- Volume 66, Issue 4, 2016

The actinomycete strain N74T, isolated from cave soil, was studied using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain N74T formed a stable, distinct lineage cluster together with Microbispora thailandensis NN276T (99.3 % similarity) and Microbispora mesophila JCM 3151T (97.5 %). Strain N74T was observed to produce single spherical spores on aerial mycelium as reported for M. mesophila JCM 3151T and M. thailandensis NN276T but different from other known species of the genus Microbispora, which are characterized by pairs of spores on aerial hyphae. Multilocus sequence analyses based on concatenated partial gyrB, rpoB, atpD, recA and 16S rRNA gene sequences showed a clear distinction of strain N74T, M. mesophila JCM 3151T and M. thailandensis NN276T from other members of the genus Microbispora, although the chemotaxonomic characteristics of strain N74T were similar to the genus Microbispora; the cell wall contained meso-diaminopimelic acid and the whole-cell hydrolysate contained madurose as the diagnostic sugar. The major menaquinone was MK-9(H4). The fatty acid profile contained iso-C16 : 0. On the basis of morphological, chemotaxonomic and phylogenetic evidence, strain N74T is assigned to a novel species of a new genus, for which the name Sphaerimonospora cavernae gen. nov., sp. nov. is proposed. The type strain of Sphaerimonospora cavernae is N74T ( = BCC 77604T = NBRC 111481T). It is also proposed that M. mesophila and M. thailandensis be transferred to this genus as Sphaerimonospora mesophila comb. nov. (type strain JCM 3151T = NBRC 14179T = DSM 43048T) and Sphaerimonospora thailandensis comb. nov. (type strain NN276T = BCC 41490T = NBRC 107569T), respectively.

A Gram-stain-positive, aerobic and non-motile actinobacterial strain, designated YIM C0533T, was isolated from a soil sample collected from Shiling county, Yunnan province, south-west China. The isolate grew at 15–37 °C, pH 6.0–8.0 and in the presence of 0–3 % (w/v) NaCl. The whole-cell hydrolysates contained meso-diaminopimelic acid, xylose, galactose, mannose, ribose, arabinose, glucose and rhamnose. The acyl type of muramic acid was glycolyl. The polar lipids detected were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and an unidentified phospholipid. The major cellular fatty acids were iso-C16 : 0, 10-methyl C17 : 0 and iso-C15 : 0.MK-9(H6) was the predominant menaquinone. The genomic DNA G+C content was determined to be 69.4 mol%. These chemotaxonomic data and the morphological properties were consistent with those of the genus Hamadaea. The strain showed highest sequence similarities to Hamadaea tsunoensis CGMCC 4.1403T on phylogenetic analysis of 16S rRNA gene sequences and was found to form a coherent cluster in the neighbour–joining tree. The DNA–DNA hybridization experiment indicated that the DNA–DNA relatedness value between strain YIM C0533T and H. tsunoensis CGMCC 4.1403T was 34.4 ± 1.3 %. In addition, the results of physiological and biochemical tests allowed the isolate to be differentiated phenotypically from H. tsunoensis CGMCC 4.1403T. On the basis of data from this polyphasic study, strain YIM C0533T is characterized as a novel species of the genus Hamadaea, for which the name Hamadaea flava sp. nov. is proposed. The type strain is YIM C0533T ( = CPCC 204160T = KCTC 39591T = CGMCC 4.7289T). An emended description of the genus Hamadaea is also provided.

A novel actinobacterium designated strain MWE-A11T was isolated from the root of wild Artemisia princeps (mugwort). The isolate was aerobic, Gram-stain-positive and short rod-shaped, and the colonies were yellow and circular with entire margin. Strain MWE-A11T grew at 15–37 °C and pH 6.0–8.0. The predominant isoprenoid quinones were MK-11 and MK-10. The predominant fatty acids were anteiso-C15 : 0 and iso-C16 : 0, and the DNA G+C content was 68.8 mol%. The main polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. The peptidoglycan contained 2,4-diaminobutyric acid as the diagnostic diamino acid, and the acyl type was glycolyl. Phylogenetic analyses based on 16S rRNA gene sequence comparisons indicated that strain MWE-A11T was affiliated with the family Microbacteriaceae, and was most closely related to the type strains of Humibacter antri (96.4 % 16S rRNA gene sequence similarity), Herbiconiux moechotypicola (96.3 %), Leifsonia soli (96.3 %), Leifsonia lichenia (96.2 %), Leifsonia xyli subsp. cynodontis (96.1 %), Microbacterium testaceum (96.0 %) and Humibacter albus (96.0 %). However, the combination of chemotaxonomic properties clearly distinguished strain MWE-A11T from the related taxa at genus level. Accordingly, Allohumibacter endophyticus gen. nov., sp. nov. is proposed to accommodate a new member of the family Microbacteriaceae. The type strain of the type species is MWE-A11T ( = JCM 19371T = KCTC 29232T).

