Genome-based Reclassification of Paraburkholderia insulsa as a Later Heterotypic Synonym of Paraburkholderia fungorum and Proposal of Paraburkholderia terrae subsp. terrae subsp. nov. and Paraburkholderia terrae subsp. steynii subsp. nov.

Based on the 16S rRNA gene sequences similarity of > 99.8%, the phylogeny of 88 Paraburkholderia strains was reconstructed. Further, they were subjected to overall genome-related indices (OGRI), which resulted in the identification of distinct pairs of species that were closely related. A pair consist of the type strains of Paraburkholderia insulsa and Paraburkholderia fungorum possessed a dDDH value of 87.9%, correspondingly, and the average nucleotide identity (ANI) value was 98.5%. Based on the phylogenetic analysis, OGRI and phenotypical evidence, P. insulsa was proposed as a later heterotypic synonym of P. fungorum. Furthermore, a pair comprising type strains of Paraburkholderia terrae and Paraburkholderia steynii possessed dDDH and ANI values of 71.2% and 96.6%, respectively, and difference in phenotypic traits, which supports a subspecies proposal within these taxa. Thus, the recently described Paraburkholderia steynii was proposed into two subspecies namely Paraburkholderia terrae subsp terrae subsp. nov and as Paraburkholderia terrae subsp. steynii subsp. nov.

In vitro and in silico analysis of Brilliant Black degradation by Actinobacteria and a Paraburkholderia sp.

The soil bacteria isolated in this study, including three strains of actinobacteria and one Paraburkholderia sp., showed decolorization activity of azo dyes in the resting cell assay and were shown to use methyl red as the sole carbon source to proliferate. Therefore, their ability to degrade, bioabsorb, or a combination of both mechanism was investigated using the substrate brilliant black. The strains DP-A9 and DP-L11, within 24 h of incubation, showed complete biodegradation of 173.54 mg/L brilliant black and the strains DP-D10 and DP-P12 showed partial decolorization of 83.3 mg/L and 36.4 mg/L, respectively, by both biosorption and biodegradation. In addition, the shotgun assembled genome of these strains showed a highly diverse set of genes encoding for candidate dye degrading enzymes, providing avenues to study azo dye metabolism in more detail.

Proposal of Carbonactinosporaceae fam. nov. within the class Actinomycetia. Reclassification of Streptomyces thermoautotrophicus as Carbonactinospora thermoautotrophica gen. nov., comb. nov.

Streptomyces thermoautotrophicus UBT1T has been suggested to merit generic status due to its phylogenetic placement and distinctive phenotypes among Actinomycetia. To evaluate whether 'S. thermoautotrophicus' represents a higher taxonomic rank, 'S. thermoautotrophicus' strains UBT1T and H1 were compared to Actinomycetia using 16S rRNA gene sequences and comparative genome analyses. The UBT1T and H1 genomes each contain at least two different 16S rRNA sequences, which are closely related to those of Acidothermus cellulolyticus (order Acidothermales). In multigene-based phylogenomic trees, UBT1T and H1 typically formed a sister group to the Streptosporangiales-Acidothermales clade. The Average Amino Acid Identity, Percentage of Conserved Proteins, and whole-genome Average Nucleotide Identity (Alignment Fraction) values were ≤58.5%, ≤48%, ≤75.5% (0.3) between 'S. thermoautotrophicus' and Streptosporangiales members, all below the respective thresholds for delineating genera. The values for genomics comparisons between strains UBT1T and H1 with Acidothermales, as well as members of the genus Streptomyces, were even lower. A review of the 'S. thermoautotrophicus' proteomic profiles and KEGG orthology demonstrated that UBT1T and H1 present pronounced differences, both tested and predicted, in phenotypic and chemotaxonomic characteristics compared to its sister clades and Streptomyces. The distinct phylogenetic position and the combination of genotypic and phenotypic characteristics justify the proposal of Carbonactinospora gen. nov., with the type species Carbonactinospora thermoautotrophica comb. nov. (type strain UBT1T, = DSM 100163T = KCTC 49540T) belonging to Carbonactinosporaceae fam. nov. within Actinomycetia.

