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author:("Tago, kanao")
1.  A Fine-Scale Phylogenetic Analysis of Free-Living Burkholderia Species in Sugarcane Field Soil 
Microbes and Environments  2014;29(4):434-437.
The diversity and abundance of Burkholderia species in sugarcane field soils were investigated by a 16S rRNA gene-based approach using genus-specific primers. A total of 365,721 sequences generated by the Illumina MiSeq platform were assigned to the genus Burkholderia. Nearly 58% of these sequences were placed in a previously defined cluster, including stinkbug symbionts. Quantitative PCR analysis revealed a consistent number of 16S rRNA gene copies for Burkholderia species (107 g−1 soil) across the sampled fields. C/N, pH, and nitrate concentrations were important factors shaping the Burkholderia community structure; however, their impacts were not significant considering the overall genus size.
doi:10.1264/jsme2.ME14122
PMCID: PMC4262370
Burkholderia; Illumina sequencing; sugarcane field
2.  Bacterial population succession and adaptation affected by insecticide application and soil spraying history 
Although microbial communities have varying degrees of exposure to environmental stresses such as chemical pollution, little is known on how these communities respond to environmental disturbances and how past disturbance history affects these community-level responses. To comprehensively understand the effect of organophosphorus insecticide application on microbiota in soils with or without insecticide-spraying history, we investigated the microbial succession in response to the addition of fenitrothion [O,O-dimethyl O-(3-methyl-p-nitrophenyl) phosphorothioate, abbreviated as MEP] by culture-dependent experiments and deep sequencing of 16S rRNA genes. Despite similar microbial composition at the initial stage, microbial response to MEP application was remarkably different between soils with and without MEP-spraying history. MEP-degrading microbes more rapidly increased in the soils with MEP-spraying history, suggesting that MEP-degrading bacteria might already exist at a certain level and could quickly respond to MEP re-treatment in the soil. Culture-dependent and -independent evaluations revealed that MEP-degrading Burkholderia bacteria are predominant in soils after MEP application, limited members of which might play a pivotal role in MEP-degradation in soils. Notably, deep sequencing also revealed that some methylotrophs dramatically increased after MEP application, strongly suggesting that these bacteria play a role in the consumption and removal of methanol, a harmful derivative from MEP-degradation, for better growth of MEP-degrading bacteria. This comprehensive study demonstrated the succession and adaptation processes of microbial communities under MEP application, which were critically affected by past experience of insecticide-spraying.
doi:10.3389/fmicb.2014.00457
PMCID: PMC4148734  PMID: 25221549
fenitrothion; organophosphorus insecticide; soil microbes; deep sequencing; Burkholderia; methylotroph
3.  Effects of Elevated Carbon Dioxide, Elevated Temperature, and Rice Growth Stage on the Community Structure of Rice Root–Associated Bacteria 
Microbes and Environments  2014;29(2):184-190.
The effects of free-air carbon dioxide enrichment (FACE) and elevated soil and water temperature (warming) on the rice root–associated bacterial community were evaluated by clone library analysis of the 16S ribosomal RNA gene. Roots were sampled at the panicle initiation and ripening stages 41 and 92 days after transplanting (DAT), respectively. The relative abundances of the methanotrophs Methylosinus and Methylocystis were increased by warming and decreased by FACE at 92 DAT, which indicated that microbial methane (CH4) oxidation in rice roots may have been influenced by global warming. The relative abundance of Burkholderia kururiensis was increased by warming at 41 DAT and by FACE or warming at 92 DAT. The abundances of methanotrophs increased during rice growth, which was likely induced by an enhancement in the emission of CH4 from the paddy fields, suggesting that CH4 is one of the predominant factors affecting the structure of the microbial community in rice roots. Marked variations in the community structure were also observed during rice growth in other genera: Bradyrhizobium, Clostridium, and an unknown genus close to Epsilonproteobacteria were abundant at 92 DAT, whereas Achromobacter was abundant at 41 DAT. These results demonstrated that the community structures of rice root-associated bacteria were markedly affected by FACE, temperature, and the rice growth stage.
doi:10.1264/jsme2.ME14011
PMCID: PMC4103525  PMID: 24882221
carbon dioxide; methane; plant-associated bacteria; rice; warming
4.  Advantages of functional single-cell isolation method over standard agar plate dilution method as a tool for studying denitrifying bacteria in rice paddy soil 
AMB Express  2012;2:50.
