Halobacillus salinarum sp. nov., Halobacillus shinanisalinarum sp. nov. and Halobacillus amylolyticus sp. nov., isolated from saltern soil.

Three bacterial strains, designated SSBR10-3T, SSTM10-2T and SSHM10-5T, were isolated from saltern soil sampled in Jeollanam-do, Republic of Korea. Cells were aerobic, Gram-stain-positive, flagellated and rod-shaped. The strains grew optimally at 28°C and at pH 7.0. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains SSBR10-3T, SSTM10-2T and SSHM10-5T were placed within the genus Halobacillus, showing the highest similarity to Halobacillus alkaliphilus FP5T (98.6 %), 'Halobacillus ihumii' Marseille-Q1234T (98.5 %) and Halobacillus locisalis MSS-155T (98.6 %), respectively. The genomic similarity values between strains SSBR10-3T, SSTM10-2T and SSHM10-5T and their related species were 17.6-22.6 % for digital DNA-DNA hybridization (dDDH) and 69.6-78.5 % for orthologous average nucleotide identity (OrthoANI), which were lower than the thresholds recommended for species delineation. The dDDH and OrthoANI values among the three strains were below 38.3 and 89.4 %, respectively. Besides the differences in genomic features, strains SSBR10-3T, SSTM10-2T and SSHM10-5T were distinct from each other and from members of the genus in terms of phenotypic traits related to substrate assimilation. The cell-wall peptidoglycan contained meso-diaminopimelic acid, the major fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0, and the predominant menaquinone was MK-7 for all three strains. Diphosphatidylglycerol, phosphatidylglycerol and an unidentified phospholipid were present in their polar lipid profiles. Based on a polyphasic approach incorporating genomic data, strains SSBR10-3T, SSTM10-2T and SSHM10-5T represent novel species, for which the names Halobacillus salinarum sp. nov. (SSBR10-3T=DSM 114353T=KACC 21935T=NBRC 115504T), Halobacillus shinanisalinarum sp. nov. (SSTM10-2T=DSM 114354T=KACC 21936T=NBRC 115505T) and Halobacillus amylolyticus sp. nov. (SSHM10-5T=DSM 114355T= KACC 21937T=NBRC 115506T) are proposed.

Gracilibacillus salinarum sp. nov. and Gracilibacillus caseinilyticus sp. nov., halotolerant bacteria isolated from a saltern environment.

Two aerobic, Gram-stain-positive, spore-forming motile bacterial strains, designated SSPM10-3T and SSWR10-1T, were isolated from salterns in Jeollanam province of South Korea. Both strains were halotolerant and grew well in 5 % NaCl but not in 20 and 25% NaCl, respectively. Optimal growth was observed with 5 % NaCl, at 30 °C and at pH 7.0-8.0. On the basis of the results of phylogenetic analysis using 16S rRNA gene sequence, both the strains were placed within the genus Gracilibacillus with Gracilibacillus massiliensis (98.65 % similarity) as their nearest neighbour. Menaquinone-7 (MK-7) (97 %) was the major isoprenoid quinone in both strains and major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0. Orthologous average nucleotide identity with usearch (OrthoANIu) and digital DNA-DNA hybridisation (dDDH) percentage comparison indicated that SSPM10-3T and SSWR10-1T exhibited highest similarity with G. massiliensis Awa-1T at 74.27 % and 21.0 and 74.23 % and 20.0 %, respectively. The DNA G+C contents of the strains were 39.1 % (SSPM10-3T) and 38.5 % (SSWR10-1T). Members of the genus Gracilibacillus, both strains were distinct from each other with respect to their ability to produce urease, ß-glucosidase, assimilation of inulin and methyl-α-d-glucopyranoside and degradation of casein. Compared with each other, ANI and d4 dDDH calculations were only 88.2 % and 36.3 %, well below the cut-off values for species delineation for each index. On the basis of their phenotypic, physiological, biochemical and phylogenetic characteristics,SSPM10-3T and SSWR10-1T represent distinct novel species for which names Gracilibacillus salinarum SSPM10-3T and Gracilibacillus caseinilyticus SSWR10-1T are proposed. The type strains are SSPM10-3T (=KACC 21933T =NBRC 115502T) and SSWR10-1T (=KACC 21934T =NBRC 115503T).