Phylogenomic analyses of the Staphylococcaceae family suggest the reclassification of five species within the genus Staphylococcus as heterotypic synonyms, the promotion of five subspecies to novel species, the taxonomic reassignment of five Staphylococcus species to Mammaliicoccus gen. nov., and the formal assignment of Nosocomiicoccus to the family Staphylococcaceae.

Phylogenetic analyses based on 16S rRNA gene sequences of members of the family Staphylococcaceae showed the presence of para- and polyphyletic genera. This finding prompted a thorough investigation into the taxonomy of the Staphylococcaceae family by analysing their core genome phylogeny complemented with genome-based indices such as digital DNA-DNA hybridization, average nucleotide identity and average amino acid identity. The resulting data suggested the following proposals: Auricoccus indicus was reduced in taxonomic rank as a later heterotypic synonym of Abyssicoccus albus; Staphylococcus petrasii subsp. jettensis as a later heterotypic synonym of Staphylococcus petrasii subsp. petrasii; the unification of Staphylococcus aureus subsp. anaerobius and Staphylococcus aureus subsp. aureus as Staphylococcus aureus; the unification of Staphylococcus carnosus subsp. utilis and Staphylococcus carnosus subsp. carnosus as Staphylococcus carnosus; the unification of Staphylococcus saprophyticus subsp. bovis and Staphylococcus saprophyticus subsp. saprophyticus as Staphylococcus saprophyticus; Staphylococcus succinis subsp. casei as the novel species Staphylococcus casei; Staphylococcus schleiferi subsp. coagulans as the novel species Staphylococcus coagulans; Staphylococcus petrasii subsp. croceilyticus as the novel species Staphylococcus croceilyticus; Staphylococcus petrasii subsp. pragensis as the novel species Staphylococcus pragensis; Staphylococcus cohnii subsp. urealyticus as the novel species Staphylococcus urealyticus; the reassignment of Staphylococcus sciuri, Staphylococcus fleurettii, Staphylococcus lentus, Staphylococcus stepanovicii and Staphylococcus vitulinus to the novel genus Mammaliicoccus with Mammaliicoccus sciuri as the type species; and the formal assignment of Nosocomiicoccus as a member of the family Staphylococcaceae.

Sphingomonas palmae sp. nov. and Sphingomonas gellani sp. nov., endophytically associated phyllosphere bacteria isolated from economically important crop plants.

In this study, two endophytic bacterial strains designated JS21-1T and S6-262T isolated from leaves of Elaeis guineensis and stem tissues of Jatropha curcas respectively, were subjected for polyphasic taxonomic approach. On R2A medium, colonies of strains JS21-1T and S6-262T are orange and yellow, respectively. Phylogenetic analyses using 16S rRNA gene sequencing and whole-genome sequences placed the strains in distinct clades but within the genus Sphingomonas. The DNA G + C content of JS21-1T and S6-262T were 67.31 and 66.95%, respectively. Furthermore, the average nucleotide identity and digital DNA-DNA hybridization values of strains JS21-1T and S6-262T with phylogenetically related Sphingomonas species were lower than 95% and 70% respectively. The chemotaxonomic studies indicated that the major cellular fatty acids of the strain JS21-1T were summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), C16:0, and C14:0 2OH; strain S6-262T possessed summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH) and summed feature 8 (C18:1 ω6c and/or C18:1 ω7c). The major quinone was Q10, and the unique polyamine observed was homospermidine. The polar lipid profile comprised of mixture of sphingoglycolipid, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and certain uncharacterised phospholipids and lipids. Based on this polyphasic evidence, strains JS21-1T and S6-262T represent two novel species of the genus Sphingomonas, for which the names Sphingomonas palmae sp. nov. and Sphingomonas gellani sp. nov. are proposed, respectively. The type strain of Sphingomonas palmae sp. nov. is JS21-1T (= DSM 27348T = KACC 17591T) and the type strain of Sphingomonas gellani sp. nov. is S6-262T (= DSM 27346T =  KACC 17594T).