We recently established a method for isolating functional single cells from environmental samples using a micromanipulator (Functional single-cell (FSC) isolation), and applied it to the study of denitrifying bacteria in rice paddy soil (Ashida et al. 2010. Appl Microbiol Biotechnol 85:1211–1217). To further examine the advantages and possible disadvantages of the FSC method, we isolated denitrifying bacteria from the same rice paddy soil sample using both FSC and standard agar plate dilution (APD) methods and compared in this study. The proportion of denitrifying bacteria in the total isolates was more than 6-fold larger with FSC isolation (57.1%) compared with the APD method (9.2%). Denitrifying bacteria belonging to Alphaproteobacteria and Bacilli were commonly isolated using both methods, whereas those belonging to Betaproteobacteria, which had been found to be active in the denitrification-inductive paddy soil, were isolated only with the FSC method. On the other hand, Actinobacteria were only isolated using the APD method. The mean potential denitrification activity of the FSC isolates was higher than that of the APD isolates. Overall, FSC isolation was confirmed to be an excellent method for studying denitrifying bacteria compared with the standard agar plate dilution method.
doi:10.1186/2191-0855-2-50
PMCID: PMC3488030  PMID: 22985609
16S rRNA gene; Denitrifying bacteria; Functional single-cell isolation; Phylogenetic analysis; Rice paddy soil
5.  Complete Genome Sequence of the Denitrifying and N2O-Reducing Bacterium Azoarcus sp. Strain KH32C 
Journal of Bacteriology  2012;194(5):1255.
We report the finished and annotated genome sequence of a denitrifying and N2O-reducing betaproteobacterium, Azoarcus sp. strain KH32C. The genome is composed of one chromosome and one megaplasmid and contains genes for plant-microbe interactions and the gene clusters for aromatic-compound degradations.
doi:10.1128/JB.06618-11
PMCID: PMC3294784  PMID: 22328754
6.  Complete Genome Sequence of the Denitrifying and N2O-Reducing Bacterium Pseudogulbenkiania sp. Strain NH8B 
Journal of Bacteriology  2011;193(22):6395-6396.
Pseudogulbenkiania sp. strain NH8B is a Neisseriales bacterium isolated from an agricultural field. This strain has strong denitrification and N2O reduction activities. Here, we report the finished and annotated genome sequence of this organism.
doi:10.1128/JB.06127-11
PMCID: PMC3209224  PMID: 22038961
7.  Phyllosphere yeasts rapidly break down biodegradable plastics 
AMB Express  2011;1:44.
The use of biodegradable plastics can reduce the accumulation of environmentally persistent plastic wastes. The rate of degradation of biodegradable plastics depends on environmental conditions and is highly variable. Techniques for achieving more consistent degradation are needed. However, only a few microorganisms involved in the degradation process have been isolated so far from the environment. Here, we show that Pseudozyma spp. yeasts, which are common in the phyllosphere and are easily isolated from plant surfaces, displayed strong degradation activity on films made from poly-butylene succinate or poly-butylene succinate-co-adipate. Strains of P. antarctica isolated from leaves and husks of paddy rice displayed strong degradation activity on these films at 30°C. The type strain, P. antarctica JCM 10317, and Pseudozyma spp. strains from phyllosphere secreted a biodegradable plastic-degrading enzyme with a molecular mass of about 22 kDa. Reliable source of biodegradable plastic-degrading microorganisms are now in our hands.
doi:10.1186/2191-0855-1-44
PMCID: PMC3293741  PMID: 22126328
Pseudozyma; Biodegradable plastic; Phyllosphere; Yeast

Results 1-7 (7)