scholarly journals Identification of a Toluene-Degrading Bacterium from a Soil Sample through H218O DNA Stable Isotope Probing

2011 ◽  
Vol 77 (17) ◽  
pp. 5995-5999 ◽  
Author(s):  
Angela Woods ◽  
Maribeth Watwood ◽  
Egbert Schwartz

ABSTRACTDNA stable isotope probing (DNA-SIP) with H218O was used to identify a toluene-degrading bacterium in soil amended with 48 ppm toluene. After quantification of toluene degradation rates in soil, DNA was extracted from soil incubated with H218O, H216O, H216O and 48 ppm toluene, or H218O and 48 ppm toluene. A single DNA band formed along a cesium chloride gradient after isopycnic centrifugation of extracts from soils incubated with H216O. With extracts from soils to which only H218O was added, two distinct DNA bands formed, while three bands formed when DNA extracted from soil incubated with both H218O and toluene was analyzed. We suggest that this third band formed because toluene does not contain any oxygen atoms and toluene-degrading organisms had to transfer oxygen atoms from H218O into metabolic intermediates to form nucleic acidsde novo. We extracted the third DNA band and amplified a large fraction of the bacterial 16S rRNA gene. Direct sequencing of the PCR product obtained from the labeled DNA, as well as cloned 16S rRNA amplicons, identified a known toluene degrader,Rhodococcus jostiiRHA1. A toluene-degrading bacterial strain was subsequently isolated from soil and shown to beRhodococcus jostiiRHA1. Finally, quantitative real-time PCR analysis showed that the abundance of the 16S rRNA gene ofRhodococcus jostiiRHA1 increased in soil after toluene exposure but not in soils from which toluene was withheld. This study indicates that H218O DNA-SIP can be a useful method for identifying pollutant-degrading bacteria in soil.

2015 ◽  
Vol 65 (Pt_5) ◽  
pp. 1550-1555 ◽  
Author(s):  
Hyun Mi Jin ◽  
Hye Im Jeong ◽  
Che Ok Jeon

A Gram-stain-negative heterotrophic bacterium, designated GSD6T, capable of growth on aliphatic hydrocarbons as a sole carbon and energy source, was isolated from sea-tidal flat sediment of the Yellow Sea, South Korea. Cells were facultatively aerobic, catalase- and oxidase-positive, motile rods with a single polar flagellum. Growth of strain GSD6T was observed at 4–37 °C (optimum 30 °C), at pH 5.5–9.0 (optimum pH 6.5–7.5) and in the presence of 1–9 % (w/v) NaCl (optimum 2 %). Strain GSD6T contained ubiquinone-8 (Q-8) as the sole isoprenoid quinone and summed feature 3 (comprising C16 : 1ω7c and/or iso-C15 : 0 2-OH), C16 : 0, C18 : 1ω7c, C17 : 0 10-methyl and C17 : 1ω8c as the major fatty acids. Phosphatidylethanolamine and phosphatidylglycerol were identified as the major polar lipids. The G+C content of the genomic DNA was 44.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GSD6T formed a phylogenetic lineage with members of the genus Aliiglaciecola . Strain GSD6T was most closely related to Aliiglaciecola lipolytica E3T with a 16S rRNA gene sequence similarity of 97.4 %, but their DNA–DNA hybridization value was 39.1±7.1 %. On the basis of phenotypic, chemotaxonomic and molecular features, strain GSD6T represents a novel species of the genus Aliiglaciecola , for which the name Aliiglaciecola aliphaticivorans sp. nov. is proposed. The type strain is GSD6T ( = KACC 18129T = JCM 30133T). An emended description of the genus Aliiglaciecola is also proposed.