A Gram-stain-positive bacterium, designated T14T, was isolated from the gut of Holotrichia oblita larvae and was subjected to a taxonomic study. The isolate was rod-shaped, aerobic, non-motile, non-spore-forming and yellow-pigmented. Phylogenetic analysis based on 16S rRNA gene sequence comparison indicated that the isolate is related to the genus Leucobacter. Its closest neighbours were the type strains ‘Leucobacter kyeonggiensis’ F3-P9 (96.8 % 16S rRNA gene sequence similarity), Leucobacter celer NAL101T (96.2 %) and Leucobacter chironomi DSM 19883T (95.5 %). The DNA G+C content of strain T14T was 69.3 mol%, and DNA–DNA hybridization values with closely related strains were < 32 %. The predominant cellular fatty acids were anteiso-C15 : 0 (49.3 %), iso-C16 : 0 (16.4 %) and anteiso-C17 : 0 (16.8 %). The major polar lipids were aminolipid, diphosphatidylglycerol, phosphatidylglycerol, phospholipid, phosphoglycolipid and unidentified glycolipids. The predominant respiratory quinone was MK-11. Based on these phylogenetic and phenotypic results, strain T14T can be clearly distinguished from all of the recognized species of the genus Leucobacter and is considered to represent a novel species of the genus Leucobacter. The name Leucobacter holotrichiae sp. nov. is proposed, with the type strain T14T ( = DSM 28968T = JCM 30245T).

Strain RP1T, a Gram-stain-positive, non-motile, non-spore-forming, coccus-shaped bacterium, was isolated from drainage of India Pesticides Limited, a lindane-producing unit situated at Chinhat, Lucknow, India. 16S rRNA gene sequence analysis revealed that strain RP1T belongs to the family Propionibacteriaceae and was closely related to the members of the genus Tessaracoccus with a similarity range of 95.4–97.6 %. Strain RP1T was facultatively anaerobic, oxidase-negative, catalase-positive and capable of nitrate reduction. Strain RP1T contained peptidoglycan type A3γ′, with ll-diaminopimelic acid as the diagnostic diamino acid and glycine at position 1 of the peptide subunit. The major cellular fatty acid of strain RP1T was anteiso-C15 : 0 but a significant amount of iso-C14 : 0 was also detected. MK-9(H4) was the major respiratory quinone and polyamines detected were spermine and spermidine. The polar lipids included diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids and two unknown phospholipids. The G+C content of the DNA was 66.7 mol%. The levels of DNA–DNA relatedness between RP1T and Tessaracoccus lubricantis KSS-17SeT, Tessaracoccus oleiagri SL014B-20A1T and Tessaracoccus flavescens SST-39T were 49.8, 34.8 and 23.5 %, respectively. Based on the phenotypic and phylogenetic data presented, strain RP1T can be differentiated from previously described species of the genus Tessaracoccus, and thus represents a novel species, for which the name Tessaracoccus flavus sp. nov. is proposed. The type strain is RP1T ( = DSM 100159T = MCC 2769T = KCTC 39686T).

A Gram-stain-positive, aerobic organism was isolated as an endophyte from the stem tissue of healthy maize (Zea mays) and investigated in detail for its taxonomic position. On the basis of the 16S rRNA gene sequence analysis, strain JM-601T was shown to be most closely related to Nocardioides alpinus (98.3 %), and Nocardioides ganghwensis (98.0 %). The 16S rRNA gene sequence similarity to all other species of the genus Nocardioides was ≤ 98.0 %. The diagnostic diamino acid of the peptidoglycan was ll-diaminopimelic acid. The major quinone of strain JM-601T was menaquinone MK-8(H4). The polar lipid profile revealed the major components diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylcholine and an unidentified phospholipid. The polyamine pattern contained predominantly spermine and moderate amounts of spermidine. In the fatty acid profile, iso-C16 : 0, C17 : 1ω8c and 10-methyl C17 : 0 were present in major amounts. All these data support the allocation of the strain to the genus Nocardioides. The results of physiological and biochemical characterization allow in addition a phenotypic differentiation of strain JM-601T from N. alpinus and N. ganghwensis. Strain JM-601T represents a novel species of the genus Nocardioides, for which we propose the name Nocardioides zeicaulis sp. nov., with JM-601T ( = CCM 8654T = CIP 110980T) as the type strain.