Five novel Hymenobacter species isolated from air: Hymenobacter cellulosilyticus sp. nov., Hymenobacter cellulosivorans sp. nov., Hymenobacter aerilatus sp. nov., Hymenobacter sublimis sp. nov. and Hymenobacter volaticus sp. nov.

Five Hymenobacter strains isolated from air samples collected from the Suwon and Jeju regions of the Republic of Korea were studied using polyphasic taxonomic methods. Using 16S rRNA gene sequences and the resulting phylogenetic tree, the strains were primarily identified as members of the genus Hymenobacter. Digital DNA-DNA hybridization values and average nucleotide identities values for species delineation (70 and 95-96 %, respectively) between the five strains and their nearest type strains indicated that each strain represented a novel species. All strains were aerobic, Gram-stain-negative, mesophilic, rod-shaped and catalase- and oxidase-positive, with red to pink coloured colonies. The genome sizes of the five strains varied from 4.8 to 7.1 Mb and their G+C contents were between 54.1 and 59.4 mol%. Based on their phenotypic, chemotaxonomic and genotypic characteristics, we propose to classify these isolates into five novel species within the genus Hymenobacter for which we propose the names, Hymenobacter cellulosilyticus sp. nov., Hymenobacter cellulosivorans sp. nov., Hymenobacter aerilatus sp. nov., Hymenobacter sublimis sp. nov. and Hymenobacter volaticus sp. nov., with strains 5116 S-3T (=KACC 21925T=JCM 35216T), 5116 S-27T (=KACC 21926T=JCM 35217T), 5413 J-13T (=KACC 21928T=JCM 35219T), 5516 S-25T (=KACC 21931T=JCM 35222T) and 5420 S-77T (=KACC 21932T=JCM 35223T) as the type strains, respectively.

Leucobacter allii sp. nov. and Leucobacter rhizosphaerae sp. nov., isolated from rhizospheres of onion and garlic, respectively.

Three bacterial strains, H21R-40T and H21R-36 from garlic (Allium sativum) and H25R-14T from onion (Allium cepa), were isolated from plant rhizospheres sampled in the Republic of Korea. Results of 16S rRNA gene sequence analysis revealed the highest sequence similarity of strain H21R-40T to Leucobacter celer subsp. astrifaciens CBX151T (97.3 %) and Leucobacter triazinivorans JW-1T (97.2 %), and strain H25R-14T to Leucobacter insecticola HDW9BT (98.8 %) and Leucobacter humi Re6T (98.4 %), while the sequence similarity between strains H21R-40T and H21R-36 was 99.8 %. According to the phylogenomic tree, strains H21R-40T with H21R-36 formed an independent clade separable from other Leucobacter species within the genus Leucobacter and strain H25R-14T clustered with Leucobacter insecticola HDW9BT, Leucobacter coleopterorum HDW9AT and Leucobacter viscericola HDW9CT. Strains H21R-40T and H21R-36 had orthologous average nucleotide identity (OrthoANI) and digital DNA-DNA hybridization (dDDH) values (98.1 % and 86.9 %, respectively) higher than the threshold ranges for species delineation (95-96 % and 70 %, respectively). The OrthoANI and dDDH values between two strains (H21R-40T and H25R-14T) and the type strains of species of the genus Leucobacter were lower than 81 and 24 %, respectively. The peptidoglycan type of three strains was type B1. The major menaquinones and major polar lipids of the strains were MK-11 and MK-10, and diphosphatidylglycerol, phatidylglycerol and an unidentified glycolipid, respectively. The major fatty acids (more than 10 % of the total fatty acids) of strains H21R-40T and H21R-36 were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0, and those of strain H25R-14T were anteiso-C15 : 0 and iso-C16 : 0. The phenotypic, chemotaxonomic and genotypic data obtained in this study showed that the strains represent two novel species of the genus Leucobacter, named Leucobacter allii sp. nov. (H21R-40T and H21R-36) and Leucobacter rhizosphaerae sp. nov. (H25R-14T). The respective type strains are H21R-40T (=DSM 114348T=JCM 35241T=KACC 21839T=NBRC 115481T) and H25R-14T (=DSM 114346T=JCM 35239T=KACC 21837T=NBRC 115479T).