Genome-based analyses reveal the presence of 12 heterotypic synonyms in the genus Streptomyces and emended descriptions of Streptomyces bottropensis, Streptomyces celluloflavus, Streptomyces fulvissimus, Streptomyces glaucescens, Streptomyces murinus, and Streptomyces variegatus.

Phylogenetic analysis based on 16S rRNA gene sequences of the genus Streptomyces showed the presence of six distinguishable clusters, with 100 % sequence similarity values among strains in each cluster; thus they shared almost the same evolutionary distance. This result corroborated well with the outcome of core gene (orthologous gene clusters) based genome phylogeny analysis of 190 genomes including the Streptomyces species in those six clusters. These preeminent results led to an investigation of genome-based indices such as digital DNA-DNA hybridization (dDDH), average nucleotide identity (ANI) and average amino acid identity (AAI) for the strains in those six clusters. Certain strains recorded genomic indices well above the threshold values (70 %, 95-96 % and >95 % for dDDH, ANI and AAI, respectively) determined for species affiliation, suggesting only one type strain belongs to described species and the other(s) may need to be reduced in taxa to a later heterotypic synonym. To conclude, the results of comprehensive analyses based on phylogenetic and genomic indices suggest that the following six reclassifications are proposed: Streptomyces flavovariabilis as a later heterotypic synonym of Streptomyces variegatus; Streptomyces griseofuscus as a later heterotypic synonym of Streptomyces murinus; Streptomyces kasugaensis as a later heterotypic synonym of Streptomyces celluloflavus; Streptomyces luridiscabiei as a later heterotypic synonym of Streptomyces fulvissimus; Streptomyces pharetrae as a later heterotypic synonym of Streptomyces glaucescens; and Streptomyces stelliscabiei as a later heterotypic synonym of Streptomyces bottropensis.

Chitinasiproducens palmae gen. nov., sp. nov., a new member of the family Burkholderiaceae isolated from leaf tissues of oil palm (Elaeis guineensis Jacq.).

A Gram-stain-negative, aerobic, rod-shaped, leaf-associated bacterium, designated JS23T, was isolated from surface-sterilized leaf tissue of an oil palm grown in Singapore and was investigated by polyphasic taxonomy. Phylogenetic analyses based on 16S rRNA gene sequences and 180 conserved genes in the genome of several members of Burkholderiaceae revealed that strain JS23T formed a distinct evolutionary lineage independent of other taxa within the family Burkholderiaceae. The predominant ubiquinone was Q-8. The primary polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and an unidentified aminophospholipid. The major fatty acids were C16 : 0, summed feature 3 (C16 : 1 ω7c /C16 : 1 ω6c) and summed feature 8 (C18 : 1 ω7c /C18 : 1 ω6c). The size of the genome is 5.36 Mbp with a DNA G+C content of 66.2 mol%. Genomic relatedness measurements such as average nucleotide identity, genome-to-genome distance and digital DNA-DNA hybridization clearly distinguished strain JS23T from the closely related genera Burkholderia, Caballeronia, Mycetohabitans, Mycoavidus, Pandoraea, Paraburkholderia, Robbsia and Trinickia. Furthermore, average amino acid identity values and the percentages of conserved proteins, 56.0-68.4 and 28.2-45.5, respectively, were well below threshold values for genus delineation and supported the assignment of JS23T to a novel genus. On the basis of the phylogenetic, biochemical, chemotaxonomic and phylogenomic evidence, strain JS23T is proposed to represent a novel species of a new genus within the family Burkholderiaceae, for which the name Chitinasiproducens palmae gen. nov., sp. nov., is proposed with the type strain of JS23T (= DSM 27307T=KACC 17592T).

Reclassification of Sphingomonas aeria as a later heterotypic synonym of Sphingomonas carotinifaciens based on whole-genome sequence analysis.

