• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.126-126
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• 2018

Many of researches have revealed that human intestinal microbiota is related to health. Several diseases like obesity, diabetes, and hypertension are affected by the microbiota directly and indirectly. So, interventions with food and drug have been tried to change a composition of the microbiota to better condition. However, few natural medicines have elucidated to date. To understand an influence on microbiota by plant materials including Glycyrrhizae Radix, the extract of medicines were administered to mice and the feces were collected before and after the administration. The feces were analyzed by terminal restriction fragment length polymorphism (T-RFLP). The changes in composition of mice gut microbiota were detected and analyzed. The data could be utilized to further study about biological activities of the plant medicines.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.563-566
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• 2012

To investigate the possibility of horizontal gene transfer between agricultural microorganisms and soil microorganisms in the environment, Bacillus subtilis KB producing iturin and the PGPR recombinant strain Pseudomonas fluorescens MX1 were used as model microorganisms. The soil samples of cucumber or tomato plants cultivated in pots and the greenhouse for a six month period were investigated by PCR, real-time PCR, Southern hybridization, and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Our data from Southern blotting and T-RFLP patterns suggest that the model bacteria do not give significant impacts on the other bacteria in the pots and greenhouse during cultivation.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.103-110
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• 2006

T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in Zoysia japonica soil treated with liquid fertilizer containing amino acids(LFcAA) after spray with herbicide. The results of T-RFLP (terminal restriction fragment length poly-morphism) analysis using restriction enzyme Hae III showed that the T-RFs of various size appeared evenly in the 32 clones of KD3 and 38 clones of KD4 respectively that had been treated with liquid fertilizer containing amino acid(LFcAA) compared to 23 clones of KD2 hat had not been treated with LFcAA. The microbial com- munity structure in KD2 appeared less diverse than those in KD3 and KD4. Analysis of partial sequences for 110 clones from KDI (control), KD2 (non-treated), KD3 (LFcAA 1X), KD4 (LFcAA 2X), respectively, revealed that most bacteria were related with uncultured bacteria in a 16S rDNA sequence similarity range of 91-99% through blast search. Otherwise, the other clones were members of proteobacteria, Acidobacteria, Act-inobacteria, Sphingobacteria and Planctomyces groups. Especially in KD4, members of Alpha Proteobacteria, Rhizobiales, Sphigomonadales, Caulobacterales, Gamma Proteobacteria, the genus Pseudomonas, Betapro-teobacteria, Nitrosomonadales and genus Nitrosospira appeared to be dominant. In addition, Acidobacteria group, Actinobacteria group, Planctomycetacia and Sphingobacteria were also shown. The microbial com-munity structure in Z. japonica soil sprayed with herbicide was affected by LFcAA.

• Tuberculosis and Respiratory Diseases
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• v.79 no.3
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• pp.165-178
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• 2016

Background: Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid $na{\ddot{i}}ve$ asthmatics. Methods: Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-$na{\ddot{i}}ve$ asthma patients were analyzed through T-RFLP analysis. Results: In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae, and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. Conclusion: Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

• Journal of Ecology and Environment
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• v.41 no.3
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• pp.99-106
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• 2017

Background: Plants over-expressing Arabidopsis ABF3 (abscisic acid-responsive element-binding factor 3) have enhanced tolerance to various environmental stresses, especially drought. Using terminal restriction fragment length polymorphism (T-RFLP) analysis, we compared the rhizosphere-associated structures of microbial communities for transgenic potato containing this gene and conventional "Jopoong" plants. Results: During a 2-year field experiment, fungal richness, evenness, and diversity varied by year, increasing in 2010 when a moderate water deficit occurred. By contrast, the bacterial richness decreased in 2010 while evenness and diversity were similar in both years. No significant difference was observed in any indices for either sampling time or plant line. Although the composition of the microbial communities (defined as T-RF profiles) changed according to year and sampling time, differences were not significant between the transgenic and control plants. Conclusions: The results in this study suggest that the insertion of ABF3 into potato has no detectable (by current T-RFLP technique) effects on rhizosphere communities, and that any possible influences, if any, can be masked by seasonal or yearly variations.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.15-21
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• 2003

Two microbial communities of activated sludge in the same municipal wastewater, but treated with different systems, were studied and compared using molecular microbiological approaches. The bacterial 16S rDNA sequences from 124 clones were analyzed, however, the majority of them were not closely related to any known species, and found to belong to 8 different phylogenetic groups and 3 different unidentified groups. The relative frequencies of each group were similar between the two microbial communities. Fingerprinting using terminal restriction fragment length polymorphism (T-RFLP) showed that the putative Nitrospira-related populations were more diverse and quantitatively higher in the KNR process system than in the other system using a conventional activated sludge process. The relationship between the bacterial community composition and the higher removal efficiency of nitrogen and phosphorus in the KNR process is discussed.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.627-633
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• 1996

