Horizontal Gene Transfer of Genes Encoding Copper-Containing Membrane-Bound Monooxygenase (CuMMO) and Soluble Di-iron Monooxygenase (SDIMO) in Ethane- and Propane-Oxidizing Rhodococcus Bacteria.

The families of copper-containing membrane-bound monooxygenases (CuMMOs) and soluble di-iron monooxygenases (SDIMOs) are involved not only in methane oxidation but also in short-chain alkane oxidation. Here, we describe Rhodococcus sp. strain ZPP, a bacterium able to grow with ethane or propane as the sole carbon and energy source, and report on the horizontal gene transfer (HGT) of actinobacterial hydrocarbon monooxygenases (HMOs) of the CuMMO family and the sMMO (soluble methane monooxygenase)-like SDIMO in the genus Rhodococcus. The key function of HMO in strain ZPP for propane oxidation was verified by allylthiourea inhibition. The HMO genes (designated hmoCAB) and those encoding sMMO-like SDIMO (designated smoXYB1C1Z) are located on a linear megaplasmid (pRZP1) of strain ZPP. Comparative genomic analysis of similar plasmids indicated the mobility of these plasmids within the genus Rhodococcus. The plasmid pRZP1 in strain ZPP could be conjugatively transferred to a recipient Rhodococcus erythropolis strain in a mating experiment and showed similar ethane- and propane-consuming activities. Finally, our findings demonstrate that the horizontal transfer of plasmid-based CuMMO and SDIMO genes confers the ability to use ethane and propane on the recipient. IMPORTANCE CuMMOs and SDIMOs initiate the aerobic oxidation of alkanes in bacteria. Here, the supposition that horizontally transferred plasmid-based CuMMO and SDIMO genes confer on the recipient similar abilities to use ethane and propane was proposed and confirmed in Rhodococcus. This study is a living example of HGT of CuMMOs and SDIMOs and outlines the plasmid-borne properties responsible for gaseous alkane degradation. Our results indicate that plasmids can support the rapid evolution of enzyme-mediated biogeochemical processes.

Description of Fimbriimonas ginsengisoli gen. nov., sp. nov. within the Fimbriimonadia class nov., of the phylum Armatimonadetes.

Strain Gsoil 348(T) was isolated from a ginseng field soil sample by selecting micro-colonies from one-fifth strength modified R2A agar medium after a long incubation period. 16S rRNA gene sequence analysis indicated that the strain is related to members of the phylum Armatimonadetes (formerly called candidate phylum OP10). Strain Gsoil 348(T) is mesophilic, strictly aerobic, non-motile and rod-shaped. It only grows in low nutrient media. The major respiratory quinones are menaquinones MK-11 and MK-10, and the main fatty acids are iso-C(15:0), iso-C(17:0), C(16:0) and C(16:1) ω11c. The G+C content is 61.4 mol%. The 16S rRNA gene sequences in public databases belonging to the phylum Armatimonadetes were clustered here into 6 groups. Five of these groups constituted a coherent cluster distinct from the sequences of other phyla in phylogenetic trees that were constructed using multiple-outgroup sequences from 49 different phyla. On the basis of polyphasic taxonomic analyses, it is proposed that strain Gsoil 348(T) (= KACC 14959(T) = JCM 17079(T)) should be placed in Fimbriimonas ginsengisoli gen. nov., sp. nov., as the cultured representative of the Fimbriimonadia class. nov., corresponding with Group 4 of the phylum Armatimonadetes.

Joostella marina gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from the East Sea.

A Gram-negative, non-spore-forming, non-motile, yellow-pigmented, strictly aerobic bacterial strain, designated En5(T), was isolated from the East Sea of Korea and was subjected to a polyphasic taxonomy study. Strain En5(T) grew optimally at 30 degrees C, in the presence of 1-3 % (w/v) NaCl and at pH 5.3-7.6. The major respiratory lipoquinone was MK-6 and the major fatty acids were iso-C(15 : 0), iso-C(17 : 0) 3-OH and iso-C(17 : 1)omega9c. The DNA G+C content of strain En5(T) was 30.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain En5(T) formed a distinct evolutionary lineage within the family Flavobacteriaceae and shared 93 % sequence similarity with the type strains of both Galbibacter mesophilus and Zhouia amylolytica. On the basis of its phenotypic and phylogenetic properties, strain En5(T) is suggested to represent a novel species of a new genus in the family Flavobacteriaceae, for which the name Joostella marina gen. nov., sp. nov. is proposed. The type strain is En5(T) (=KCTC 12518(T)=DSM 19592(T)=CGMCC 1.6973(T)).

