scholarly journals A Novel Moderately Thermophilic Facultative Methylotroph within the Class Alphaproteobacteria

2021 ◽  
Vol 9 (3) ◽  
pp. 477
Author(s):  
Tajul Islam ◽  
Marcela Hernández ◽  
Amare Gessesse ◽  
J. Colin Murrell ◽  
Lise Øvreås

Methylotrophic bacteria (non-methanotrophic methanol oxidizers) consuming reduced carbon compounds containing no carbon–carbon bonds as their sole carbon and energy source have been found in a great variety of environments. Here, we report a unique moderately thermophilic methanol-oxidising bacterium (strain LS7-MT) that grows optimally at 55 °C (with a growth range spanning 30 to 60 °C). The pure isolate was recovered from a methane-utilizing mixed culture enrichment from an alkaline thermal spring in the Ethiopia Rift Valley, and utilized methanol, methylamine, glucose and a variety of multi-carbon compounds. Phylogenetic analysis of the 16S rRNA gene sequences demonstrated that strain LS7-MT represented a new facultatively methylotrophic bacterium within the order Hyphomicrobiales of the class Alphaproteobacteria. This new strain showed 94 to 96% 16S rRNA gene identity to the two methylotroph genera, Methyloceanibacter and Methyloligella. Analysis of the draft genome of strain LS7-MT revealed genes for methanol dehydrogenase, essential for methanol oxidation. Functional and comparative genomics of this new isolate revealed genomic and physiological divergence from extant methylotrophs. Strain LS7-MT contained a complete mxaF gene cluster and xoxF1 encoding the lanthanide-dependent methanol dehydrogenase (XoxF). This is the first report of methanol oxidation at 55 °C by a moderately thermophilic bacterium within the class Alphaproteobacteria. These findings expand our knowledge of methylotrophy by the phylum Proteobacteria in thermal ecosystems and their contribution to global carbon and nitrogen cycles.

2020 ◽  
Vol 70 (8) ◽  
pp. 4646-4652 ◽  
Author(s):  
Nadezhda V. Agafonova ◽  
Elena N. Kaparullina ◽  
Denis S. Grouzdev ◽  
Nina V. Doronina

Novel aerobic, restricted facultatively methylotrophic bacteria were isolated from buds of English oak (Quercus robur L.; strain DubT) and northern red oak (Quercus rubra L.; strain KrD). The isolates were Gram-negative, asporogenous, motile short rods that multiplied by binary fisson. They utilized methanol, methylamine and a few polycarbon compounds as carbon and energy sources. Optimal growth occurred at 25 °C and pH 7.5. The dominant phospholipids were phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol and phoshatidylglycerol. The major cellular fatty acids of cells were C18 : 1 ω7c, 11-methyl C18 : 1 ω7c and C16 : 0. The major ubiquinone was Q-10. Analysis of 16S rRNA gene sequences showed that the strains were closely related to the members of the genus Hansschlegelia : Hansschlegelia zhihuaiae S113T(97.5–98.0 %), Hansschlegelia plantiphila S1T (97.4–97.6 %) and Hansschlegelia beijingensis PG04T(97.0–97.2 %). The 16S rRNA gene sequence similarity between strains DubT and KrD was 99.7 %, and the DNA–DNA hybridization (DDH) result between the strains was 85 %. The ANI and the DDH values between strain DubT and H. zhihuaiae S113T were 80.1 and 21.5  %, respectively. Genome sequencing of the strain DubT revealed a genome size of 3.57 Mbp and a G+C content of 67.0 mol%. Based on the results of the phenotypic, chemotaxonomic and genotypic analyses, it is proposed that the isolates be assigned to the genus Hansschlegelia as Hansschlegelia quercus sp. nov. with the type strain DubT (=VKM B-3284T=CCUG 73648T=JCM 33463T).


2020 ◽  
Vol 70 (3) ◽  
pp. 2089-2095 ◽  
Author(s):  
Li juan Jia ◽  
Kai shuai Zhang ◽  
Kai Tang ◽  
Jian yu Meng ◽  
Chao Zheng ◽  
...  