A novel actinobacterial strain, AD1-86T, was isolated from the vaginal fluid of a Korean female and was characterized by a polyphasic approach. The strain was a facultatively anaerobic, Gram-stain-positive, non-spore-forming, non-motile, catalase-positive and oxidase-negative short rod. Colonies were creamy white, of low convexity and 1–2 mm in diameter after growth on DSM 92 agar plates at 37 °C for 2 days. The most closely related strains were Dermabacter hominis DSM 7083T and Helcobacillus massiliensis 6401990T (98.3 and 96.3 % 16S rRNA gene sequence similarity, respectively). The isolate grew optimally at 37 °C and pH 7 in the presence of 0.5 % (w/v) NaCl. The cell-wall peptidoglycan contained meso-diaminopimelic acid and the cell-wall hydrolysates contained ribose, galactose and glucose. The DNA G+C content was 62.6 mol% and the mean DNA–DNA relatedness value of the isolate to D. hominis DSM 7083T was 31.1 ± 3.0 % (reciprocal: 48.2 ± 5.3 %). The major cellular fatty acids (>10 %) were anteiso-C17 : 0, anteiso-C15 : 0 and iso-C16 : 0, and the menaquinones were MK-9, MK-8 and MK-7. The polar lipid profile of strain AD1-86T consisted of diphosphatidylglycerol, phosphatidylglycerol, two aminolipids and a glycolipid. Data from this polyphasic study indicate that strain AD1-86T represents a novel species of the genus Dermabacter, for which the name Dermabacter vaginalis sp. nov. is proposed; the type strain is AD1-86T ( = KCTC 39585T = DSM 100050T).

A novel strain, designated AM23T, was isolated from the gut of a purple sea urchin Heliocidaris crassispina collected from the coastal waters of the Korean island Dokdo. 16S rRNA gene sequence analysis showed that strain AM23T belonged to the genus Arthrobacter in the family Micrococcaceae and shared highest sequence similarity with Arthrobacter agilis DSM 20550T (98.77 %). Strain AM23T was catalase-positive, oxidase-negative and grew optimally at 20 °C, in the presence of 1 % (w/v) NaCl and at pH 7. The isolate was a Gram-stain-positive, non-motile, strictly aerobic and coccus-shaped bacterium. The major cellular fatty acids were anteiso-C15 : 0 and iso-C15 : 0. The polar lipids of strain AM23T were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, one unidentified glycolipid and four unidentified lipids. The components of the cell-wall peptidoglycan were lysine, glutamic acid and alanine and the predominant cell-wall sugars were galactose, mannose, rhamnose and ribose. The major respiratory quinone was identified as menaquinone MK-9(H2). The genomic DNA G+C content was 67.3 mol% and the DNA–DNA hybridization values showed the strain shared less than 29 % genomic relatedness with A. agilis DSM 20550T. The results of the phylogenetic, phenotypic and genotypic analysis indicate that strain AM23T represents a novel species in the genus Arthrobacter, for which the name Arthrobacter echini sp. nov. is proposed. The type strain is AM23T ( = KACC 18260T = DSM 29493T).