Arthrobacter subterraneus sp. nov., isolated from deep subsurface water of the South Coast of Korea.

Strain CH7T, a pale yellow-pigmented bacterium and new isolate from deep subsurface water of the South Coast of Korea, was subjected to a polyphasic taxonomic study. CH7T grew between 5 and 37 degrees C, pH 5.3-10.5, and tolerated up to 13% NaCl. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain CH7T was associated with the genus Arthrobacter and phylogenetically closely related to the type strains Arthrobacter tumbae (99.4%) and Arthrobacter parietis (99.1%). However, DNA-DNA hybridization experiments revealed 2.1% and 12% between strain CH7T and Arthrobacter tumbae and Arthrobacter parietis, respectively. Thus, the phenotypic and phylogenetic differences suggested that CH7T should be placed in the genus Arthrobacter as a novel species, for which the name Arthrobacter subterraneus sp. nov. is proposed. In addition, the type strain for the new species is CH7T (=KCTC 9997T=DSM 17585T).

Pseudoalteromonas marina sp. nov., a marine bacterium isolated from tidal flats of the Yellow Sea, and reclassification of Pseudoalteromonas sagamiensis as Algicola sagamiensis comb. nov.

Two Gram-negative, motile and strictly aerobic marine bacteria were isolated from a tidal flat sediment sample obtained from Dae-Chun, Chung-Nam, Korea. They were preliminarily identified as Pseudoalteromonas-like bacteria, based on 16S rRNA gene sequence analysis showing nearly identical sequences (>99.7 % sequence similarity) and the highest similarity (98.4 %) to the species Pseudoalteromonas undina. Some phenotypic features of the newly isolated strains were similar to those of members of the genus Pseudoalteromonas, but several physiological and chemo-taxonomical properties readily distinguished the new isolates from previously described species. DNA-DNA hybridization with type strains of phylogenetically closely related species demonstrated that the isolates represent a novel Pseudoalteromonas species, for which the name Pseudoalteromonas marina sp. nov. is proposed, with the type strain mano4(T) (=KCTC 12242(T)=DSM 17587(T)). In addition, on the basis of this study and polyphasic data obtained from previous work, it is proposed that the species Pseudoalteromonas sagamiensis should be reclassified as Algicola sagamiensis comb. nov. and that strain B-10-31(T) (=DSM 14643(T)=JCM 11461(T)) be designated the type strain.

Chryseobacterium caeni sp. nov., isolated from bioreactor sludge.

A Gram-negative, non-spore-forming, yellow-pigmented bacterium, strain N4(T), was isolated from a nickel-complexed cyanide-degrading bioreactor and subjected to a polyphasic taxonomic study. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain N4(T) is affiliated to the genus Chryseobacterium of the family Flavobacteriaceae. The levels of 16S rRNA gene sequence similarity between strain N4(T) and the type strains of all known Chryseobacterium species were 93.2-95.8 %, suggesting that strain N4(T) represents a novel species within the genus Chryseobacterium. The strain contained iso-C(15 : 0) and summed feature 4 as the major fatty acids and menaquinone MK-6 as the predominant respiratory quinone. The G+C content of the genomic DNA was 38.2 mol%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain N4(T) represents a novel species of the genus Chryseobacterium, for which the name Chryseobacterium caeni sp. nov. is proposed. The type strain is N4(T) (=KCTC 12506(T)=CCBAU 10201(T)=DSM 17710(T)).

Oceanisphaera donghaensis sp. nov., a halophilic bacterium from the East Sea, Korea.