Spirosoma oryzicola sp. nov., isolated from dried rice husk.

Strain RHs26T is an aerobic, Gram-stain-negative, non-flagellated and rod- or filamentous-shaped (1.0-1.1×2.3-50 µm) bacterium that was isolated from dried rice husk. It was positive for oxidase and catalase, hydrolysed starch and Tween 80, and weakly hydrolysed CM-cellulose. The strain grew at temperatures between 10 and 37 °C (optimum, 28 °C), in 0-1 % NaCl (optimum, 0 %) and at pH 6.0-9.0 (optimum, pH 7.0-8.0). The predominant membrane fatty acids were summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C16 : 1 ω5c, iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine, an unidentified aminolipid, two unidentified aminophospholipids and two unidentified lipids. The predominant quinone was menaquinone MK-7. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain RHs26T belongs to the genus Spirosoma, presenting the highest sequence similarity to Spirosoma agri S7-3-3T (95.8 %). The genomic DNA G+C content of strain RHs26T was 49.5 %. Strain RHs26T showed the highest orthologous average nucleotide identity (OrthoANI) and digital DNA-DNA hybridization (dDDH) values of 76.4 % and 20.0 % with S. agri KCTC 52727T while sharing OrthoANI and dDDH values of 74.6 % and 19.2 % with Spirosoma terrae KCTC 52035T, the closest relative in the phylogenomic tree. Based on the results of a polyphasic taxonomic study, strain RHs26T represents a novel species in the genus Spirosoma, for which the name Spirosoma oryzicola sp. nov. is proposed. The type strain is RHs26T (=JCM 35224T=KACC 17318T).

Frateuria soli sp. nov. and Frateuria edaphi sp. nov., isolated from greenhouse soil.

Two bacterial strains, designated 5GH9-11T and 5GH9-34T, were isolated from greenhouse soil sampled in Wanju-gun, Jeollabuk-do, Republic of Korea. Both strains formed yellow colonies and were aerobic, rod-shaped and flagellated. The 16S rRNA gene sequence similarity between 5GH9-11T and 5GH9-34T was 98.6 %. Strain 5GH9-11T showed the highest sequence similarities to Dyella thiooxydans ATSB10T (98.1 %) and Frateuria aurantia DSM 6220T (97.7 %) while strain 5GH9-34T revealed the highest sequence similarity to F. aurantia DSM 6220T (98.3 %) and D. thiooxydans ATSB10T (98.3 %). Phylogenetic analysis on the basis of the 16S rRNA gene sequence showed that strains 5GH9-11T and 5GH9-34T formed a robust cluster with Frateuria flava MAH-13T and Frateuria terrea NBRC 104236T. The phylogenomic tree also showed that strains 5GH9-11T and 5GH9-34T formed a robust cluster with F. terrea DSM 26515T and F. flava MAH-13T. Strain 5GH9-11T showed the highest orthologous average nucleotide identity (OrthoANI; 88.5 %) and digital DNA-DNA hybridization (dDDH) values (35.5 %) with F. flava MAH-13T, and strain 5GH9-34T revealed highest OrthoANI (88.1 %) and dDDH (34.2 %) values with F. flava MAH-13T. The orthoANI and dDDH values between strain 5GH9-11T and 5GH9-34T were 87.7 and 33.9 %, respectively. Their major respiratory quinone was ubiquinone 8, and the major cellular fatty acids were iso-C16 : 0, summed feature 9 (iso-C17 : 1 ω9c and/or C16 : 0 10-methyl) and iso-C15 : 0. The major polar lipids of both strains were composed of large or moderate amounts of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminolipid and an unidentified aminophospholipid. Based on these data, strains 5GH9-11T and 5GH9-34T should represent two independent novel species of Frateuria, for which the names Frateuria soli sp. nov. (type strain 5GH9-11T=KACC 16943T=JCM 35197T) and Frateuria edaphi sp. nov. (type strain 5GH9-34T=KACC 16945T=JCM 35198T) are proposed.