A halotolerant actinobacterial strain, designated EGI 80423T, was isolated from a desert soil of Xinjiang, north-west China, and subjected to a polyphasic taxonomic characterization. Strain EGI 80423T grew at pH 7.0–10.0 and with 0–14.0 % (w/v) NaCl, optimally at pH 8.0–9.0 and with 2.0–4.0 % (w/v) NaCl. Cells of strain EGI 80423T were Gram-stain-positive, non-motile cocci with diameters of 0.6–0.8 μm. The diagnostic diamino acid of the peptidoglycan was ornithine, and the interpeptide bridge was Orn ← Glu. The major fatty acids identified were iso-C17 : 1ω9c, iso-C15 : 0 and iso-C17 : 0. The predominant menaquinone was MK-8(H4), while the polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown phospholipids, two unknown glycolipids, six unknown phosphoglycolipids and five unknown polar lipids. The G+C content of the genomic DNA was 72.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EGI 80423T clustered with the single member of the genus Ornithinicoccus. Sequence similarity between strain EGI 80423T and Ornithinicoccus hortensis NBRC 16434T. Because the type strain has been provided by NBRC, Japan was 97.7 %. The DNA–DNA relatedness value between strain EGI 80423T and O. hortensis NBRC 16434T was 36.84 %. Based on morphological, chemotaxonomic and phylogenetic characteristics, and DNA–DNA hybridization data, strain EGI 80423T represents a novel species of the genus Ornithinicoccus, for which the name Ornithinicoccus halotolerans sp. nov. is proposed. The type strain is EGI 80423T ( = CGMCC 1.14989T = KCTC 39700T). The description of the genus Ornithinicoccus has also been emended.

A novel actinomycete, designated strain 1H-HV4T, was isolated from the head of Camponotus japonicas Mayr, which was collected from Northeast Agriculture University (Harbin, Heilongjiang, China). Chemotaxonomic properties of this strain were consistent with those of members of the genus Nocardia. The cell wall contained meso-diaminopimelic acid and whole-cell sugars were galactose, glucose and arabinose. The predominant menaquinone was MK-8(H4,ω-cycl). The phospholipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were identified as C18 : 0 10-methyl, C16 : 0, C18 : 1ω9c and C16 : 1ω7c. Mycolic acids were found to be present. 16S rRNA gene sequence analysis also showed that strain 1H-HV4T was a member of the genus Nocardia, with the highest sequence similarities to Nocardia salmonicida JCM 4826T (97.39 %), Nocardia soli JCM 11441T (97.12 %) and Nocardia cummidelens JCM 11439T (97.08 %). 16S rRNA gene sequence similarities to type strains of other members of the genus Nocardia were less than 97 %. However, DNA–DNA relatedness values and phenotypic data demonstrated that strain1H-HV4T was clearly distinguished from all closely related species of the genus Nocardia. It is concluded that the isolate can be classified as representing a novel species of the genus Nocardia, for which the name Nocardia camponoti is proposed. The type strain is 1H-HV4T ( = DSM 100526T = CGMCC 4.7278T).

A novel bacterial strain, CJ22T, was isolated from soil of a ginseng field located in Anseong, Korea. Cells of strain CJ22T were aerobic, Gram-stain-positive, endospore-forming, motile, oxidase- and catalase-positive and rod-shaped. The isolate grew optimally at pH 7 and 30 °C. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CJ22T belonged to the genus Cohnella, displaying highest sequence similarity of 97.3 % with Cohnella panacarvi Gsoil 349T. DNA–DNA relatedness between strain CJ22T and its closest relative was 35.5 % (reciprocal value, 23.8 %). The phenotypic features of strain CJ22T also distinguished it from related species of the genus Cohnella. The diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. The major isoprenoid quinone was menaquinone MK-7 and the major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, lysyl-phosphatidylglycerol, two unidentified phospholipids and two unidentified aminophospholipids. The predominant cellular fatty acids of strain CJ22T were anteiso-C15 : 0, iso-C16 : 0 and C16 : 0. The DNA G+C content was 63.1 mol%. Based on data from this polyphasic taxonomic study, strain CJ22T is considered to represent a novel species of the genus Cohnella, for which the name Cohnella saccharovorans sp. nov. is proposed. The type strain is CJ22T ( = KACC 17501T = JCM 19227T).

A strictly anaerobic, mesophilic, syntrophic, alkane-degrading strain, L81T, was isolated from a biofilm sampled from a black smoker chimney at the Loki's Castle vent field. Cells were straight, rod-shaped, Gram-positive-staining and motile. Growth was observed at pH 6.2–9.5, 14–42 °C and 0.5–6 % (w/w) NaCl, with optima at pH 7.0–8.2, 37 °C and 3 % (w/w) NaCl. Proteinaceous substrates, sugars, organic acids and hydrocarbons were utilized for growth. Thiosulfate was used as an external electron acceptor during growth on crude oil. Strain L81T was capable of syntrophic hydrocarbon degradation when co-cultured with a methanogenic archaeon, designated strain LG6, isolated from the same enrichment. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain L81T is affiliated with the family Lachnospiraceae, and is most closely related to the type strains of Natranaerovirga pectinivora (92 % sequence similarity) and Natranaerovirga hydrolytica (90 %). The major cellular fatty acids of strain L81T were C15 : 0, anteiso-C15 : 0 and C16 : 0, and the profile was distinct from those of the species of the genus Natranaerovirga. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, three unidentified phospholipids, four unidentified glycolipids and two unidentified phosphoglycolipids. The G+C content of genomic DNA was determined to be 31.7 mol%. Based on our phenotypic, phylogenetic and chemotaxonomic results, strain L81T is considered to represent a novel species of a new genus of the family Lachnospiraceae, for which we propose the name Abyssivirga alkaniphila gen. nov., sp. nov. The type strain of Abyssivirga alkaniphila is L81T ( = DSM 29592T = JCM 30920T). We also provide emended descriptions of Natranaerovirga pectinivora and Natranaerovirga hydrolytica.

