• Korean Journal of Soil Science and Fertilizer
• /
• v.44 no.1
• /
• pp.127-145
• /
• 2011

Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

• Journal of Korean Society of Environmental Engineers
• /
• v.32 no.4
• /
• pp.369-378
• /
• 2010

Microorganisms are key-role player for stabilization of landfill sites. In order to evaluate the availability of T-RFLP(Terminal Restriction Fragment Length Polymorphism) for monitoring microbial community variations during stabilization of landfill sites, the phylogenic diversity of microbial community in the leachate from 4 different full-scale landfills was characterized by T-RFLP based on bacterial 16S rDNA. Main population of microbial community analyzed by T-RFLP was significantly similar with that of microbial community analyzed by clone library analysis. The results of T-RFLP analysis for main population of microbial community in the leachate from landfills with different landfill structures, waste types and landfill ages showed apparently different microbial diversity and structures. Therefore, long-term monitoring of microbial community in leachate from landfill sites by using T-RFLP is expected to be available for evaluation of landfill stability.

• Korean Journal of Microbiology
• /
• v.55 no.3
• /
• pp.220-225
• /
• 2019

Studies based on microbial community analyses have increased in the recent decade since the development of next generation sequencing technology. Associations of gut microbiota with host's health are one of the major outcomes of microbial ecology filed. The major approach for microbial community analysis includes the sequencing of variable regions of 16S rRNA genes, which does not provide the information of bacterial activities. Here, we conducted RNA-based microbial community analysis and compared results obtained from DNA- and its cDNA-based microbial community analyses. Our results indicated that these two approaches differed in the ratio of Firmicutes and Bacteroidetes, known as an obesity indicator, as well as abundance of some key bacteria in gut metabolisms such as butyrate producers and probiotics strains. Therefore, cDNA-based microbial community may provide different insights regarding roles of gut microbiota compared to the previous studies where DNA-based microbial community analyses were performed.

• Korean Journal of Soil Science and Fertilizer
• /
• v.46 no.6
• /
• pp.474-481
• /
• 2013

Understanding the microbial community structure of agricultural soils is important for better soil management in order to improve soil quality. Phospholipid fatty acid analysis has been popularly used in determining the microbial community structure in different ecosystems. The microbial community structure of paddy soil under long-term fertilizer treatments was investigated after 45 years using PLFA analysis. Treatments were control (no fertilization, Con), compost (COM), NPK, NPK+compost (NPKC), PK, NK, and NP. Soil chemical properties were mainly affected by the addition of compost and inorganic P fertilizer. Total nitrogen and organic matter contents were significantly higher in treatments with compost while available $P_2O_5$ and exchangeable calcium were significantly higher in treatments with added inorganic P fertilizer. It was found that microbial communities were responsive to the different fertilizer treatments. PLFA results showed that the soils were dominated by gram-negative bacteria, followed by the actinomycetes, then gram-positive bacteria, and fungi. Principal component analysis of the soil chemical properties and PLFA composition proved to be a more reliable tool because it was more responsive to the changes in soil chemical properties.

• Korean Journal of Environmental Agriculture
• /
• v.38 no.3
• /
• pp.185-196
• /
• 2019

BACKGROUND: This study investigated the effects of rice genetically modified to be resistant against rice blast and rice bacterial blight on the soil microbial community. A comparative analysis of the effects of rice genetically modified rice choline kinase (OsCK1) gene for disease resistance (GM rice) and the Nakdong parental cultivar (non-GM rice) on the soil microbial community at each stage was conducted using rhizosphere soil of the OsCK1 and Nakdong rice. METHODS AND RESULTS: The soil chemistry at each growth stage and the bacterial and fungal population densities were analyzed. Soil DNA was extracted from the samples, and the microbial community structures of the two soils were analyzed by pyrosequencing. No significant differences were observed in the soil chemistry and microbial population density between the two soils. The taxonomic analysis showed that Chloroflexi, Proteobacteria, Firmicutes, Actinobacteria, and Acidobacteria were present in all soils as the major phyla. Although the source tracking analysis per phylogenetic rank revealed that there were differences in the bacteria between the GM and non-GM soil as well as among the cultivation stages, the GM and non-GM soil were grouped according to the growth stages in the UPGMA dendrogram analysis. CONCLUSION: The difference in bacterial distributions between Nakdong and OsCK1 rice soils at each phylogenetic level detected in microbial community analysis by pyrosequencing may be due to the genetic modification done on GM rice or due to heterogeneity of the soil environment. In order to clarify this, it is necessary to analyze changes in root exudates along with the expression of transgene. A more detailed study involving additional multilateral soil analyses is required.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.6
• /
• pp.765-770
• /
• 2015