Distinct Bacterial and Fungal Communities Colonizing Waste Plastic Films Buried for More Than 20 Years in Four Landfill Sites in Korea.

Plastic pollution has been recognized as a serious environmental problem, and microbial degradation of plastics is a potential, environmentally friendly solution to this. Here, we analyzed and compared microbial communities on waste plastic films (WPFs) buried for long periods at four landfill sites with those in nearby soils to identify microbes with the potential to degrade plastics. Fourier-transform infrared spectroscopy spectra of these WPFs showed that most were polyethylene and had signs of oxidation, such as carbon-carbon double bonds, carbon-oxygen single bonds, or hydrogen-oxygen single bonds, but the presence of carbonyl groups was rare. The species richness and diversity of the bacterial and fungal communities on the films were generally lower than those in nearby soils. Principal coordinate analysis of the bacterial and fungal communities showed that their overall structures were determined by their geographical locations; however, the microbial communities on the films were generally different from those in the soils. For the pulled data from the four landfill sites, the relative abundances of Bradyrhizobiaceae, Pseudarthrobacter, Myxococcales, Sphingomonas, and Spartobacteria were higher on films than in soils at the bacterial genus level. At the species level, operational taxonomic units classified as Bradyrhizobiaceae and Pseudarthrobacter in bacteria and Mortierella in fungi were enriched on the films. PICRUSt analysis showed that the predicted functions related to amino acid and carbohydrate metabolism and xenobiotic degradation were more abundant on films than in soils. These results suggest that specific microbial groups were enriched on the WPFs and may be involved in plastic degradation.

Ferruginibacter albus sp. nov., isolated from a mountain soil, and Mucilaginibacter robiniae sp. nov., isolated from a black locust flower, Robinia pseudoacacia.

Two bacterial strains, designated KIS38-8T and F39-2T, were isolated from a mountain soil sample and a black locust flower (Robinia pseudoacacia) in Republic of Korea, respectively. The phylogenetic tree based on 16S rRNA gene sequences showed that strain KIS38-8T was classified into the genus Ferruginibacter with the highest sequence similarity to Ferruginibacter lapsinanis HU1-HG42T (96.6 %), and strain F39-2T was grouped into the genus Mucilaginibacter with the highest sequence similarity to Mucilaginibacter daejeonensis Jip 10T (97.6 %). Orthologous average nucleotide identity and digital DNA-DNA hybridization values between strain KIS38-8T and closely related Ferruginibacter strains were less than 72 and 19 %, respectively, while those values between strain F39-2T and closely related Mucilaginibacter strains were less than 73 and 21 %, respectively. The DNA G+C contents of strain KIS38-8T and F39-2T were 36.4 and 41.4 mol%, respectively. On the basis of the phenotypic and genotypic evidence, strains KIS38-8T and F39-2T are considered to represent novel species of the genus Ferruginibacter and Mucilaginibacter, respectively, for which the names Ferruginibacter albus sp. nov. (type strain KIS38-8T=KACC 17328T=NBRC 113101T) and Mucilaginibacter robiniae sp. nov. (type strain F39-2T=KACC 19733T=JCM 33062T) have been proposed.