A pink-pigmented, Gram-negative, rod-shaped, obligate aerobic bacterial strain, MIMD6T, was isolated from biological soil crusts in PR China. Cells grew at 20–37 °C (optimum, 30 °C), at pH 6–8 (optimum, pH 7) and with 0–1 % (w/v) NaCl (optimum, 0 %). Strain MIMD6T could use methanol or formate as a sole carbon source to grow, and carried methanol dehydrogenase genes mxaF and xoxF, supporting its methylotrophic metabolism. The respiratory quinone was ubiquinone Q-10, the major fatty acids were C18 : 1ω7c (87.3 %), and the major polar lipids were diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, one unknown aminolipid and one unidentified glycolipid. The results of phylogenetic analyses based on the sequences of the 16S rRNA gene, seven housekeeping genes (dnaK, recA, rimO, rpIK, rpmG, rpsR and rpoB) and methanol dehydrogenase genes indicated that strain MIMD6T formed a phylogenetic linage with members of the genus Methylobacterium . Strain MIMD6T was most closely related to Methylobacterium isbiliense DSM 17168T and Methylobacterium nodulans LMG 21967T with 16S rRNA gene sequence similarities of 95.7 and 95.2 %, respectively. The genomic DNA G+C content calculated via draft genome sequencing was 73.0 mol%. The average nucleotide identity and digital DNA–DNA hybridization values between strain MIMD6T and the type strains of other Methylobacterium species were 70.7–82.0 and 24.6–30.0 %, respectively. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain MIMD6T represents a novel species of the genus Methylobacterium , for which the name Methylobacterium crusticola sp. nov. is proposed. The type strain is MIMD6T (=KCTC 52305T=MCCC 1K01311T).


Author(s):  
Kiran Kirdat ◽  
Bhavesh Tiwarekar ◽  
Vipool Thorat ◽  
Shivaji Sathe ◽  
Yogesh Shouche ◽  
...  

Sugarcane Grassy Shoot (SCGS) disease is known to be related to Rice Yellow Dwarf (RYD) phytoplasmas (16SrXI-B group) which are found predominantly in sugarcane growing areas of the Indian subcontinent and South-East Asia. The 16S rRNA gene sequences of SCGS phytoplasma strains belonging to the 16SrXI-B group share 98.07 % similarity with ‘Ca. Phytoplasma cynodontis’ strain BGWL-C1 followed by 97.65 % similarity with ‘Ca. P. oryzae’ strain RYD-J. Being placed distinctly away from both the phylogenetically related species, the taxonomic identity of SCGS phytoplasma is unclear and confusing. We attempted to resolve the phylogenetic positions of SCGS phytoplasma based on the phylogenetic analysis of 16S rRNA gene (>1500 bp), nine housekeeping genes (>3500 aa), core genome phylogeny (>10 000 aa) and OGRI values. The draft genome sequences of SCGS phytoplasma (strain SCGS) and Bermuda Grass White leaf (BGWL) phytoplasma (strain LW01), closely related to ‘Ca. P. cynodontis’, were obtained. The SCGS genome was comprised of 29 scaffolds corresponding to 505 173 bp while LW01 assembly contained 21 scaffolds corresponding to 483 935 bp with the fold coverages over 330× and completeness over 90 % for both the genomes. The G+C content of SCGS was 19.86 % while that of LW01 was 20.46 %. The orthoANI values for the strain SCGS against strains LW01 was 79.42 %, and dDDH values were 22. Overall analysis reveals that SCGS phytoplasma forms a distant clade in RYD group of phytoplasmas. Based on phylogenetic analyses and OGRI values obtained from the genome sequences, a novel taxon ‘Candidatus Phytoplasma sacchari’ is proposed.


Author(s):  
Xue-Gong Li ◽  
Jin Lin ◽  
Shi-Jie Bai ◽  
Jie Dai ◽  
Ze-Xi Jiao ◽  
...  

