Lactobacillus plajomi sp. nov. and Lactobacillus modestisalitolerans sp. nov., isolated from traditional fermented foods.

Three Lactobacillus-like strains, NB53T, NB446T and NB702, were isolated from traditional fermented food in Thailand. Comparative 16S rRNA gene sequence analysis indicated that these strains belong to the Lactobacillus plantarum group. Phylogenetic analysis based on the dnaK, rpoA, pheS and recA gene sequences indicated that these three strains were distantly related to known species present in the L. plantarum group. DNA-DNA hybridization with closely related strains demonstrated that these strains represented two novel species; the novel strains could be differentiated based on chemotaxonomic and phenotypic characteristics. Therefore, two novel species of the genus Lactobacillus, Lactobacillus plajomi sp. nov. (NB53T) and Lactobacillus modestisalitolerans sp. nov. (NB446T and NB702), are proposed with the type strains NB53T ( = NBRC 107333T = BCC 38054T) and NB446T ( = NBRC 107235T = BCC 38191T), respectively.

Neokomagataea gen. nov., with descriptions of Neokomagataea thailandica sp. nov. and Neokomagataea tanensis sp. nov., osmotolerant acetic acid bacteria of the α-Proteobacteria.

Isolates AH11(T) and AH13(T) were isolated from flowers of lantana and candle bush respectively collected in Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates formed an independent cluster, which was then connected to the type strain of Saccharibacter floricola. The calculated pair-wise 16S rRNA gene sequence similarities of isolate AH11(T) were 95.7-92.3% to the type strains of the type species of the 12 genera of acetic acid bacteria. The DNA base composition was from 51.2 to 56.8 mol % G+C, with a range of 5.6 mol %. When isolate AH11(T) was labeled, DNA-DNA similarities were 100, 12, 4, 5, and 4% respectively to isolates AH11(T) and AH13(T) and the type strains of Saccharibacter floricola, Gluconobacter oxydans, and Acetobacter aceti. The two isolates were non-motile and did not oxidize either acetate or lactate. No growth was found in the presence of 0.35% acetic acid w/v. The two isolates were not osmophilic but osmotolerant, produced 2,5-diketo-D-gluconate from D-glucose, and did not oxidize lactate, thus differing from strains of Saccharibacter floricola, which showed weak lactate oxidation. The two isolates contained unsaturated C(18:1)ω7c fatty acid as the major fatty acid, and were unique in the presence of a considerable amount of straight-chain C(18:1)2OH fatty acid. Q-10 was present as the major isoprenoid quinone. Neokomagataea gen. nov. was proposed with the two species, Neokomagataea thailandica sp. nov. for isolate AH11(T) (=BCC 25710(T)=NBRC 106555(T)), which has 56.8 mol % G+C, and Neokomagataea tanensis sp. nov. for isolate AH13(T) (=BCC 25711(T)=NBRC 106556(T)), which has 51.2 mol % G+C.

Planotetraspora kaengkrachanensis sp. nov. and Planotetraspora phitsanulokensis sp. nov., isolated from soil.

Two novel bacterial strains were isolated from tropical rain forest soil from Thailand. Strains A-T 0875(T) and A-T 1383(T) stained Gram-positive and were filamentous bacteria that developed cylindrical sporangia containing four oval- to rod-shaped spores at the ends of short sporangiophores on branched aerial mycelium. The cell-wall peptidoglycan contained meso-diaminopimelic acid, glutamic acid and alanine as cell-wall amino acids; whole-cell hydrolysates contained rhamnose, madurose, glucose, galactose and 3-O-methylmannose 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-methyl-C(17 : 0). For both strains, the G+C content of the genomic DNA was 71 mol%. Phenotypic and chemotaxonomic analyses showed that the characteristics of the two isolates were typical of members of the genus Planotetraspora. Furthermore, 16S rRNA gene sequence analysis also indicated that the strains belonged to the genus Planotetraspora but as representatives of two novel species. Following an evaluation of our phenotypic, chemotaxonomic and genotypic studies, two novel species are proposed, Planotetraspora kaengkrachanensis sp. nov. (type strain A-T 0875(T)=BCC 24832(T)=NBRC 104272(T)) and Planotetraspora phitsanulokensis sp. nov. (type strain A-T 1383(T)=BCC 26045(T)=NBRC 104273(T)).

Ameyamaea chiangmaiensis gen. nov., sp. nov., an acetic acid bacterium in the alpha-Proteobacteria.

