteria were isolated from flowers in South Africa. The isolates formed a subcluster in the Lactobacillus buchneri phylogenetic group, closely related to Lactobacillus fructivorans, Lactobacillus homohiochii, Lactobacillus lindneri and Lactobacillus sanfranciscensis according to phylogenetic analysis based on the 16S rRNA gene sequences. Levels of DNA-DNA relatedness indicated that the three strains belonged to the same taxon and formed a genetically distinct group, well separated from their phylogenetic relatives. The three strains produced acids from only two of the 49 carbohydrates tested, i.e. D-glucose and Dfructose. D-Fructose was more rapidly fermented than D-glucose. Good growth was recorded on D-fructose or D-glucose in the presence of external electron acceptors. However, delayed growth was recorded on D-glucose without electron acceptors. The novel strains produced lactic acid, ethanol and acetic acid from D-glucose at a ratio of 1 : 0.8 : 0.2. These characteristics were distinct from other species of the genus Lactobacillus. Based on the data provided, the three isolates represent a fructophilic and novel species of the genus Lactobacillus, for which the name Lactobacillus florum sp. nov. is proposed. The type strain is F9-1 T (5JCM 16035 T 5DSM 22689 T 5NRIC 0771 T ).Fructophilic lactic acid bacteria (LAB) are a group of LAB described recently (Endo & Okada, 2008). These organisms are known to have unique characteristics, i.e. they grow well on D-fructose or D-glucose in the presence of external electron acceptors, but grow poorly on D-glucose without electron acceptors (Endo & Okada, 2008). Pyruvate and oxygen are used as electron acceptors. At the time of writing, only five species with these characteristics have been described, Fructobacillus fructosus, Fructobacillus durionis, Fructobacillus ficulneus and Fructobacillus pseudoficulneus (Endo & Okada, 2008) and Lactobacillus kunkeei (Endo et al., 2009).During a study of fructophilic LAB, three strains were isolated from flowers in South Africa (Endo et al., 2009). The isolates shared several common characteristics with known fructophilic LAB, but also had a few different characteristics. They formed a subcluster within the Lactobacillus buchneri cluster on phylogenetic analysis based on their 16S rRNA gene sequences and were closely related to Lactobacillus lindneri. Levels of DNA2DNA similarity and biochemical characteristics showed clear separation of the isolates from L. lindneri. This paper describes the taxonomic study of the three novel isolates and their classification as a novel species of the genus Lactobacillus. The three novel strains, F9-1 T , F9-2 and F17, grew well in FYP broth, but poorly in GYP broth and were thus regarded as fructophilic LAB. They were preserved at 280 u C in nutrient broth containing 20 % (v/v) glycerol.The 16S rRNA gene sequences of the three isolates were determined according to a previously described method (Endo & Okada, 2005). The closest recognized relatives of t...