Creeping thistle [Cirsium arvense (L.) Scop.] and dahlia (Dahlia sp.) plants showing typical symptoms of phytoplasma infection including yellowing, stunting, inflorescence and proliferation, were sampled; the presence of phytoplasma was confirmed by standard PCR using universal primers. RFLP analysis allowed classification of the detected phytoplasma strains CirYS, CirYS1 and DahlP within the 16SrXI group, the unique restriction profile F2nR2 fragment obtained in silico by iPhyClassifier indicated that they belong to the new 16SrXI-E subgroup. Genetic analysis of the 16S rRNA gene revealed that the studied strains shared less than 97.5 % similarity with all of the previously described ‘Candidatus Phytoplasma’ species. The closest relatives are ‘Candidatus Phytoplasma cynodontis’ and ‘Candidatus Phytoplasma oryzae’ with 96.8 % and 96.6 % similarity. All strains studied bear three specific regions in the 16S rRNA gene, discriminating them from the other phytoplasma species. Phylogenetic analysis of the 16S rRNA and secA genes confirmed this specificity, as the creeping thistle and dahlia phytoplasma strains clustered in a distinguishable lineage group. The uniqueness of the genetic analysis agrees with the biological characterization of the studied phytoplasma strains, their host range, and geographical distribution. The strains only infect dicotyledonous plants in Europe, contrary to their closest relatives. Based on their unique properties, it could be concluded that the studied phytoplasma strains represent a discrete group that is proposed as a novel taxon ‘Candidatus Phytoplasma cirsii’, with strain CirYS as a reference strain.

A Gram-reaction-positive and rod-shaped bacterium, designated KN3-8-4T, was isolated from shrimp paste collected from a market in Nakhon Sri Thammarat province, Thailand. Strain KN3-8-4T was a strictly aerobic, motile bacterium that produced ellipsoidal endospores at a terminal position. The isolate grew at pH 4.5–10 (optimum pH 7.5), at 11–42 °C (optimum 37 °C) and in the presence of 0–15 % (w/v) NaCl (optimum 1–5 %). On the basis of 16S rRNA gene sequences, strain KN3-8-4T belonged to the genus Virgibacillus and showed highest similarity with Virgibacillus olivae JCM 30551T (97.85 % 16S rRNA gene sequence similarity). The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid and the major isoprenoid quinone was MK-7. Strain KN3-8-4T contained anteiso-C15 : 0 and anteiso-C17 : 0 as major cellular fatty acids and had phosphatidylglycerol, diphosphatidylglycerol, two unknown phospholipids and one glycolipid as polar lipids. The DNA G+C content was 43.5 mol%. Strain KN3-8-4T showed low DNA–DNA relatedness (20.44 %) with V. olivae JCM 30551T. On the basis of phenotypic and chemotaxonomic data and phylogenetic analyses, strain KN3-8-4T represents a novel species of the genus Virgibacillus for which the name Virgibacillus kapii sp. nov. is proposed. The type strain is KN3-8-4T ( = JCM 30071T = LMG 28282T = TISTR 2279T = PCU 345T).