Arachidicoccus terrestris sp. nov., isolated from greenhouse soil.

A Gram-stain-negative, aerobic, non-motile, rod-shaped or occasionally filamentous-shaped bacterial strain, designated 5GH13-10T, was isolated from greenhouse soil sampled in Yeoju-si, Republic of Korea. Colonies were milky-coloured, round and convex, and catalase- and oxidase-positive. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain 5GH13-10T was related to the genus Arachidicoccus and had highest 16S rRNA gene sequence identity with Arachidicoccus rhizosphaerae Vu-144T (98.4 %). The major cellular fatty acids were iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH and summed feature 3 (C16 : 1 ω6c and/or C16 : 1 ω7c). The predominant quinone was menaquinone MK-7, and the polar lipids were composed of phosphatidylethanolamine, one unidentified aminolipid, three unidentified aminophospholipids, one unidentified phospholipid and five unidentified lipids. The genomic DNA G+C content of strain 5GH13-10T was 43.8 mol%. The average nucleotide identity values between strain 5GH13-10T and the closely related species Arachidicoccus ginsenosidivorans Gsoil 809T, Arachidicoccus rhizosphaerae Vu-144T and Archidicoccus soli KIS59-12T were 74.86, 74.74 and 69.52 %, and the digital DNA-DNA hybridization values were 20.0, 19.8 and 18.6 %, respectively. Combined phenotypic, phylogenetic and genomic data demonstrated that strain 5GH13-10T is representative of a novel species of the genus Arachidicoccus, for which we propose the name Arachidicoccus terrestris sp. nov. (type strain 5GH13-10T=KACC 18014T=NBRC 113162T).

Effect of Bacillus mesonae H20-5 Treatment on Rhizospheric Bacterial Community of Tomato Plants under Salinity Stress.

Plant growth-promoting bacteria improve plant growth under abiotic stress conditions. However, their effects on microbial succession in the rhizosphere are poorly understood. In this study, the inoculants of Bacillus mesonae strain H20-5 were administered to tomato plants grown in soils with different salinity levels (EC of 2, 4, and 6 dS/m). The bacterial communities in the bulk and rhizosphere soils were examined 14 days after H20-5 treatment using Illumina MiSeq sequencing of the bacterial 16S rRNA gene. Although the abundance of H20-5 rapidly decreased in the bulk and rhizosphere soils, a shift in the bacterial community was observed following H20-5 treatment. The variation in bacterial communities due to H20-5 treatment was higher in the rhizosphere than in the bulk soils. Additionally, the bacterial species richness and diversity were greater in the H20-5 treated rhizosphere than in the control. The composition and structure of the bacterial communities varied with soil salinity levels, and those in the H20-5 treated rhizosphere soil were clustered. The members of Actinobacteria genera, including Kineosporia, Virgisporangium, Actinoplanes, Gaiella, Blastococcus, and Solirubrobacter, were enriched in the H20-5 treated rhizosphere soils. The microbial co-occurrence network of the bacterial community in the H20-5 treated rhizosphere soils had more modules and keystone taxa compared to the control. These findings revealed that the strain H20-5 induced systemic tolerance in tomato plants and influenced the diversity, composition, structure, and network of bacterial communities. The bacterial community in the H20-5 treated rhizosphere soils also appeared to be relatively stable to soil salinity changes.

Protaetiibacter larvae sp. nov. and Agromyces intestinalis sp. nov., isolated from the gut of larvae of Protaetia brevitarsis seulensis, reclassification of Lysinimonas yzui as Pseudolysinimonas yzui comb. nov. and emended description of the genus Pseudolysinimonas.