The 16S rRNA gene sequences of Sphingomonas carotinifaciens L9-754T and Sphingomonas aeria B093034T possess 99.71 % sequence similarity. Further studies were undertaken to clarify the taxonomic assignments of these species. Whole-genome comparisons showed that S. aeria B093034Tand S. carotinifaciens L9-754T shared 96.9 % average nucleotide identity, 98.4 % average amino acid identity and 76.1 % digital DNA-DNA hybridization values. These values exceeded or approached the recommended species delineation threshold values. Furthermore, a phylogenetic tree based on 41 of the most conserved genes provided additional evidence that S. aeria B093034T and S. carotinifaciens L9-754T are very closely related. Based on this evidence we propose the reclassification of S. aeria Xue et al. 2018 as a later heterotypic synonym of S. carotinifaciens Madhaiyan et al. 2017.

Phytobacter palmae sp. nov., a novel endophytic, N2 fixing, plant growth promoting Gammaproteobacterium isolated from oil palm (Elaeis guineensis Jacq.).

A novel strain S29T with high nitrogen fixing ability was isolated from surface-sterilized leaf tissues of oil palm (Elaeis guineensis) growing in Science Park II, Singapore. On the basis of 16S rRNA gene analysis and multilocus sequence typing with the rpoB, gyrB, infB and atpD genes, strain S29T was a member of the genus Phytobacter, with Phytobacter ursingii ATCC 27989T and Phytobacter diazotrophicus LS 8T as its closest relatives. Unique biochemical features that differentiated strain S29T from its closest relatives were the ability to utilize melibiose, α-cyclodextrin, glycogen, adonital, d-arabitol, m-inositol and xylitol. The major fatty acids were C14 : 0, C16 : 0, C17 : 0, C16 : 1 ω5c and summed feature 2 containing C16 : 1 ω7c and/or C16 : 1 ω6c. The polar fatty acid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminophospholipid and aminolipids. The draft genome of strain S29T comprised 5, 284, 330 bp with a G + C content of 52.6 %. The average nucleotide identity and digital DNA-DNA hybridization values between strain S29T and the phylogenetically related Enterobacterales species were lower than 95 % and 70 %, respectively. Thus, the polyphasic evidences generated through the phenotypic, chemotaxonomic and genomic methods confirmed that strain S29T represents a novel species of the genus Phytobacter, for which we propose the name Phytobacter palmae sp. nov. with the type strain of S29T (=DSM 27342T=KACC 17598T).

Paenibacillus polysaccharolyticus sp. nov., a xylanolytic and cellulolytic bacteria isolated from leaves of Bamboo Phyllostachys aureosulcata.

A novel xylanolytic and cellulolytic strain, BL9T, was isolated from leaves of the Bamboo plants maintained at the Tamil Nadu Agricultural University Campus, Coimbatore, India. On the basis of the results of 16S rRNA gene analysis, it was determined to be phylogenetically close to the type strains of Paenibacillus amylolyticus NRRL NRS-290T (98.3 %), Paenibacillus barcinonensis BP-23T (98.1 %), Paenibacillus tundrae A10bT (98.0 %) and Paenibacillus xylanexedens B22aT (97.6 %). The strain stained Gram variable and was aerobic, motile and catalase- and oxidase-positive, with rod-shaped cells. Based upon the genome sequence, the average nucleotide identity with the related species ranged from 66 % to 72 %, and the digital DNA-DNA hybridization value ranged from 13 % to 27 %. The DNA G+C content was 45.6 mol%, meso-diaminopimelic acid was identified as the predominant component of the cell wall, and MK-7 was the only menaquinone in cell membranes. The whole-cell fatty acid profile included C16 : 0, iso-C14 : 0, anteiso-C15 : 0 and iso-C16 : 0. The polar lipids identified were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine and diphosphtidylglycerol. On the basis of these polyphasic taxonomic traits, BL9T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus polysaccharolyticus sp. nov. is proposed. The type strain is BL9T (=NBRC 105191T=ICMP 17623T).

Pseudomonas sesami sp. nov., a plant growth-promoting Gammaproteobacteria isolated from the rhizosphere of Sesamum indicum L.