To analyze the genomic diversity of transmissible gastroenteritis virus (TGEV), the N-terminal half of the spike (S) glycoprotein gene and nonstructural protein gene (open reading frames 3 and 3-1) were amplified by reverse transcriptase reaction and polymerase chain reaction (RT-PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. In this study, TGEV Miller (M6) and Purdue (P115) strains were used as reference strains, and two vaccine strains (MSV and STC3) and four Korea isolates (P44, VRI-WP, VRI-41, and VRI-48) were analyzed. All TGEV strains were amplified with three TGEV primer pairs. Although there was some exception in RFLP analysis, this method differentiated TGEV strains into following groups : Miller group (M6 and MSV), Purdue group (PUS, STC3, P44, VRI-WP, VRI-41, and VRI-48). Using Sau3AI and SspI, VRI-48 was differentiated from the Miller and Purdue type viruses. The RT/PCR in conjuction with RFLP analysis was a rapid and valuable tool for differentiating several strains of TGEV. This study revealed the occurences of distinct difference in genome of TGEV strains.

• The Plant Pathology Journal
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• v.19 no.1
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• pp.57-63
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• 2003

An insertion sequence identified as a solo long terminal repeat (LTR) of a new rice copia-like retrotransposon was detected in the ORE of the Pi-b gene from the rice cv. Nipponbare, and was designated as Osr1. Osr1 consists of a 6386 bp nucleotide sequence including 965 bp LTRs on both ends with an 82％ nucleotide sequence identity to the wheat Tarl retrotransposon on reverse transcriptase. Nucleotide divergence was noted among the individual LTRs, as well as the coding region of Osr1. Various restriction fragment length polymorphism (RFLP) of LTR were detected in indica cultivars, whereas, only a few could be detected in the japonica cultivars. The population of Osr1 is lower in the wild-type rice compared with that in the domesticated cultivars. The insertion of LTR sequence in the h-b gene in the susceptible cultivar suggested that retro-tyansposon-mediated insertional mutation might play an important role in the resistance breakdown, as well as in the evolution of resistance genes in rice.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.76-81
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• 2012

Removal of methane, benzene and toluene was evaluated in a lab-scale biocover packed with a soil mixture of forest soil and earthworm cast (75:25 weight ratio). The bacterial community in the biocover was characterized using quantitative real-time PCR and terminal restriction fragment length polymorphism. Methane was removed at the upper layer of the biocover (-0.1 ~ -0.4 m), where the oxygen concentration was remarkably lower. The average removal efficiencies for methane and benzene/toluene were 90% and 99%, respectively. The pmoA gene copy numbers, responsible for methane oxidation, in the upper layer were higher than those in the lower layer. While type I methanotrohs dominated the lower layer, type II methanotrophs, such as Methylocystis and Methylosinus, were noted to be predominant in the upper layer. Benzene and toluene were removed from the lower layer (-0.6 ~ -0.9 m) as well as the upper layer. Moreover, the tmoA gene copy number, responsible for benzene/toluene oxidation, seen in the upper layer was not significantly different from those seen in the lower layer. These results suggest that a biocover packed with a soil and earthworm cast mixture is a promising method which could be utilized for the control of methane and volatile organic compounds such as benzene and toluene.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1207-1215
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• 2008

To investigate the occurrence and species diversity of mycobacteria in waters, surface water samples were collected monthly from the Han River and tap water samples at the terminal sites of the distribution system. Mycobacteria in each water sample were isolated by decontamination using cetylpyridinium chloride (CPC) and cultivation on Middlebrook 7H10 agar, and then identified by polymerase chain reaction-restriction fragment length polymorphism analysis (PRA) and sequencing of the 65-kDa heat-shock protein gene (hsp65 gene). Mycobacteria were detected in 59% of the surface water samples and 26% of the tap water samples. Over half of the 158 isolates could not be identified by hsp65 PRA and gene sequencing, and several identification discrepancies were observed between the two methods. The most frequently isolated species was Mycobacterium gordonae in surface water and M. lentiflavum in tap water. M. avium complex (MAC), the most important pathogen among environmental mycobacteria, was detected in the surface water samples but not found in the tap water samples. The result demonstrated that water is an important environmental source of mycobacteria and the combined application of hsp65 PRA and sequencing was more reliable than hsp65 PRA alone to accurately identify mycobacteria present in water.