Two bacterial strains, FWR-8T and CFWR-9T, were isolated from the gut of larvae of Protaetia brevitarsis seulensis that were raised at the National Institute of Agricultural Sciences, Wanju-gun, Republic of Korea. Both strains were strictly aerobic, Gram-stain-positive and non-motile. Strain FWR-8T possessed the highest sequence similarity (98.7 %) to that of Protaetiibacter intestinalis 2DFWR-13T and the phylogenetic tree revealed that strain FWR-8T formed a cluster with Ptb. intestinalis 2DFWR-13T. Pseudolysinimonas kribbensis MSL-13T and Lysinimonas yzui N7XX-4T shared a high 16S rRNA gene sequence similarity (97.8 %) and formed a cluster adjacent to the cluster that included Ptb. intestinalis 2DFWR-13T. The 16S rRNA gene sequence of strain CFWR-9T exhibited the highest similarity (97.7 %) to that of Agromyces binzhouensis OAct353T and the phylogenetic tree indicated that strain CFWR-9T formed one independent cluster with A. binzhouensis OAct353T that was within the radius of the genus Agromyces. The peptidoglycan type, major fatty acids, major menaquinones and total polar lipids of strain FWR-8T were characterized as type B1, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0, MK-15, MK-16 and MK-14, and diphosphatidylglycerol, phosphatidylglycerol, two unidentified glycolipids and one unidentified lipid, respectively. Those from strain CFWR-9T were type B1, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0, MK-11, MK-12 and MK-10, and diphosphatidylglycerol, phosphatidylglycerol, two unidentified glycolipids and one unidentified lipid, respectively. Based on the phenotypic and genotypic data, strains FWR-8T and CFWR-9T each represent a novel species within the genera Protaetiibacter and Agromyces, respectively. For these species, the names Protaetiibacter larvae sp. nov. and Agromyces intestinalis sp. nov. have been proposed, with the type strains FWR-8T (=KACC 19322T=NBRC 113051T) and CFWR-9T (=KACC 19306T=NBRC 113046T), respectively. Our results also justify a reclassification of Lysinimonas yzui as Pseudolysinimonas yzui comb. nov. and an emended description of the genus Pseudolysinimonas isprovided.

Arachidicoccus soli sp. nov., a bacterium isolated from soil.

A Gram-stain-negative, aerobic, non-motile and rod-shaped bacterium, designated KIS59-12T, was isolated from a soil sample collected on Hodo island, Boryeong, Republic of Korea. The strain grew at 10-33 °C, pH 6.0-7.5 and with 0-4 % NaCl (w/v). Results of phylogenetic analysis based on 16S rRNA gene sequences showed that strain KIS59-12T was in the same clade as Arachidicoccus rhizosphaerae Vu-144T and Arachidicoccus ginsenosidivorans Gsoil809T with 97.5 and 97.2 % sequence similarity, respectively. Comparative genome analysis between strain KIS59-12T and A. rhizosphaerae Vu-144T showed that average nucleotide identity value was 69.4 % and the digital DNA-DNA hybridization value was 19.1 %. The major respiratory quinone was menaquinone-7. The major polar lipids were phosphatidylethanolamine and an unknown polar lipid. The predominant cellular fatty acids were iso-C15 : 0, iso-C15 : 1 G and iso-C17 : 0 3-OH, which supported the affiliation of strain KIS59-12T with the genus Arachidicoccus. The major polyamines were homospermidine and putrescine. The genomic DNA G+C content was 36.4 mol%. On the basis of phylogenetic, physiological and chemotaxonomic characteristics, strain KIS59-12T represents a novel species of the genus Arachidicoccus, for which the name Arachidicoccus soli sp. nov. is proposed. The type strain of Arachidicoccus soli is KIS59-12T (=KACC 17340T=NBRC 113161T).

Chitinophaga agri sp. nov., a bacterium isolated from soil of reclaimed land.