Recent sequencing technology development has revolutionized fields of microbial ecology. MiSeq-based microbial community analysis allows us to sequence more than a few hundred samples at a time, which is far more cost-effective than pyrosequencing. The approach, however, has not been preferably used owing to computational difficulties of processing huge amounts of data as well as known Illumina-derived artefact problems with amplicon sequencing. The choice of assembly software to take advantage of paired-end sequencing and methods to remove Illumina artefacts sequences are discussed. The protocol we suggest not only removed erroneous reads, but also dramatically reduced computational workload, which allows even a typical desktop computer to process a huge amount of sequence data generated with Illumina sequencers. We also developed a Web interface (http://biotech.jejunu.ac.kr/ ~abl/16s/) that allows users to conduct fastq-merging and mothur batch creation. The study presented here should provide technical advantages and supports in applying MiSeq-based microbial community analysis.

• Environmental Engineering Research
• /
• v.14 no.1
• /
• pp.3-9
• /
• 2009

Microorganisms play an important role in the geochemical cycles, industry, environmental cleanup, and biotechnology among other fields. Given the high microbial diversity, identification of the microorganism is essential in understanding and managing the processes. One of the most popular and powerful method for microbial identification is comparative 16S rRNA gene analysis. Due to the highly conserved nature of this essential gene, sequencing and later comparison of it against known rRNA databases can provide assignment of the bacteria into the taxonomy, and the identity of its closest relatives. Isolation and sequencing of 16S rRNA genes directly from natural environments (either from DNA or RNA) can also be used to study the structure of the whole microbial community. Nowadays, novel sequencing technologies with massive outputs are giving researchers worldwide the chance to study the microbial world with a depth that was previously too expensive to achieve. In this article we describe commonly used research approaches for the study of individual microorganisms and microbial communities using the tools provided by Ribosomal Database Project website.

• KSBB Journal
• /
• v.28 no.5
• /
• pp.275-281
• /
• 2013

Soil microbial community analysis of farmland soil sprayed with lye in order to use fertilizer in Nigeria was performed. As a control, two kinds of soils not sprayed with lye, located in Eungo and Lagos with general practice in agriculture was selected. Soil sprayed with lye was pH 8.25 through alkalization reaction, while the other soil samples were pH 6.22 and 5.94. Substrate utilization and species diversity index of soil sprayed with lye were low than that of the other soils with the analysis of Biolog ecoplate. As a result of principal component analysis, the relationship between three samples was low. Microbial community analysis was performed by DGGE and most of them were soil uncultured bacterium. Especially, Uncultured Acidobacteria and Uncultured Methylocystis sp., which had been isolated from the rhizosphere of soybean grown in that site were discovered in the soil sprayed with lye.

• Korean Chemical Engineering Research
• /
• v.48 no.2
• /
• pp.147-156
• /
• 2010

There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

• Journal of Korean Society of Water and Wastewater
• /
• v.20 no.5
• /
• pp.685-690
• /
• 2006

The differences in microbial community structures between planktonic microorganism and biofilm in rivers were investigated using respiratory quinone profiles. The compositions of microbial quinone for 4 tributaries of the Kyongan Stream located in/flowing through Yongin City, Gyeonggi-Do were analyzed. Ubiquinone(UQ)-8, UQ-9, menaquinone(MK)-6 and Plastoquinone(PQ)-9 were observed in all samples of planktonic microorganism and biofilm for the sites investigated, Most planktonic microorganism and biofilm had UQ-8(15 to 30%) and PQ-9(over 30%) as the dominant quinone type. These results indicated that oxygenic phototrophic microbes(cyanobacteria and/or eukaryotic phytoplankton) and UQ-8 containing proteobacteria constituted major microbial populations in the river. The quinone concentration in the river waters tested, which reflects the concentration of planktonic microorganisms, increases with increasing DOC. Further research into this is required. The microbial diversities of planktonic microorganism and biofilm calculated based on the composition of all quinones were in the range from 4.2 to 7.5, which was lower than those for activated sludge(ranging from 11 to 14.8) and soils(ranging from 13.4 to 16.8). The use of quinone profile appears to be a useful tool for the analysis of microbial community structure in river.