Two isolates, AC04(T) and AC05, were isolated from the flowers of red ginger collected in Chiang Mai, Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates were included within a lineage comprised of the genera Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Neoasaia, Granulibacter, and Tanticharoenia, and they formed an independent cluster along with the type strain of Tanticharoenia sakaeratensis. The calculated pair-wise sequence similarities of isolate AC04(T) were 97.8-92.5% to the type strains of the type species of the 11 genera of acetic acid bacteria. The DNA base composition was 66.0-66.1 mol % G+C with a range of 0.1 mol %. A single-stranded, labeled DNA from isolate AC04(T) presented levels of DNA-DNA hybridization of 100, 85, 4, and 3% respectively to DNAs from isolates AC04(T) and AC05 and the type strains of Tanticharoenia sakaeratensis and Gluconacetobacter liquefaciens. The two isolates were unique morphologically in polar flagellation and physiologically in intense acetate oxidation to carbon dioxide and water and weak lactate oxidation. The intensity in acetate oxidation almost equaled that of the type strain of Acetobacter aceti. The two isolates had Q-10. Isolate AC04(T) was discriminated from the type strains of the type species of the 11 genera by 16S rRNA gene restriction analysis using restriction endonucleases TaqI and Hin6I. The unique phylogenetic, genetic, morphological, physiological, and biochemical characteristics obtained indicate that the two isolates can be classified into a separate genus, and Ameyamaea chiangmaiensis gen. nov., sp. nov. is proposed. The type strain is isolate AC04(T) (=BCC 15744(T), =NBRC 103196(T)), which has a DNA G+C content of 66.0 mol %.

Three new species of bipolar budding yeasts of the genus Hanseniaspora and its anamorph Kloeckera isolated in Thailand.

In the course of a survey of yeast biodiversity in the natural substrates in Thailand, eight strains were found to represent three hitherto undescribed species of Hanseniaspora/Kloeckera. They were isolated from insect frass, flower, lichen, rotted fruit and rotted wood. Based on the morphological and physiological characteristics, and sequences of D1/D2 domain, six strains represent a single species of the genus Hanseniaspora, described as Hanseniaspora thailandica sp. nov. (type BCC 14938(T)=NBRC 104216(T)=CBS 10841(T)), and another strain as Hanseniaspora singularis sp. nov. (type BCC 15001(T)=NBRC 104214(T)=CBS 10840(T)). A further strain, which belongs to Kloeckera and does not produce ascospores, is described as Kloeckera hatyaiensis sp. nov. (type BCC 14939(T)=NBRC 104215(T)=CBS 10842(T)). Strains belonging to H. thailandica sp. nov. differed by 17-19 nucleotide substitutions from Hanseniaspora meyeri, the closest species. DNA reassociation between the two taxa showed 30-48% relatedness. Kloeckera hatyaiensis sp. nov. and H. singularis sp. nov. differed by eight and 16 nucleotide substitutions with one gap from the nearest species, Hanseniaspora clermontiae and Hanseniaspora valbyensis, respectively.

Planotetraspora thailandica sp. nov., isolated from soil in Thailand.

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-O-methylmannose 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 71.1 mol%. The phenotypic and chemotaxonomic analyses showed that the isolate had characteristics typical of members of the genus Planotetraspora. Furthermore, 16S rRNA gene sequence analysis also indicated that this strain belongs to the genus Planotetraspora, but as a putative novel species. Following phenotypic, chemotaxonomic and genotypic studies, the isolate is proposed to be a representative of a novel species, to be named Planotetraspora thailandica sp. nov. The type strain is BCC 21825(T) (=NBRC 104271(T)). An emended description of the genus Planotetraspora is also presented.

Gluconobacter japonicus sp. nov., an acetic acid bacterium in the Alphaproteobacteria.

Five strains, NBRC 3271(T), NBRC 3272, NBRC 3263, NBRC 3260 and NBRC 3269 were examined genetically, phylogenetically, phenotypically and chemotaxonomically. The DNA G+C contents of the five strains were 55.1-56.4 mol%. The five strains had low levels of DNA-DNA hybridization of 13-51 % to the type strains of Gluconobacter frateurii, Gluconobacter thailandicus, Gluconobacter oxydans, Gluconobacter cerinus, Gluconobacter albidus and Gluconobacter kondonii and formed a cluster that was separate from the type strains of the six Gluconobacter species given above in phylogenetic trees based on 16S rRNA gene and 16S-23S rRNA gene internal transcribed spacer sequences. The five strains weakly produced dihydroxyacetone from glycerol, but not 2,5-diketo-d-gluconate or a water-soluble brown pigment from d-glucose and contained ubiquinone-10. The five strains were assigned as representing a novel species of the genus Gluconobacter, for which the name Gluconobacter japonicus sp. nov. is proposed. The type strain is NBRC 3271(T) (=BCC 14458(T)=strain 7(T), K. Kondo). Cells of the type strain are motile by means of polar flagella and the DNA G+C content is 56.4 mol%.