A novel moderately thermophilic, anaerobic, heterotrophic bacterium (strain SY095T) was isolated from a hydrothermal vent chimney located on the Southwest Indian Ridge at a depth of 2730 m. Cells were Gram-stain-positive, motile, straight to slightly curved rods forming terminal endospores. SY095T was grown at 45–60 °C (optimum 50–55 °C), pH 6.0–7.5 (optimum 7.0), and in a salinity of 1–4.5 % (w/v) NaCl (optimum 2.5 %). Substrates utilized by SY095T included fructose, glucose, maltose, N-acetyl glucosamine and tryptone. Casamino acid and amino acids (glutamate, glutamine, lysine, methionine, serine and histidine) were also utilized. The main end products from glucose fermentation were acetate, H2 and CO2. Elemental sulphur, sulphate, thiosulphate, sulphite, fumarate, nitrate, nitrite and Fe(III) were not used as terminal electron acceptors. The predominant cellular fatty acids were C14 : 0 (60.5%) and C16 : 0 (7.6 %). The main polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, five unidentified phospholipids and two unidentified aminophospholipids. No respiratory quinones were detected. The chromosomal DNA G+C content was 30.8 mol%. The results of phylogenetic analysis of the 16S rRNA gene sequences indicated that SY095T was closely related to Crassaminicella profunda Ra1766HT (95.8 % 16S rRNA gene sequence identity). SY095T exhibited 78.1 % average nucleotide identity (ANI) to C. profunda Ra1766HT. The in silico DNA–DNA hybridization (DDH) value indicated that SY095T shared 22.7 % DNA relatedness with C. profunda Ra1766HT. On the basis of its phenotypic, genotypic and phylogenetic characteristics, SY095T is suggested to represent a novel species of the genus Crassaminicella , for which the name Crassaminicella thermophila sp. nov. is proposed. The type strain is SY095T (=JCM 34213=MCCC 1K04191). An emended description of the genus Crassaminicella is also proposed.


2014 ◽  
Vol 64 (Pt_3) ◽  
pp. 989-999 ◽  
Author(s):  
Hisako Hirayama ◽  
Mariko Abe ◽  
Masayuki Miyazaki ◽  
Takuro Nunoura ◽  
Yasuo Furushima ◽  
...  

A novel methane-oxidizing bacterium, strain IT-9T, was isolated from a shallow submarine hydrothermal system occurring in a coral reef in Japan. Strain IT-9T was a Gram-negative, aerobic, motile, coccoid or oval-shaped bacterium with the distinctive intracytoplasmic membrane arrangement of a type I methanotroph. Strain IT-9T was a moderately thermophilic, obligate methanotroph that grew on methane and methanol at 30–55 °C (optimum 45–50 °C). The strain possessed the particulate methane monooxygenase (pMMO). The ribulose monophosphate pathway was operative for carbon assimilation. NaCl was required for growth within a concentration range of 1–5 % (optimum 3 %). The hao gene encoding hydroxylamine oxidoreductase (HAO) involved in nitrification was detected by a PCR experiment. The major phospholipid fatty acids were C16 : 0 and C18 : 1ω7c. The major isoprenoid quinone was Q-8. The DNA G+C content was 66.0 mol%. The 16S rRNA gene sequence of strain IT-9T was only moderately related to the sequences of members of the closest genera Methylohalobius (94.1 % similarity) and Methylothermus (91.7–91.9 % similarity); however, those sequences formed a deeply branching monophyletic group within the order Methylococcales . Phylogenies based on 16S rRNA gene sequences, deduced partial PmoA sequences and deduced partial Hao sequences and physiological and chemotaxonomic characteristics revealed that strain IT-9T represents a novel species of a new genus, for which the name Methylomarinovum caldicuralii gen. nov., sp. nov. is proposed. The type strain of Methylomarinovum caldicuralii is IT-9T ( = JCM 13666T = DSM 19749T). In addition, we propose a new family, Methylothermaceae fam. nov., in the order Methylococcales , to accommodate the genera Methylothermus , Methylohalobius and Methylomarinovum. The genera Methylothermus and Methylohalobius have been recognized as being distinct from other genera in the methane-oxidizing order Methylococcales in the class Gammaproteobacteria . These genera form a distinctive monophyletic lineage within the order on the basis of 16S rRNA gene sequence phylogeny. This seems consistent with their distinctive physiological traits; the genus Methylothermus includes the most thermophilic species, and the genus Methylohalobius includes the most halophilic species, within the order. Although these two genera include only three species at the time of writing, similar sequences of 16S rRNA genes and pmoA genes encoding pMMO have been detected in a geothermal area or deep-sea hydrothermal vent fields by studies using culture-independent techniques. This suggests that unknown methanotrophs of this lineage inhabit various extreme environments.


