• Research in Plant Disease
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• v.10 no.1
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• pp.63-68
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• 2004

Twenty-three strains of Serratia sp., isolated from surface-sterilized rice roots collected in Chonbuk and Chungnam province, were identified and characterized. They were Gram-negative, rod shaped and red pigmented typically and their endophytism was confirmed by inoculation and reisolation of the strains in planta. Their antifungal activity against 4 rice pathogenic fungi was compared and ranged from 62.4 to 85.2％ against Rhizoctonia solani and 68.0 to 88.5％ against Pyricularia grisea. Among the 23 strains tested, strain Rsm220 showed the strongest inhibition activity against 4 pathogenic fungi. The strain was, therefore, selected as a biocontrol candidate for both the pathogens and its bacteriological characteristics and 165 rDNA sequences were analyzed. Phenotypic and biochemical characteristics of the selected Rsm220 were highly related to the type strain of S. marcescens and 165 rDNA sequencing of Rsm220 showed a homology of 98.2％ to the type strain of S. marcescens. The strain Rsm220 was identified as S. marcescens and the inhibition result of this endophytic strain indicates that it is a potential biocontrol agent for R. solani and R grisea.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.740-747
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• 2006

Amplification by universal consensus sequences in pathogenic bacterial DNA would allow rapid identification of pathogenic bacteria, and amplification of genus-specific and species-specific sequences of pathogenic bacterial DNA might be used for genotyping at the genus and species levels. For design of probes for molecular diagnostics, several tools are available as stand-alone programs or as Web application. However, since most programs can design only a few probe sets at one time, they are not suitable for large-scale and automatic probes design. Therefore, for high-throughput design of specific probes in diagnostic array development, an automated design tool is necessary. Thus, we developed a Web-based automatic system for design of genus-specific and species-specific probes for pathogen detection. The system is available at http://www.miprobe.com.

• Microbiology and Biotechnology Letters
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• v.43 no.1
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• pp.65-78
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• 2015

The bacterial diversity of the rhizosphere soil of S. japonica native to Suncheon bay was analyzed. Ninety two strains showing different morphological characteristics were isolated from the soils around the community of S. japonica. Bacterial diversity and distributions were studied by phylogenetic analysis of the partial 16S rDNA sequences. Ninety two strains were partially sequenced and analyzed phylogenetically. These strains were composed of 5 phyla firmicutes (56.5%), gamma-proteobacteria (29.3%), alpha-proteobacteria (5.4%), actinobacteria (5.4%), bacteroidetes (3.3%) and Shannon’s diversity index (H') were different from each of sampling sites (1.675, 1.924 and 2.04). Eleven isolates were presumed to be novel species candidates based on similarity analysis of the 16s rRNA gene sequences. Overall, Firmicutes and gamma-proteobacteria of the rhizosphere soil of S. japonica showed a high diversity.

• Journal of Microbiology and Biotechnology
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• v.16 no.3
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• pp.431-437
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• 2006

Aero-tolerant microorganisms were isolated from healthy Koreans over the age of 95 years. The microorganisms were then identified based on their morphological and biochemical characteristics and 16S rDNA sequences. The growth properties of the isolated strains were investigated in kimchi. The characteristics of kimchi containing the microorganisms were studied microscopically, physicochemically, and organoleptically. Among 7 aero-tolerant strains, a strain with a 16S rDNA sequence exhibiting 99% homology with Bifidobacterim animalis strain B83 was selected and named B. animalis DY-64. The new strain showed a better acid resistance and salt resistance (p<0.05) than B. animalis ATCC 25527. After 15 days of fermentation in kimchi, the viability of B. animalis DY-64 was about 10%, and the kimchi had a better overall edible quality than conventional kimchi. Thus, it was found that the application of B. animalis DY-64 to kimchi preparation produced a good overall edible quality.

• Parasites, Hosts and Diseases
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• v.43 no.1 s.133
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• pp.7-13
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• 2005

The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of $0.2-1.1\%$ with each other, but more than $2\%$ difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group lion the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1816-1821
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• 2003

Reliable PCR based identification of lactobacilli has been described utilizing the sequence of 16S-23S rRNA intergenic spacer region. Those sequence comparisons showed a high degree of difference in homology among the strains of L. rhamnosus, L. casei, L. acidophilus and L. helveticus whose 16S-23S rRNA intergenic small SR's sizes were 222 bp, 222 bp, 206 bp and 216 bp respectively. The sequence of 16S-23S rRNA intergenic spacer region of L. rhamnosus ATCC 53103 revealed the close relatedness to those of L. casei strains by the homology ranges from 95.4% to 97.2%. 16S-23S rRNA intergenic spacer region nucleotide sequence of L. acidophilus showed some distant relatedness with L. rhamnosus ATCC 53103 with the homology ranges from 40.3% to 41.8% and that with L. helveticus was shown to be 30% of homology, which exists at the most distant phylogenetic relatedness. The identification of species and strain of lactobacilli was possible on the basis of these results. The common sequences among the 17 strains were CTAAGGAA located in the initiating position of the DNA and some discrepancies were found between the same strains based on these results.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.398-406
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• 2015