A novel Gram-stain negative, aerobic, motile, rod-shaped bacterium was isolated from Sesame (Sesamum indicum L.) rhizosphere soil. Based on the 16S rRNA gene similarity value (99.4-98.6%) obtained with phylogenetically closely related strains and through analyses of their house keeping genes (atpD, infB and rpoB), the strain SI-P133T was delineated among the species of the genus Pseudomonas and was subjected to polyphasic taxonomic analysis. It was a chemoorganotroph which grew at wide range of temperature (4-45 °C), pH (5.5-9.5) and NaCl concentrations (0-7% (w/v). DNA-DNA hybridization values with closely related type strains DSM 9751T, DSM 19095T, DSM 21509T, ICMP 9151T and DSM 6929T ranged from 23.1 to 44.2%. The most abundant fatty acids were C16:0, C10:0 3-OH, summed feature 3 (comprising C16:1 ω7c and/or C16:1 ω6c), C17:0 cyclo and C12:0 3-OH. The major isoprenoid quinone system was ubiquinone 9 (Q-9) and the G+C content was 61.3 mol%. The major polar lipids of the strain SI-P133T were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. With regard to prospective use in agriculture, plant growth-promoting properties of the strain were tested and plant growth-promotion was demonstrated under in vitro conditions. Based on the various polyphasic taxonomic traits analysed, the strain SI-P-133T was novel and placed within the genus Pseudomonas. Hence we propose a novel species named Pseudomonas sesami sp. nov., for which the type strain is SI-P133T (=NCIMB 14519T = KCTC 22518T).

Paenibacillus methanolicus sp. nov., a xylanolytic, methanol-utilizing bacterium isolated from the phyllosphere of bamboo (Pseudosasa japonica).

Strain BL24T, isolated from bamboo phyllosphere collected in Coimbatore, India, was studied for taxonomic classification. Cells of the strain were aerobic, Gram-stain-positive, motile, catalase- and oxidase-positive rods and grew on media containing methanol. In 16S rRNA gene sequence analysis, strain BL24Tshowed the highest sequence similarities with Paenibacillus phyllosphaeraeKACC 11473T (97.8 %) and Paenibacillus sacheonensisSY01 (95.1 %). DNA-DNA hybridization with P. phyllosphaerae KACC 11473T, phylogenetically the most closely related species, was 21.6 %; this value showed that strain BL24Tbelonged to a different species. The cell-wall peptidoglycan was found to possess meso-diaminopimelic acid and the G+C content of genomic DNA was 52.1 mol %. It contained menaquinone (MK)-7 as the predominant respiratory quinone and the major cellular fatty acids are C16 : 0, anteiso-C15 : 0, iso-C16 : 0, and anteiso-C17 : 0. Based on the molecular and chemotaxonomic markers and physiological properties, strain BL24T (=NRRL B-51698T=CCM 7577T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillusmethanolicusis proposed.

Streptomyces pini sp. nov., an actinomycete isolated from phylloplane of pine (Pinus sylvestris L.) needle-like leaves.