A bacterial strain designated PM10T was isolated from root nodules of Periandra mediterranea in Brazil. Phylogenetic analyses based on 16S rRNA gene sequences placed the isolate in the genus Paenibacillus with its closest relatives being Paenibacillus vulneris CCUG 53270T and Paenibacillus yunnanensis YN2T with 95.6 and 95.9 % 16S rRNA gene sequence similarity, respectively. The isolate was a Gram-stain-variable, motile, sporulating rod that was catalase-negative and oxidase-positive. Caseinase was positive, amylase was weakly positive and gelatinase was negative. Growth was supported by many carbohydrates and organic acids as carbon sources. MK-7 was the only menaquinone detected and anteiso-C15 : 0 was the major fatty acid. Major polar lipids were diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol and two unidentified lipids. meso-Diaminopimelic acid was detected in the peptidoglycan. The DNA G+C content was 52.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain PM10T should be considered representative of a novel species of the genus Paenibacillus, for which the name Paenibacillus periandrae sp. nov. is proposed. The type strain is PM10T ( = LMG 28691T = CECT 8827T).

A Gram-stain-negative, obligately aerobic, non-motile, non-sporulating, rod-shaped bacterium, designated TZCO2T, was isolated from the soil of an irrigated coffee plantation in Arusha, Tanzania, East Africa. Phylogenetic analysis, based on 16S rRNA gene sequences, indicated that the isolate is affiliated with the genus Taibaiella in the family Chitinophagaceae. Its closest relative is Taibaiella koreensis THG-DT86T (96.7 %). The pH and temperature ranges for growth were pH 6.0–8.5 (optimum 7.0–7.5) and 10–35 °C (optimum 30 °C, respectively. The predominant fatty acids were iso-C15 : 0 (32.4 %), iso-C15 : 1 G (22.6 %), iso-C17 : 0 (15.1 %) and iso-C17 : 0 3-OH (10.0 %) The only isoprenoid quinone detected in strain TZCO2T was menaquinone-7 (MK-7); the major polar lipids were phosphoaminolipid, phosphatidylethanolamine, unidentified aminolipids and lipids. The DNA G+C content was 51.9 mol%. Physiological and chemotaxonomic data further confirmed that strain TZCO2T is distinct from other members of the genus Taibaiella. Thus, strain TZCO2T is considered to represent a novel species of the genus, for which the name Taibaiella coffeisoli sp. nov. is proposed. The type strain is TZCO2T ( = NCAIM B 02601T = CCM 8601T).

A bacterial strain, designated shin9-1T, was isolated from a water sample taken from a water convolvulus field in Taiwan and characterized using a polyphasic taxonomical approach. Cells of strain shin9-1T were aerobic, Gram-stain-negative, rod-shaped and surrounded by a thick capsule and formed cream-coloured colonies. Growth occurred at 10–45 °C (optimum, 30 °C), with 0–3.0 % NaCl (optimum, 0.5 %) and at pH 7.0–9.0 (optimum, pH 7.0). Strain shin9-1T did not form nodules on a legume plant, Macroptilium atropurpureum, and the nodulation genes nodA, nodC and the nitrogenase reductase gene nifH were not detected by PCR. Phylogenetic analyses based on 16S rRNA and three housekeeping gene sequences (recA, atpD and rpoB) showed that strain shin9-1T belonged to the genus Rhizobium. Strain shin9-1T had the highest level of 16S rRNA gene sequence similarity with respect to Rhizobium daejeonense L61T (97.6 %). The major fatty acid of strain shin9-1T was C18 : 1ω7c. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, diphosphatidylglycerol, phosphatidylmonomethylethanolamine and several uncharacterized lipids. The DNA G+C content was 58.3 mol%. The DNA–DNA relatedness of strain shin9-1T with respect to recognized species of the genus Rhizobium was less than 70 %. Phenotypic characteristics of the novel strain also differed from those of the most closely related species of the genus Rhizobium. On the basis of the phylogenetic inference and phenotypic data, strain shin9-1T should be classified as a representative of a novel species, for which the name Rhizobium ipomoeae sp. nov. is proposed. The type strain is shin9-1T ( = LMG 27163T = KCTC 32148T).

A repeat multi-locus sequence analysis (MLSA) of concatenated nusA, eno, rpoB, gltB, lepA, nuoL and nrdA sequences of strains classified as Achromobacter marplatensis was performed. The results revealed that earlier reported sequence data of the proposed type strain were erroneous, and that the corrected concatenated sequence divergence between the A. marplatensis LMG 26219T ( = CCUG 56371T) sequence type and that of strains of Achromobacter spiritinus was well below the 2.1 % threshold value that delineates species of the genus Achromobacter. These results therefore demonstrated that strains which were classified as A. spiritinus should be reclassified as A. marplatensis and that the name Achromobacter spiritinus should no longer be used. An emendation of the description of Achromobacter marplatensis is warranted.