A Gram-negative, aerobic, and long rod-shaped bacterium, designated as H33E-04T, was isolated from the soil of reclaimed land, Republic of Korea. The strain grew at a temperature range of 15-40 °C, pH 5.0-10.0, and 0-2% NaCl (w/v). The phylogenetic analysis based on 16S rRNA gene sequences showed that strain H33E-04T was in the same clade with Chitinophaga pinensis DSM 2588T, Chitinophaga filiformis IFO 15056T, and Chitinophaga ginsengisoli Gsoil 052T with 98.4%, 97.9%, and 97.8% sequence similarities, respectively. The de novo genome assembly revealed that the DNA G + C content of the strain was 46.2 mol%. Comparative genome analysis between strain H33E-04T and C. pinensis DSM 2588 T showed that the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values were 79.9% and 23.4%, respectively. The major respiratory quinone was menaquinone-7 (MK-7) and the predominant cellular fatty acids were iso-C15:0 (31.7%), C16:1 ω5c (31.2%), and iso-C17:0 3-OH (11.8%), supporting the affiliation of strain H33E-04T with the genus Chitinophaga. Based on phylogenetic, physiological, and chemotaxonomic characteristics, strain H33E-04T represents a novel species of the genus Chitinophaga, for which the name Chitinophaga agri sp. nov. is proposed. The type strain of Chitinophaga agri is H33E-04T (= KACC 21303T = NBRC114512T).

Different types of agricultural land use drive distinct soil bacterial communities.

Biogeographic patterns in soil bacterial communities and their responses to environmental variables are well established, yet little is known about how different types of agricultural land use affect bacterial communities at large spatial scales. We report the variation in bacterial community structures in greenhouse, orchard, paddy, and upland soils collected from 853 sites across the Republic of Korea using 16S rRNA gene pyrosequencing analysis. Bacterial diversities and community structures were significantly differentiated by agricultural land-use types. Paddy soils, which are intentionally flooded for several months during rice cultivation, had the highest bacterial richness and diversity, with low community variation. Soil chemical properties were dependent on agricultural management practices and correlated with variation in bacterial communities in different types of agricultural land use, while the effects of spatial components were little. Firmicutes, Chloroflexi, and Acidobacteria were enriched in greenhouse, paddy, and orchard soils, respectively. Members of these bacterial phyla are indicator taxa that are relatively abundant in specific agricultural land-use types. A relatively large number of taxa were associated with the microbial network of paddy soils with multiple modules, while the microbial network of orchard and upland soils had fewer taxa with close mutual interactions. These results suggest that anthropogenic agricultural management can create soil disturbances that determine bacterial community structures, specific bacterial taxa, and their relationships with soil chemical parameters. These quantitative changes can be used as potential biological indicators for monitoring the impact of agricultural management on the soil environment.

Paenibacillus lycopersici sp. nov. and Paenibacillus rhizovicinus sp. nov., isolated from the rhizosphere of tomato (Solanum lycopersicum).

Two Gram-stain-positive, rod-shaped, endospore-forming bacteria, designated 12200R-189T and 14171R-81T were isolated from the rhizosphere of tomato plants. The 16S rRNA gene sequence similarity between strains 12200R-189T and 14171R-81T were 97.2%. Both strains showed the highest 16S rRNA gene sequence similarities to Paenibacillus sacheonensis SY01T (96.3% and 98.0%, respectively). The genome of strain 12200R-189T was approximately 6.7 Mb in size with 5,750 protein-coding genes (CDSs) and the G + C content was 58.1 mol%, whereas that of strain 14171R-81T comprised one chromosome of 7.0 Mb and two plasmids (0.2 Mb each) with 6,595 CDSs and the G + C content was 54.5 mol%. Comparative genome analysis revealed that average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values among 12200R-189T, 14171R-81T, and other closely related species were below the cut-off levels 95% and 70%, respectively. Strain 12200R-189T grew at a temperature range of 15-40°C, pH 6.0-9.0, and 0-3% NaCl (w/v), whereas strain 14171R-81T grew at a temperature range of 10-37°C, pH 6.0-8.0, and 0-1% NaCl (w/v). Menaquinone-7 (MK-7) was the only isoprenoid quinone detected in both strains. The predominant cellular fatty acids (> 10%) were iso-C15:0, anteiso-C15:0, and iso-C16:0. The polar lipids of strain 12200R-189T were diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), aminophospholipid (APL), phospholipid (PL), phosphatidylglycolipid (PGL), and four aminophosphoglycolipids (APGLs) and those of strain 14171R-81T were DPG, PG, PE, APL, three PLs, two PGLs, and three APGLs. Based on phylogenetic, genomic, phenotypic, and chemotaxonomic analyses, strains 12200R-189T and 14171R-81T represent two novel species of the genus Paenibacillus, for which the names Paenibacillus lycopersici sp. nov. and Paenibacillus rhizovicinus sp. nov. are proposed. The type strains are 12200R-189T (= KACC 19916T = CCTCC AB 2020027T) and 14171R-81T (= KACC 19915T = CCTCC AB 2020026T).