2014 ◽  
Vol 64 (Pt_11) ◽  
pp. 3768-3774 ◽  
Author(s):  
Naysim LO ◽  
Hyo Jung Kang ◽  
Che Ok Jeon

A Gram-staining-negative, facultatively aerobic bacterium, designated SM-2T, was isolated from a sea-tidal flat of Yellow Sea, South Korea. Cells were catalase- and oxidase-positive motile rods with a single polar flagellum. Growth of strain SM-2T was observed at 10–37 °C (optimum, 25–30 °C), at pH 5.5–8.5 (optimum, pH 7.0–7.5) and in the presence of 0–11 % (w/v) NaCl (optimum, 2 %). Strain SM-2T contained ubiquinone-8 (Q-8) as the sole isoprenoid quinone and C17 : 1ω8c, summed feature 3 (comprising C16 : 1ω7c and/or iso-C15 : 0 2-OH), C17 : 0 and C18 : 1ω7c as the major fatty acids. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified lipid were identified as the major cellular polar lipids. The G+C content of the genomic DNA was 52.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SM-2T formed a tight phyletic lineage with Zhongshania antarctica ZS5-23T, Zhongshania guokunii ZS6-22T and Spongiibacter borealis CL-AS9T, but that S. borealis CL-AS9T was distinct from other species of the genus Spongiibacter . Based on 16S rRNA gene sequence similarities, strain SM-2T was most closely related to S. borealis CL-AS9T, Z. antarctica ZS5-23T and Z. guokunii ZS6-22T, with similarities of 99.5 %, 98.9 % and 98.7 %, respectively, but the DNA–DNA hybridization values among these species were clearly lower than 70 %. On the basis of chemotaxonomic data and molecular properties, we propose strain SM-2T represents a novel species of the genus Zhongshania with the name Zhongshania aliphaticivorans sp. nov. (type strain SM-2T = KACC 18120T = JCM 30138T). We also propose the transfer of Spongiibacter borealis Jang et al. 2011 to the genus Zhongshania as Zhongshania borealis comb. nov. (type strain CL-AS9T = KCCM 90094T = JCM 17304T).


2014 ◽  
Vol 64 (Pt_6) ◽  
pp. 2017-2022 ◽  
Author(s):  
Tao Gu ◽  
Li Na Sun ◽  
Jun Zhang ◽  
Xin Hua Sui ◽  
Shun Peng Li

A Gram-stain-negative, non-motile, pale yellow, rod-shaped bacterial strain, YW14T, was isolated from soil and its taxonomic position was investigated by a polyphasic study. Strain YW14T did not form nodules on three different legumes, and the nodD and nifH genes were not detected by PCR. Strain YW14T contained Q-10 as the predominant ubiquinone. The major cellular fatty acid was C18 : 1ω7c. Phylogenetic analyses based on 16S rRNA gene sequences and seven housekeeping gene sequences (recA, atpD, glnII, gyrB, rpoB, dnaK and thrC) showed that strain YW14T belonged to the genus Rhizobium . Strain YW14T showed 16S rRNA gene sequence similarity of 93.4–97.3 % to the type strains of recognized species of the genus Rhizobium . DNA–DNA relatedness between strain YW14T and the type strains of Rhizobium sullae IS123T and Rhizobium yanglingense CCBAU 71623T was 19.6–25.7 %, indicating that strain YW14T was distinct from them genetically. Strain YW14T could also be differentiated from these phylogenetically related species of the genus Rhizobium by various phenotypic properties. On the basis of phenotypic properties, phylogenetic distinctiveness and genetic data, strain YW14T is considered to represent a novel species of the genus Rhizobium , for which the name Rhizobium flavum sp. nov. is proposed. The type strain is YW14T ( = KACC 17222T = CCTCC AB2013042T).


2015 ◽  
Vol 81 (14) ◽  
pp. 4607-4615 ◽  
Author(s):  
Xiaoqing Wang ◽  
Christine E. Sharp ◽  
Gareth M. Jones ◽  
Stephen E. Grasby ◽  
Allyson L. Brady ◽  
...  