Culture-dependent ARDRA and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Halichondria panicea collected from Jeju Island. A total of 120 bacterial strains associated with the sponge were cultivated using modified Zobell and Marine agar media. PCR amplicons of the 16S rRNA gene from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rRNA gene sequences derived from ARDRA patterns showed more than 96% similarities compared with known bacterial species, and the isolates belonged to four classes, Alphaproteobacteria, Gammaproteobacteria, Bacteroidetes, and Firmicutes, of which Alphaproteobacteria was dominant. DGGE fingerprinting of 16S rRNA genes amplified from the sponge-derived total gDNA showed 14 DGGE bands, and their sequences showed 100% similarities compared with the sequences available in GenBank. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of seven classes, including Alphaproteobacteria, Gammaproteobacteria, Acidobacteria, Actinobacteira, Bacteroidetes, Cyanobacteria, and Chloroflexi. According to both the ARDRA and DGGE methods, three classes, Alphaproteobacteria, Gammaproteobacteria, and Bacteroidetes, were commonly found in H. panicea. However, overall bacterial community in the sponge differed depending on the analysis methods. Sponge showed more various bacterial community structures in culture independent method than in culture-dependent method.

• Journal of Microbiology
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• v.41 no.3
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• pp.183-188
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• 2003

We isolated and cultured bacteria that inhabited marine biofilms, and identified them by phylogenetic analysis using 16S rDNA sequences. In the marine environment, biofilms cover most subtidal and intertidal solid surfaces such as rocks, ships, loops, marine animals, and algae. The bacteria in most biofilms are embedded in extracellular polymeric substances that comprise mainly of exopolysaccharides. The exopolysaccharides are excreted from multiple bacterial species; therefore, biofilms are a good source for screening exopolysaccharide-producing bacteria. Thirty-one strains were cultured, and a total of 17 unique strains were identified. Phylogenetic analysis using 16S rDNA sequences indicated that the 17 strains belonged to ${\alpha}$-Proteobacteria (Ochrobactrum anthropi, Paracoccus carotinifaciens); ${\gamma}$-Proteobacteria (Pseudoalteromonas agarovorans, P. piscicida, Pseudomonas aeruginosa, Shewanella baltica, Vibrio parahaemolyticus, V. pomeroyi); CFB group bacteria (Cytophaga latercula, Tenacibaculum mesophilum); high GC, Gram-positive bacteria (Arthrobacter nicotianae, Brevibacterium casei, B. epidermidis, Tsukamurella inchonensis); and low GC, Gram-positive bacteria (Bacillus macroides, Staphylococcus haemolyticus, S. warneri).

• Genomics & Informatics
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• v.10 no.4
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• pp.249-255
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• 2012

In this study, fosmid cloning strategies were used to assess the microbial populations in water from the International Space Station (ISS) drinking water system (henceforth referred to as Prebiocide and Tank A water samples). The goals of this study were: to compare the sensitivity of the fosmid cloning strategy with that of traditional culture-based and 16S rRNA-based approaches and to detect the widest possible spectrum of microbial populations during the water purification process. Initially, microbes could not be cultivated, and conventional PCR failed to amplify 16S rDNA fragments from these low biomass samples. Therefore, randomly primed rolling-circle amplification was used to amplify any DNA that might be present in the samples, followed by size selection by using pulsed-field gel electrophoresis. The amplified high-molecular- weight DNA from both samples was cloned into fosmid vectors. Several hundred clones were randomly selected for sequencing, followed by Blastn/Blastx searches. Sequences encoding specific genes from Burkholderia, a species abundant in the soil and groundwater, were found in both samples. Bradyrhizobium and Mesorhizobium, which belong to rhizobia, a large community of nitrogen fixers often found in association with plant roots, were present in the Prebiocide samples. Ralstonia, which is prevalent in soils with a high heavy metal content, was detected in the Tank A samples. The detection of many unidentified sequences suggests the presence of potentially novel microbial fingerprints. The bacterial diversity detected in this pilot study using a fosmid vector approach was higher than that detected by conventional 16S rRNA gene sequencing.

• Journal of fish pathology
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• v.17 no.3
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• pp.159-169
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• 2004

Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.