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).

Xylophilus rhododendri sp. nov., Isolated from Flower of Royal Azalea, Rhododendron schlippenbachii.

A bacterial strain, designated CJ1-R5T, was isolated from the flower of the royal azalea plant (Rhododendron schlippenbachii) collected in Jeju Island, Republic of Korea. The strain was a Gram-negative, strictly aerobic, motile, rod-shaped bacterium, growing at a temperature range of 4-33 °C (optimum 28-30 °C), pH 5.0-9.0 (optimum pH 7.0-8.0), and 0-1% NaCl (optimum 0%). The 16S rRNA sequence analysis of strain CJ1-R5T revealed the highest sequence similarity (97.9%) with Xylophilus ampelinus ATCC 33914T, and sequence similarities of less than 97.2% with other validly named species. Phylogenetic tree analysis based on the 16S rRNA gene sequences showed that strain CJ1-R5T clustered with Xylophilus ampelinus ATCC 33914T and two uncultured bacterial clones. The only quinone observed in strain CJ1-R5T was ubiquinone-8. The polar lipids observed were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified aminophospholipid and two unidentified lipids. The major fatty acids were C16:0, C17:0 cyclo, and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The genome size of strain CJ1-R5T was 5.85 Mbp. The genomic G + C content was 68.4 mol%. ANI and dDDH values between strain CJ1-R5T and Xylophilus ampelinus ATCC 33914T were 79.0% and 22.5%, respectively. Based on the polyphasic taxonomic data, strain CJ1-R5T is considered to represent a novel species, for which the name Xylophilus rhododendri sp. nov. is proposed. The type strain is CJ1-R5T (= KACC 21265T = CCTCC AB2020030T).

Alteration of Bacterial Wilt Resistance in Tomato Plant by Microbiota Transplant.

Plant-associated microbiota plays an important role in plant disease resistance. Bacterial wilt resistance of tomato is a function of the quantitative trait of tomato plants; however, the mechanism underlying quantitative resistance is unexplored. In this study, we hypothesized that rhizosphere microbiota affects the resistance of tomato plants against soil-borne bacterial wilt caused by Ralstonia solanacearum. This hypothesis was tested using a tomato cultivar grown in a defined soil with various microbiota transplants. The bacterial wilt-resistant Hawaii 7996 tomato cultivar exhibited marked suppression and induction of disease severity after treatment with upland soil-derived and forest soil-derived microbiotas, respectively, whereas the transplants did not affect the disease severity in the susceptible tomato cultivar Moneymaker. The differential resistance of Hawaii 7996 to bacterial wilt was abolished by diluted or heat-killed microbiota transplantation. Microbial community analysis revealed the transplant-specific distinct community structure in the tomato rhizosphere and the significant enrichment of specific microbial operational taxonomic units (OTUs) in the rhizosphere of the upland soil microbiota-treated Hawaii 7996. These results suggest that the specific transplanted microbiota alters the bacterial wilt resistance in the resistant cultivar potentially through a priority effect.

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).

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).