ABSTRACTThe exopolysaccharides (EPSs) produced by some bacteria are potential growth substrates for other bacteria in soil. We used stable-isotope probing (SIP) to identify aerobic soil bacteria that assimilated the cellulose produced byGluconacetobacter xylinusor the EPS produced byBeijerinckia indica. The latter is a heteropolysaccharide comprised primarily ofl-guluronic acid,d-glucose, andd-glycero-d-mannoheptose.13C-labeled EPS and13C-labeled cellulose were purified from bacterial cultures grown on [13C]glucose. Two soils were incubated with these substrates, and bacteria actively assimilating them were identified via pyrosequencing of 16S rRNA genes recovered from13C-labeled DNA. Cellulose C was assimilated primarily by soil bacteria closely related (93 to 100% 16S rRNA gene sequence identities) to known cellulose-degrading bacteria. However,B. indicaEPS was assimilated primarily by bacteria with low identities (80 to 95%) to known species, particularly by different members of the phylumPlanctomycetes. In one incubation, members of thePlanctomycetesmade up >60% of all reads in the labeled DNA and were only distantly related (<85% identity) to any described species. Although it is impossible with SIP to completely distinguish primary polysaccharide hydrolyzers from bacteria growing on produced oligo- or monosaccharides, the predominance ofPlanctomycetessuggested that they were primary degraders of EPS. Other bacteria assimilatingB. indicaEPS included members of theVerrucomicrobia, candidate division OD1, and theArmatimonadetes. The results indicate that some uncultured bacteria in soils may be adapted to using complex heteropolysaccharides for growth and suggest that the use of these substrates may provide a means for culturing new species.


2011 ◽  
Vol 78 (3) ◽  
pp. 628-637 ◽  
Author(s):  
Tony Gutierrez ◽  
Peter D. Nichols ◽  
William B. Whitman ◽  
Michael D. Aitken

ABSTRACTA marine bacterium, designated strain MCTG13d, was isolated from a laboratory culture of the dinoflagellateLingulodinium polyedrumCCAP1121/2 by enrichment with polycyclic aromatic hydrocarbons (PAHs) as the sole carbon source. Based on 16S rRNA gene sequence comparisons, the strain was most closely related toPorticoccus litoralisIMCC2115T(96.5%) and to members of the generaMicrobulbifer(91.4 to 93.7%) andMarinimicrobium(90.4 to 92.0%). Phylogenetic trees showed that the strain clustered in a distinct phyletic line in the classGammaproteobacteriafor whichP. litoralisis presently the sole cultured representative. The strain was strictly aerobic, rod shaped, Gram negative, and halophilic. Notably, it was able to utilize hydrocarbons as sole sources of carbon and energy, whereas sugars did not serve as growth substrates. The predominant isoprenoid quinone of strain MCTG13d was Q-8, and the dominant fatty acids were C16:1ω7c, C18:1ω7c, and C16:0. DNA G+C content for the isolate was 54.9 ± 0.42 mol%. Quantitative PCR primers targeting the 16S rRNA gene of this strain showed that this organism was common in other laboratory cultures of marine phytoplankton. On the basis of phenotypic and genotypic characteristics, strain MCTG13d represents a novel species ofPorticoccus, for which the namePorticoccus hydrocarbonoclasticussp. nov. is proposed. The discovery of this highly specialized hydrocarbon-degrading bacterium living in association with marine phytoplankton suggests that phytoplankton represent a previously unrecognized biotope of novel bacterial taxa that degrade hydrocarbons in the ocean.


2015 ◽  
Vol 65 (Pt_1) ◽  
pp. 274-279 ◽  
Author(s):  
Milán Farkas ◽  
András Táncsics ◽  
Balázs Kriszt ◽  
Tibor Benedek ◽  
Erika M. Tóth ◽  
...  