2012 ◽  
Vol 62 (Pt_7) ◽  
pp. 1647-1652 ◽  
Author(s):  
Akio Tani ◽  
Nurettin Sahin ◽  
Kazuhide Kimbara

A pink-pigmented, facultatively methylotrophic bacterium, strain 35aT, was isolated from the leaves of Oxalis corniculata. Cells of strain 35aT were Gram-reaction-negative, motile, non-spore-forming rods. The highest 16S rRNA gene pairwise sequence similarities for strain 35aT were found with the strains of Methylobacterium iners 5317S-33T (96.7 %), ‘Methylobacterium soli’ YIM 48816 (96.6 %) and Methylobacterium jeotgali S2R03-9T (96.3 %). 16S rRNA gene sequence similarities with the type strains of all other recognized species of the genus Methylobacterium were below 96 %. Major cellular fatty acids were C18 : 1ω7c, C18 : 0 and C16 : 0. The results of DNA–DNA hybridization experiments, analysis of cpn60 gene sequences, fatty acid profiles, whole-cell MALDI-TOF/MS spectral pattern analysis, and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain 35aT from its nearest phylogenetic neighbours. Strain 35aT is therefore considered to represent a novel species within the genus Methylobacterium , for which the name Methylobacterium oxalidis sp. nov. is proposed. The type strain is 35aT ( = DSM 24028T = NBRC 107715T).


2011 ◽  
Vol 61 (11) ◽  
pp. 2646-2653 ◽  
Author(s):  
Hisako Hirayama ◽  
Yohey Suzuki ◽  
Mariko Abe ◽  
Masayuki Miyazaki ◽  
Hiroko Makita ◽  
...  

A novel methane-oxidizing bacterium, strain HTM55T, was isolated from subsurface hot aquifer water from a Japanese gold mine. Strain HTM55T was a Gram-negative, aerobic, motile, coccoid bacterium with a single polar flagellum and the distinctive intracytoplasmic membrane arrangement of a type I methanotroph. Strain HTM55T was a moderately thermophilic, obligate methanotroph that grew on methane and methanol at 37–65 °C (optimum 55–60 °C). The isolate grew at pH 5.2–7.5 (optimum 5.8–6.3) and with 0–1 % NaCl (optimum 0–0.3 %). The ribulose monophosphate pathway was operative for carbon assimilation. The DNA G+C content was 54.4 mol% and the major fatty acids were C16 : 0 (52.0 %) and C18 : 1ω7c (34.8 %). Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain HTM55T was closely related to Methylothermus thermalis MYHTT (99.2 % 16S rRNA gene sequence similarity), which is within the class Gammaproteobacteria. However, DNA–DNA relatedness between strain HTM55T and Methylothermus thermalis MYHTT was ≤39 %. On the basis of distinct phylogenetic, chemotaxonomic and physiological characteristics, strain HTM55T represents a novel species of the genus Methylothermus, for which the name Methylothermus subterraneus sp. nov. is proposed. The type strain is HTM55T ( = JCM 13664T = DSM 19750T).


2007 ◽  
Vol 57 (7) ◽  
pp. 1473-1477 ◽  
Author(s):  
Keiko Watanabe ◽  
Norio Nagao ◽  
Shuich Yamamoto ◽  
Tatsuki Toda ◽  
Norio Kurosawa

A Gram-negative, rod-shaped, spore-forming and moderately thermophilic bacterium, strain KWC4T, was isolated from a composting reactor. Cells of strain KWC4T were 2.0–5.0 μm long and 0.5–0.7 μm in diameter. Strain KWC4T grew aerobically at 32–61 °C, with optimal growth occurring at 50 °C. It grew at pH 5.6–10.1, with optimal growth at around pH 9.0. The optimum NaCl concentration for growth was almost 0 % (w/v), but strain KWC4T was moderately halotolerant and was able to grow at NaCl concentrations up to 4.4 % (w/v). The DNA G+C content of strain KWC4T was 60.0 mol%. The major fatty acids were iso-16 : 0 (39.0 %) and anteiso-15 : 0 (33.3 %). Based on 16S rRNA gene sequence similarity data, strain KWC4T belonged to the genus Thermobacillus and was related to Thermobacillus xylanilyticus. However, strain KWC4T had a 38 bp insertion sequence located near the 3′ end of its 16S rRNA gene that was not present in T. xylanilyticus. The 16S rRNA gene sequence similarity value between strain KWC4T and T. xylanilyticus was 95.7 %. The DNA–DNA hybridization value between strain KWC4T and T. xylanilyticus strain XET was 66 %. On the basis of phenotypic and genotypic evidence, strain KWC4T (=DSM 18247T=JCM 13945T) is the type strain of a novel species, for which the name Thermobacillus composti sp. nov. is proposed.