A taxonomic study was carried out on two isolates, strains BL1(T) and BL11, from marine sediment collected from the East Sea, Korea. Comparative 16S rRNA gene sequence studies showed that these isolates clearly affiliated with the Gammaproteobacteria. BL1T and BL11 were most closely related to Oceanisphaera litoralis KMM 3654T (97.6 and 97.7 % 16S rRNA gene sequence similarity, respectively). The level of 16S rRNA gene sequence similarity between strains BL1T and BL11 was 99.7 %. The two isolates were Gram-negative, aerobic, moderately halophilic, and grew in 0.5-8.0 % NaCl and at 4-42 degrees C. Strains BL1T and BL11 shared some physiological and biochemical properties with O. litoralis KMM 3654T, although they differed in that BL1T and BL11 were able to utilize ethanol, proline and alanine. The G+C contents of the genomic DNA of strains BL1T and BL11 were 56.6 and 57.1 mol%, respectively. Both strains possessed C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, C(16 : 0) and C(18 : 1)omega7c as the major fatty acids. DNA-DNA relatedness data indicated that strains BL1T and BL11 represent a genomic species that is separate from O. litoralis KMM 3654T. On the basis of polyphasic evidence, it is proposed that strain BL1T (=KCTC 12522T = DSM 17589T) represents the type strain of a novel species, Oceanisphaera donghaensis sp. nov.

Azonexus caeni sp. nov., a denitrifying bacterium isolated from sludge of a wastewater treatment plant.

A polyphasic taxonomic study was carried out to determine the taxonomic position of a newly isolated denitrifying bacterium, designated Slu-05T, which had been isolated from sludge from the main aerobic treatment tanks of a municipal sewage treatment plant. Phylogenetic analysis based on comparative 16S rRNA gene sequencing indicated that strain Slu-05T was closely related to Azonexus fungiphilus LMG 19178T (96.4 % sequence similarity), the sole species in the genus Azonexus. Strain Slu-05T comprised Gram-negative, motile, non-spore-forming and slightly curved rods. The predominant respiratory lipoquinone was Q-8. The major fatty acids were C16:1omega7c, C16:0, C18:1 isomers and C10:0 3-OH. The G+C content of the genomic DNA was 65.6 mol%. The results of DNA-DNA hybridization (15.6 %) together with phenotypic determination showed that strain Slu-5T could be distinguished from A. fungiphilus. Moreover, some phenotypic properties concerning enzyme activity, the substrates utilized as carbon sources and growth conditions distinguish strain Slu-5T from A. fungiphilus. On the basis of the results obtained in this study, Slu-05T (=DSM 17719T=KCTC 12530T=CCBAU 10199T) is the type strain of a novel species of Azonexus, for which the name Azonexus caeni sp. nov. is proposed.

Rhizobium daejeonense sp. nov. isolated from a cyanide treatment bioreactor.

A polyphasic study was carried out to determine the taxonomic position of two aerobic, cyanide-degrading bacterial strains, designated L61T and L22, which had been isolated from a bioreactor for the treatment of nickel-complexed cyanide. The two isolates exhibited almost identical taxonomic characteristics. Phylogenetic analysis inferred from comparative 16S rRNA gene sequences indicated that the isolates fall in a sublineage of the genus Rhizobium comprising the type strains of Rhizobium giardinii, Rhizobium radiobacter, Rhizobium rubi, Rhizobium larrymoorei, Rhizobium vitis, Rhizobium undicola, Rhizobium loessense, Rhizobium galegae and Rhizobium huautlense. Cells of the two isolates are Gram-negative, aerobic, motile and non-spore-forming rods (0.6-0.7x1.1-1.3 microm), with peritrichous flagella. The DNA G+C content is 60.1-60.9 mol%. Cellular fatty acids are C(16 : 0) (2.2-3.3 %), C(18 : 0) (2.1-3.2 %), C(19 : 0) cyclo omega8c (9.9-16.8 %), C(20 : 3)omega6,9,12c (2.7-3.3 %), summed feature 3 (7.2-7.7 %) and summed feature 7 (67.8-73.7 %). The strains formed nodules on a legume plant, Medicago sativa. A nifH gene encoding denitrogenase reductase, the key component of the nitrogenase enzyme complex, was detected in L61T by PCR amplification by using a nifH-specific primer system. Strains L61T and L22 were distinguished from the type strains of recognized Rhizobium species in the same sublineage based on low DNA-DNA hybridization values (2-4 %) and/or a 16S rRNA gene sequence similarity value of less than 96 %. Moreover, some phenotypic properties with respect to substrate utilization as a carbon or nitrogen source, antibiotic resistance and growth conditions could be used to discriminate L61T and L22 from Rhizobium species in the same sublineage. Based on the results obtained in this study, L61T and L22 are considered to be representatives of a novel species of Rhizobium, for which the name Rhizobium daejeonense sp. nov. is proposed. The type strain is L61T (=KCTC 12121T=IAM 15042T=CCBAU 10050T).