A novel siderophore-producing actinomycete, designated PL19T, was isolated from the Scots-pine needle-like leaves collected from TNAU campus, Coimbatore, India. The isolate was chemoorganotrophic in nutrition and able to grow at 30 °C, and the optimum pH and NaCl facilitated the growth pH 6-11 and 0-8 % (w/v), respectively. The cells are filamentous and the mycelia formed are basically of wide and intricately branched substrate mycelium from which aerial mycelia arises, later gets differentiated into spores that are warty and arranged spirally. The 16S rRNA gene of strain PL19T was sequenced and was highly similar to the type strains of species of the genus Streptomyces, including Streptomyces barkulensis RC1831T (98.8 % pairwise similarity), Streptomyces fenghuangensis GIMN4.003T (98.2 %), Streptomyces nanhaiensis SCSIO 01248T (98.0 %), Streptomyces radiopugnans R97T (97.9 %), Streptomyces atacamensis C60T (97.8 %) and Streptomyces macrosporus NBRC 14749T (97.2 %), all of which were subjected to taxonomical characterization using a polyphasic approach. The strains showed unique carbon utilization patterns, and it possesses iso-C16 : 0 anteiso-C15 : 0 and anteiso-C17 : 0 as a major cellular fatty acids. The cell-wall was dominated with ll-type diaminopimelic acid, and the menaquinone type was MK-9(H6, H8). These chemotaxonomic evidences placed strain PL19T within the genus Streptomyces. The determination of G+C ratio (69.5 mol%) and DNA-DNA hybridization values (13.4-31.8 % with the phylogenetically related species) helped in further hierarchical classification of strain PL19T. Based on morphological, physiological and chemotaxonomic data as well as DNA-DNA hybridization values, strain PL19T could be distinguished from the evolutionarily closest species currently available. All these collective data show that strain PL19T represents a novel species of the genus Streptomyces, for which the name Streptomyces pini sp. nov. is proposed. The type strain is PL19T (=NRRL B-24728T=ICMP 17783T).

Rhodanobacter glycinis sp. nov., a yellow-pigmented gammaproteobacterium isolated from the rhizoplane of field-grown soybean.

A novel, yellow-pigmented bacterium, designated strain MO64(T), was isolated from the rhizoplane of field-grown soybean, collected from an experimental plot at Coimbatore, India. Cells were Gram-reaction-negative, motile, non-spore-forming rods that produced yellow-pigmented colonies on R2A agar. Phylogenetic analysis, based on 16S rRNA gene sequences, showed that strain MO64(T) belonged to the genus Rhodanobacter. Strain MO64(T) was related most closely to Rhodanobacter ginsengisoli GR17-7(T) (98.0% 16S rRNA gene sequence similarity), Rhodanobacter spathiphylli B39(T) (97.9%), Rhodanobacter panaciterrae LnR5-47(T) (97.7%), Rhodanobacter terrae GP18-1(T) (97.6%), Rhodanobacter soli DCY45(T) (97.3%) and Rhodanobacter caeni MJ01(T) (97.2%); levels of similarity to the type strains of all other recognized species of the genus Rhodanobacter were less than 97.0%. Chemotaxonomic data (Q-8 as the predominant ubiquinone, and iso-C(16 : 0), iso-C(15 : 0), C(17 : 0) cyclo, iso-C(17 : 1)ω9c, iso-C(17 : 0) and iso-C(11 : 0) as the major fatty acids) also supported the affiliation of strain MO64(T) with the genus Rhodanobacter. The G+C content of the genomic DNA was 64 mol%. The results of DNA-DNA hybridization and phenotypic analysis showed that strain MO64(T) can be distinguished from all known species of the genus Rhodanobacter and therefore represents a novel species of the genus, for which the name Rhodanobacter glycinis sp. nov. is proposed. The type strain is MO64(T) ( = ICMP 17626(T) = NBRC 105007(T)).

Aggregation of selected plant growth promoting Methylobacterium strains: role of cell surface components and hydrophobicity.

The bacterial cell surface plays a major role in the bacterial aggregation that in turn plays a positive role in affecting the bacterial dispersion and survival in soil and their ability to adhere to plant surfaces. Plant growth-promoting Methylobacterium strains, Methylobacterium goesingense CBMB5, Methylobacterium sp. CBMB12, Methylobacterium oryzae CBMB20, Methylobacterium fujisawaense CBMB37, M. oryzae CBMB110 and Methylobacterium suomiense CBMB120 were evaluated for aggregation efficiency. Aggregation occurred in all test strains under high C/N growth conditions, and the strain CBMB12 showed the highest aggregation of 53.4 % at 72 h. Disaggregation compound treatment studies revealed the role of protein-protein interaction in Methylobacterium strains except CBMB110 and CBMB120 strains, where a possible carbohydrate-protein interaction is suspected. Surface layer protein extraction by LiCl followed by SDS-PAGE analysis showed the presence of proteins at molecular weights ranging from 41 to 49 kDa. Methylobacterium strains under aggregated conditions showed increased hydrophobicity compared to the cells under standard grown conditions. A relatively higher hydrophobicity of 50.1 % as evident by the adhesion with xylene was observed with strain CBMB12 under aggregated condition. This study reports the aggregation ability in plant growth-promoting Methylobacterium strains and the possible involvement of cellular components and hydrophobicity in this phenomenon.