2013 ◽  
Vol 63 (Pt_9) ◽  
pp. 3287-3292 ◽  
Author(s):  
Akio Tani ◽  
Nurettin Sahin

Pink-pigmented, facultatively methylotrophic bacteria, strains 87eT and 99bT, were isolated from the bryophytes Haplocladium microphyllum and Brachythecium plumosum, respectively. The cells of both strains were Gram-reaction-negative, motile, non-spore-forming rods. On the basis of 16S rRNA gene sequence similarity, strains 87eT and 99bT were found to be related to Methylobacterium organophilum ATCC 27886T (97.1 % and 97.7 %, respectively). Strains 87eT and 99bT showed highest 16S rRNA gene similarity to Methylobacterium gnaphalii 23eT (98.3 and 99.0 %, respectively). The phylogenetic similarities to all other species of the genus Methylobacterium with validly published names were less than 97 %. Major cellular fatty acids of both strains were C18 : 1ω7c and C18 : 0. The results of DNA–DNA hybridization, phylogenetic analyses based on 16S rRNA and cpn60 gene sequences, fatty acid profiles, whole-cell matrix-assisted, laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis, and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strains 87eT and 99bT from their phylogenetically closest relatives. We propose that strains 87eT and 99bT represent novel species within the genus Methylobacterium , for which the names Methylobacterium haplocladii sp. nov. (type strain 87eT = DSM 24195T = NBRC 107714T) and Methylobacterium brachythecii sp. nov. (type strain 99bT = DSM 24105T = NBRC 107710T) are proposed.


2015 ◽  
Vol 65 (Pt_8) ◽  
pp. 2410-2419 ◽  
Author(s):  
Yonghui Zeng ◽  
Vadim Selyanin ◽  
Martin Lukeš ◽  
Jason Dean ◽  
David Kaftan ◽  
...  

A red-pigmented, bacteriochlorophyll (BChl) a-producing strain, AP64T, was isolated previously from the freshwater Swan Lake located in the western Gobi Desert. Based on its 16S rRNA gene sequence identity (96.1 %) to the type strain Gemmatimonas aurantiaca T-27T, the new isolate was tentatively classified as a member of the bacterial phylum Gemmatimonadetes. Here, we report its formal description and polyphasic characterization. Strain AP64T grew best on agar media under 9.8–15.2 % atmospheric oxygen. The cells were rods, dividing by symmetrical or asymmetrical binary fission. Budding structures were also observed. Its genomic DNA G+C content was 64.4 % (from the draft genome sequence). Phylogenetic analysis based on the 16S rRNA gene sequence clearly separated AP64T from related species. Its genotypic differentiation from phylogenetically close relatives was further supported by performing in silico DNA–DNA hybridization and calculating average nucleotide identity, whereas the high percentage (67.3 %) of shared conserved proteins between strain AP64T and Gemmatimonas aurantiaca T-27T supports the classification of the two strains into the same genus. Strain AP64T contained C16 : 1, C14 : 1 and C18 : 1ω9c as predominant fatty acids. The main respiratory quinone was menaquinone 8 (MK-8). The most distinctive feature of strain AP64T was the presence of fully functional purple bacterial photosynthetic reaction centres. The main CO2-fixation pathways were absent. Strain AP64T was capable of growth and BChl production in constant darkness. Thus, strain AP64T is a facultatively photoheterotrophic organism. It represents a novel species of the genus Gemmatimonas, for which the name Gemmatimonas phototrophica sp. nov. is proposed. The type strain is AP64T ( = DSM 29774T = MCCC 1K00454T). Emended descriptions of the genus Gemmatimonas and Gemmatimonas aurantiaca are also provided.


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