A floc-forming, Gram-stain-negative, petroleum hydrocarbon-degrading bacterial strain, designated BucT, was isolated from a petroleum hydrocarbon-contaminated site in Hungary. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BucT formed a distinct phyletic lineage within the genus Zoogloea . Its closest relative was found to be Zoogloea caeni EMB43T (97.2 % 16S rRNA gene sequence similarity) followed by Zoogloea oryzae A-7T (95.9 %), Zoogloea ramigera ATCC 19544T (95.5 %) and Zoogloea resiniphila DhA-35T (95.4 %). The level of DNA–DNA relatedness between strain BucT and Z. caeni EMB43T was 31.6 %. Cells of strain BucT are facultatively aerobic, rod-shaped, and motile by means of a polar flagellum. The strain grew at temperatures of 5–35 °C (optimum 25–28 °C), and at pH 6.0–9.0 (optimum 6.5–7.5). The predominant fatty acids were C16 : 0, C10 : 0 3-OH, C12 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The major respiratory quinone was ubiquinone-8 (Q-8) and the predominant polar lipid was phosphatidylethanolamine. The genomic DNA G+C content was 63.2 mol%. On the basis of the chemotaxonomic, molecular and phenotypic data, isolate BucT is considered to represent a novel species of the genus Zoogloea , for which the name Zoogloea oleivorans sp. nov. is proposed. The type strain is BucT ( = DSM 28387T = NCAIM B 02570T).


2015 ◽  
Vol 65 (Pt_2) ◽  
pp. 570-577 ◽  
Author(s):  
Masao Sakai ◽  
Daigo Deguchi ◽  
Akifumi Hosoda ◽  
Tomohiro Kawauchi ◽  
Makoto Ikenaga

A thermophilic, agar-degrading bacterium, strain FAB2T, was isolated from sewage sludge compost. According to phylogenetic analysis based on 16S rRNA gene sequences, strain FAB2T belonged to the family Paenibacillaceae within the phylum Firmicutes . However, FAB2T was different enough at the genus level from closely related species. The percentages of 16S rRNA gene sequence similarity with related organisms were 90.4 % for Thermobacillus xylanilyticus , 91.8 % for Paenibacillus barengoltzii , 89.4 % for Cohnella lupini , 90.1 % for Fontibacillus aquaticus , and 89.0 % for Saccharibacillus sacchari . Morphological and physiological analyses revealed that the strain was motile, rod-shaped, Gram-stain-positive, aerobic and able to form oval endospores in swollen sporangia. Ammonium was required as a nitrogen source while nitrate, nitrite, urea and glutamate were not utilized. Catalase and oxidase activities were weakly positive and positive, respectively. The bacterium grew in the temperature range of 50–65 °C and in media with pH 7.5 to 9.0. Optimal growth occurred at 60 °C and pH 8.0–8.6. Growth was inhibited at pH≤7.0 and NaCl concentrations ≥2.5 % (w/v). In chemotaxonomic characterization, MK-7 was identified as the dominant menaquinone. Major fatty acids were iso-C16 : 0 and C16 : 0. Dominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Phosphatidylcholine was present in a moderate amount. The diamino acid in the cell wall was meso-diaminopimelic acid. The G+C content of the genomic DNA was 49.5 mol% in a nucleic acid study. On the basis of genetic and phenotypic characteristics, strain FAB2T ( = NBRC 109510T = KCTC 33130T) showed characteristics suitable for classification as the type strain of a novel species of a new genus in the family Paenibacillaceae , for which the name Ammoniibacillus agariperforans gen. nov., sp. nov. is proposed.


2014 ◽  
Vol 64 (Pt_6) ◽  
pp. 2047-2052 ◽  
Author(s):  
Li-Na Sun ◽  
Jun Zhang ◽  
Fen-Fen Gong ◽  
Xiang Wang ◽  
Gang Hu ◽  
...  