Candida kanchanaburiensis sp. nov., a new ascomycetous yeast species related to Pichia nakazawae isolated in Thailand.

Three yeast strains, ST-633, ST-634 and ST-635, isolated from the fruit body of a mushroom, Coprinus sp., and rotted fruit of guava collected in the western region of Thailand, were found to represent a hitherto undescribed species. This yeast is related to Pichia nakazawae var. akitaensis, P. nakazawae var. nakazawae and Pichia philogaea in the D1/D2 domain of 26S rDNA but 12 (2.3%), 13 (2.5%) and 15 (2.8%) nucleotides are different from these taxa, respectively, suggesting the distinctness of the Thai strains at species level. Since ascospore formation was not detected, it is described as a new species of Candida, Candida kanchanaburiensis. This species is distinguished from P. nakazawae by the ability to assimilate 2-ketogluconic acid and L-lysine, and inability to assimilate soluble starch.

Gluconobacter sphaericus (Ameyama 1975) comb. nov., a brown pigment-producing acetic acid bacterium in the Alphaproteobacteria.

Strain NBRC 12467T was examined genetically, phylogenetically, phenotypically, and chemotaxonomically. The DNA G+C content of the strain was 59.5 mol%. The strain represented low levels of DNA-DNA hybridization of 49-9% to the type strains of eight Gluconobacter species. The strain formed a cluster along with the type strains of G. albidus and G. kondonii in phylogenetic trees based on 16S rRNA gene sequences. In a phylogenetic tree based on 16S-23S rRNA gene ITS sequences, however, the strain formed an independent cluster from the type strains of the eight Gluconobacter species. Such phylogenetic relationships were supported by the calculated pair-wise 16S rRNA gene and 16S-23S rRNA gene ITS sequence similarities. The strain was distinguished from the type strains of the eight Gluconobacter species by 16S-23S rRNA gene ITS restriction analysis using five restriction endonucleases. The strain produced a water-soluble brown pigment and 2,5-diketo-D-gluconate from D-glucose, differing from the type strains of the eight Gluconobacter species, and acid from meso-erythritol very weakly, differing from the type strains of the remaining seven Gluconobacter species except for the type strain of G. roseus, but not from maltose, differing from the type strain of G. oxydans, and had Q-10. For the strain, which was once classified as G. oxydans subsp. sphaericus, Gluconobacter sphaericus (Ameyama 1975) comb. nov. is proposed. The type strain is NBRC 12467T, which is also deposited as BCC 14448T.

Candida ratchasimensis sp. nov. and Candida khaoyaiensis sp. nov., two anamorphic yeast species isolated from flowers in Thailand.

Two yeast strains of the genus Candida were isolated from wild flowers collected in Khao Yai National Park, Nakhonratchasima Province, Thailand. Based on morphological, physiological and chemotaxonomic characteristics and sequence analysis of the D1/D2 domain of the 26S rRNA gene, strains BCC 7722(T) (=NBRC 102563(T)=CBS 10611(T)) and BCC 7729(T) (=NBRC 102565(T)=CBS 10839(T)) were found to represent two distinct novel Candida species, for which the names Candida ratchasimensis sp. nov. and Candida khaoyaiensis sp. nov. are proposed, respectively. In the phylogenetic tree constructed according to the neighbour-joining method based on sequences of the D1/D2 domain of the 26S rRNA gene, strains BCC 7722(T) (GenBank accession no. AY228492) and BCC 7729(T) (accession no. DQ400367) constituted a cluster with Candida cellae that was connected with a clade with Starmerella meliponinorum and Candida etschellsii. Within the D1/D2 domain, C. ratchasimensis and C. khaoyaiensis differ from C. cellae in 25 nucleotide substitutions with five gaps and 29 nucleotide substitutions with one gap, respectively.

Two new ascomycetous anamorphic yeast species related to Candida friedrichii--Candida jaroonii sp. nov., and Candida songkhlaensis sp. nov.--isolated in Thailand.

