Five bacterial strains were isolated from tropical flowers collected in Thailand and Indonesia by the enrichment culture approach for acetic acid bacteria. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates were located within the cluster of the genus Asaia. The isolates constituted a group separate from Asaia bogorensis on the basis of DNA relatedness values. Their DNA GMC contents were 586-597 mol %, with a range of 11 mol %, which were slightly lower than that of A. bogorensis (593-610 mol %), the type species of the genus Asaia. The isolates had morphological, physiological and biochemical characteristics similar to A. bogorensis strains, but the isolates did not produce acid from dulcitol. On the basis of the results obtained, the name Asaia siamensis sp. nov. is proposed for these isolates. Keywords : Asaia siamensis sp. nov., acetic acid bacteria, Acetobacteraceae, ProteobacteriaThe genus Asaia was introduced with a single species, Asaia bogorensis, in the family Acetobacteraceae (Yamada et al., 2000). In contrast with strains of the genera Acetobacter, Gluconobacter and Gluconacetobacter, the strains assigned to this genus are characterized by no or very weak capability for oxidizing ethanol to acetic acid and no growth in the presence of acetic acid (0n35 %, v\v). This paper describes the proposal of Asaia siamensis sp. nov., the second species of the genus Asaia, for strains isolated from tropical flowers collected in Thailand and Indonesia. Five bacterial strains (S60-1 T , D4-1, Y85, i36 and B28S-3) were isolated from tropical flowers collected in Thailand and Indonesia by the enrichment culture approach using a sorbitol medium and a dulcitol medium at pH 3n5 ( These strains were maintained on agar slants of AG medium composed of 0n1% -glucose, 1n5 % glycerol, 0n5 % peptone, 0n5 % yeast extract, 0n2 % malt extract, 0n7 % CaCO $ and 1n5 % agar (w\v). Acetobacter aceti IFO 14818 T , Gluconobacter oxydans IFO 14819 T , Gluconacetobacter liquefaciens IFO 12388 T and Asaia bogorensis JCM 10569 T , NRIC 0317 and NRIC 0318 were used as reference strains. Morphological, physiological and biochemical characteristics were examined according to the methods reported by Asai et al. (1964) and Yamada et al. (1976Yamada et al. ( , 1999Yamada et al. ( , 2000. Cells of all the isolates were Gramnegative, strictly aerobic and rod-shaped, measuring 0n6-1n0i1n0-4n5 µm. The cells were motile by means of peritrichous flagella. Colonies were pink, shiny, smooth and raised with an entire margin on AG agar plates. All the isolates grew well at pH 3n0 and 3n5 and at 30 mC on a CaCO $ -free AG medium.

Three bacterial strains were isolated from flowers collected in Bangkok, Thailand, by an enrichment-culture approach for acetic acid bacteria. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates were located in the lineage of the genus Asaia but constituted a cluster separate from the type strains of Asaia bogorensis and Asaia siamensis. The DNA base composition of the isolates was 60?2-60?5 mol% G+C, with a range of 0?3 mol%. The isolates constituted a taxon separate from Asaia bogorensis and Asaia siamensis on the basis of DNA-DNA relatedness. The isolates had morphological, physiological, biochemical and chemotaxonomic characteristics similar to those of the type strains of Asaia bogorensis and Asaia siamensis, but the isolates grew on maltose. The major ubiquinone was Q 10 . On the basis of the results obtained, the name Asaia krungthepensis sp. nov. is proposed for the isolates. The type strain is isolate AA08 T (=BCC 12978 T =TISTR 1524 T =NBRC 100057 T =NRIC 0535 T ), which had a DNA G+C content of 60?3 mol% and was isolated from a heliconia flower ('paksaasawan' in Thai; Heliconia sp.) collected in Bangkok, Thailand.The genus Asaia was introduced with a single species, Asaia bogorensis, as the fifth genus of the family Acetobacteraceae (Yamada et al., 2000). Asaia siamensis, the second species, was described by Katsura et al. (2001). Strains assigned to the genus Asaia are characterized by poor or non-existent production of acetic acid from ethanol and by the absence of growth in the presence of 0?35 % acetic acid (v/v). On the other hand, strains of the genera Acetobacter, Gluconobacter, Acidomonas, Gluconacetobacter and Kozakia produce large amounts of acetic acid from ethanol and grow in the presence of 0?35 % acetic acid. According to Yamada et al. (2000) and Katsura et al. (2001), strains of the genus Asaia have been isolated mostly from tropical flowers. During the course of our studies on acetic acid bacteria isolated from flowers collected in Thailand, we found that three isolates constitute a novel species; this paper describes Asaia krungthepensis sp. nov.Three bacterial strains were isolated from heliconia flowers ('paksaasawan' in Thai; Heliconia sp.) collected in Bangkok, Thailand, by an enrichment-culture approach using a sucrose medium, which was composed of 2?0 % sucrose, 0?3 % peptone, 0?3 % yeast extract and 0?3 % acetic acid (v/v) and adjusted to pH 3?5. When bacterial growth was found in the sucrose medium, micro-organisms were streaked on agar plates containing 2?0 % D-glucose, 0?5 % ethanol (v/v), 0?3 % peptone, 0?3 % yeast extract, 0?7 % CaCO 3 and 1?2 % agar (w/v). These acetic acid bacteria were selected as acid-producing bacterial strains that formed clear zones around colonies on agar plates. The three isolates, AA08 T (=BCC 12978 T ), AA09 (=BCC 12979) and AA06 (=BCC 12977), were cultured at 30 uC on agar slants comprising glucose/ethanol/CaCO 3 /agar medium (GECA medium) that contained 2?5 % D-glucose, 0?5 % ethanol, 3Present address: 2-3-21 Seinan...

A Gram-positive-staining, filamentous bacterial strain that developed cylindrical sporangia containing four oval-to rod-shaped spores at the ends of short sporangiophores on branched aerial mycelium was isolated from tropical rainforest soil near a hot spring. The cell-wall peptidoglycan contained meso-diaminopimelic acid, glutamic acid and alanine as cell-wall amino acids; the whole-cell hydrolysate contained rhamnose, madurose, glucose, galactose and 3-Omethylmannose as whole-cell sugars. The predominant menaquinone was MK-9(H 4 ). Mycolic acids were not detected. The diagnostic phospholipid was phosphatidylethanolamine. The predominant cellular fatty acids were iso-C 16 : 0 and 10-methylated C 17 : 0 . The G+C content of the DNA was 71

Neoasaia chiangmaiensis gen. nov., sp. nov., a novel osmotolerant acetic acid bacterium in the a a-Proteobacteria