The taxonomic status of a carbendazim-degrading strain, mbc-2T, isolated from soil under the long-term application of carbendazim in China was determined by means of a polyphasic study. The cells were Gram-stain-positive, motile and rod-shaped. Strain mbc-2T grew optimally at pH 7.0, 30–35 °C and in the presence of 1 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain mbc-2T fell within the genus Nocardioides , forming a coherent cluster with the type strain of Nocardioides hankookensis , with which it exhibited 16S rRNA gene sequence similarity values of 97.9 %. The chemotaxonomic properties of strain mbc-2T were consistent with those of the genus Nocardioides : the cell-wall peptidoglycan type was based on ll-2,6-diaminopimelic acid, the predominant menaquinone was MK-8 (H4) and the major fatty acid was iso-C16 : 0. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, unknown phospholipids and an unknown aminolipid. The DNA G+C content was 72 mol%. Strain mbc-2T exhibited DNA–DNA relatedness values of 12.5±1.5 %, 23.7±2.7 % and 26.3±3.2 % with respect to Nocardioides hankookensis DS-30T, Nocardioides aquiterrae GW-9T and Nocardioides pyridinolyticus OS4T. On the basis of the data obtained, strain mbc-2T represents a novel species of the genus Nocardioides , for which the name Nocardioides soli sp. nov. is proposed. The type strain is mbc-2T ( = KACC 17152T = CCTCC AB 2012934T).


2013 ◽  
Vol 63 (Pt_12) ◽  
pp. 4477-4483 ◽  
Author(s):  
Yunhui Zhang ◽  
Kaihao Tang ◽  
Xiaochong Shi ◽  
Xiao-Hua Zhang

A Gram-stain-negative, strictly aerobic, yellow-pigmented, rod-shaped and N-acylhomoserine lactone-degrading bacterium, designated strain Th78T, was isolated from the intestine of a cultured flounder (Paralichthys olivaceus). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain Th78T belonged to the genus Flaviramulus (family Flavobacteriaceae ) and showed the highest 16S rRNA gene sequence similarity to Flaviramulus basaltis H35T (96.70 %). Optimal growth occurred in the presence of 2–3 % (w/v) NaCl, at pH 7.0–8.0 and at 28 °C. The major fatty acids were iso-C15 : 0 3-OH, iso-C15 : 1 G and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine, one unidentified aminolipid and three unidentified polar lipids. Menaquinone 6 (MK-6) was the only respiratory quinone. The DNA G+C content of strain Th78T was 31.5 mol%. On the basis of polyphasic analysis, strain Th78T is considered to represent a novel species of the genus Flaviramulus , for which the name Flaviramulus ichthyoenteri sp. nov. is proposed. The type strain is Th78T ( = JCM 18634T = KCTC 32142T = DSM 26285T). Emended descriptions of the genus Flaviramulus and Flaviramulus basaltis are also proposed.


2013 ◽  
Vol 63 (Pt_8) ◽  
pp. 3075-3078 ◽  
Author(s):  
Shigeto Otsuka ◽  
Taku Suenaga ◽  
Hoan Thi Vu ◽  
Hiroyuki Ueda ◽  
Akira Yokota ◽  
...  

The taxonomic properties of strain DC2c-G4T, a Gram-staining-negative, ovoid, gellan-gum-degrading bacterial isolate, were examined. Phylogenetic analysis based on 16S rRNA gene sequences identified this isolate as a member of the phylum Verrucomicrobia and closest to the genus Prosthecobacter . The 16S rRNA gene sequence similarities between this isolate and any of the type strains of species of the genus Prosthecobacter were less than 95 %. In addition, the absence of a single prostheca and the predominant menaquinone MK-7(H2) supported the differentiation of this isolate from the genus Prosthecobacter . Here, we propose Brevifollis gellanilyticus gen. nov., sp. nov. to accommodate the isolate. The type strain of the type species is DC2c-G4T ( = NBRC 108608T = CIP 110457T).


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