In a study of yeast diversity in Thailand, eight strains of hitherto undescribed anamorphic yeasts were isolated: four from insect frass, two from Marasmius sp. fruiting bodies, one from a flower, and one from jackfruit exudates. Phylogenetic analysis of the D1/D2 domain of 26S ribosomal DNA nucleotide sequences indicated that the eight strains represented two new species related to Candida friedrichii. Genetic separation of the two new species was further supported by DNA-DNA hybridization analysis, which resulted in between-species similarity values of less than 48%, and by electrophoretic karyotyping. The two new species are C. jaroonii sp. nov. (type strain, ST-300(T) = NBRC 103209(T) = BCC 11783(T) = CBS 10790(T)) and C. songkhlaensis sp. nov. (type strain, ST-328(T) = NBRC 103214(T) = BCC 11804(T) = CBS 10791(T)).

Asaia lannaensis sp. nov., a new acetic acid bacterium in the Alphaproteobacteria.

Asaia lannaensis sp. nov. was described for two strains isolated from flowers of the spider lily collected in Chiang Mai, Thailand. The isolates produced acetic acid from ethanol on ethanol/calcium carbonate agar, differing from the type strains of Asaia bogorensis, Asaia siamensis, and Asaia krungthepensis, but did not grow in the presence of 0.35% acetic acid (v/v). The new species is the fourth of the genus Asaia, the family Acetobacteraceae.

Tanticharoenia sakaeratensis gen. nov., sp. nov., a new osmotolerant acetic acid bacterium in the alpha-Proteobacteria.

Tanticharoenia sakaeratensis gen. nov., sp. nov. is proposed for three strains isolated from soil collected in Thailand. The three strains, AC37(T), AC38, and AC39, were included within a lineage comprising the genera Asaia, Kozakia, Swaminathania, Neoasaia, Acetobacter, Gluconobacter, and Saccharibacter in a phylogenetic tree based on 16S rRNA gene sequences, but formed a quite different, independent cluster. Pair-wise sequence similarities of strain AC37(T) were 96.5-92.1% to the type strains of Acetobacter aceti, Gluconobacter oxydans, Acidomonas methanolica, Gluconacetobacter liquefaciens, Asaia bogorensis, Kozakia baliensis, Swaminathania salitolerans, Saccharibacter floricola, Neoasaia chiangmaiensis, and Granulibacter bethesdensis. The three strains had DNA base compositions comprising respectively 65.6, 64.5, and 65.6 mol % G+C with a range of 1.1 mol %, and formed a single species. Phenotypically, the three strains did not oxidize acetate or lactate, but grew on 30% D-glucose (w/v). Chemotaxonomically, they had Q-10. The type strain is AC37(T) (= BCC 15772(T) = NBRC 103193(T)).

Two new anamorphic yeasts, Candida thailandica sp. nov. and Candida lignicola sp. nov., isolated from insect frass in Thailand.

Two new yeast strains of the genus Candida were isolated from insect frass collected in Khao-Yai National Park, Nakhonrachasima, Thailand. Based on the morphological, physiological and chemotaxonomic characteristics, and sequence analysis of the D1/D2 domain of 26S rRNA gene, these two strains were found to represent two distinct undescribed species and were named Candida thailandica sp. nov. (ST-17 = BCC 7717(T) = NBRC 102562(T)=CBS 10 610) and Candida lignicola sp. nov. (ST-33 = BCC 7733(T) = NBRC 102564(T) = CBS 10612). In the D1/D2 domain of 26S rRNA gene, C. thailandica (GeneBank accession no. AY228491) differs from Candida tsuchiyae, the nearest species, in 66 nucleotide substitutions (10%) and C. lignicola (GeneBank accession no. AY845350) differs from Candida coipomoensis, the nearest species, in nine nucleotides (1.6%). These two new species are clearly distinguished from their closest species by the assimilation of several carbon compounds.

Candida kazuoi sp. nov. and Candida hasegawae sp. nov., two new species of ascomycetous anamorphic yeasts isolated from insect frass in Thailand.

Two strains of anamorphic yeasts isolated from insect frass collected in southern Thailand were assigned to the genus Candida based on the conventional taxonomic criteria used for yeast classification. In the phylogenetic tree based on the D1/D2 domain of the 26S rDNA, these strains are distant from the known species of yeasts and considered to represent two different new species. They are named Candida kazuoi sp. nov. and Candida hasegawae sp. nov.