Microbacterium azadirachtae sp. nov., a plant-growth-promoting actinobacterium isolated from the rhizoplane of neem seedlings.

Microbacterium strain AI-S262(T) was isolated from the rhizoplane of neem seedlings in the Botanical garden of Tamilnadu Agricultural University, Coimbatore, India, and subjected to phenotypic, chemotaxonomic and genetic characterization. Cells of this strain were Gram-stain-positive, motile, non-spore-forming, short rods and formed light-yellow-pigmented colonies on nutrient agar. Strain AI-S262(T) contained MK-12 and MK-13 as the main respiratory quinones, anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0) as the predominant fatty acids, peptidoglycan-type B2beta with glycolyl residues, and had a DNA G+C content of 69.5 mol%. A phylogenetic analysis based on 16S rRNA gene sequences showed 98.0-98.6 % pair-wise similarity with respect to close relatives in the genus Microbacterium. DNA-DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 39%) between strain AI-S262(T) and its closest relatives. Data from DNA-DNA hybridization and phenotypic analyses supported the conclusion that strain AI-S262(T) represents a novel species in the genus Microbacterium, for which the name Microbacterium azadirachtae sp. nov. is proposed. The type strain is AI-S262(T) (=JCM 15681(T) =LMG 24772(T) =KCTC 19668(T)).

Enterobacter arachidis sp. nov., a plant-growth-promoting diazotrophic bacterium isolated from rhizosphere soil of groundnut.

A methylotrophic nitrogen-fixing bacterial strain, Ah-143(T), isolated from the rhizosphere soil of field-grown groundnut was analysed by a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence analysis combined with rpoB gene sequence analysis allocated strain Ah-143(T) to the family Enterobacteriaceae, with Enterobacter radicincitans and Enterobacter cowanii as the closest relatives. The strain is Gram-stain-negative, non-spore-forming, aerobic and motile, having straight rod-shaped cells with a DNA G+C content of approximately 53.2 mol%. The strain utilizes methanol as a carbon source and the mxaF gene was closely related to the mxaF gene of members of the genus Methylobacterium. The fatty acid profile consisted of C(16 : 0), C(17 : 0) cyclo, C(18 : 1)omega7c, summed feature 2 (iso-C(16 : 1) I and/or C(14 : 0) 3-OH) and summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) as the major components. DNA-DNA relatedness of strain Ah-143(T) with its close relatives was less than 20 %. On the basis of the phylogenetic analyses, DNA-DNA hybridization data, and unique physiological and biochemical characteristics, it is proposed that the strain represents a novel species of the genus Enterobacter and should be named Enterobacter arachidis sp. nov. The type strain is Ah-143(T) (=NCIMB 14469(T) =KCTC 22375(T)).

Zinc metal solubilization by Gluconacetobacter diazotrophicus and induction of pleomorphic cells.

Gluconacetobacter diazotrophicus strain PAl5 exhibited a minimum inhibitory concentration value of 11 mM in an LGI medium amended with ZnCl2. When an LGI medium was amended with Zn metal, solubilization halos were observed in a plate assay, and further solubilization was confirmed in a broth assay. The maximum solubilization was recorded after 120 h with a 0.1% Zn metal amendment. During solubilization, the culture growth and pH of the broth were indirectly correlated. Using a Fourier Transform Infrared Spectroscopy analysis, one of the agents solubilizing the Zn metal was identified as gluconic acid. When the Zn-amended broth was observed under a bright field microscope, long involution cells were observed, and further analysis with Atomic Force Microscopy revealed highly deformed, pleomorphic, aggregate-like cells.