• International Journal of Oral Biology
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• 제46권3호
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• pp.140-145
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• 2021

This study aimed to develop Lautropia mirabilis-specific quantitative real-time polymerase chain reaction (qPCR) primers based on the sequence of DNA-directed RNA polymerase subunit beta gene. The PrimerSelect program was used in designing of the qPCR primers, RTLam-F4 and RTLam-R3. The specificity of the qPCR primers were performed by conventional PCR with 37 strains of 37 oral bacterial species, including L. mirabilis. The sensitivity of the primers was determined by qPCR with the serial dilution of purified genomic DNA of L. mirabilis KCOM 3484, ranged from 4 ng to 4 fg. The data showed that the qPCR primers could detect only L. mirabilis strains and as little as 40 fg of genome DNA of L. mirabilis KCOM 3484. These results indicate that this qPCR primer pair (RTLam-F4/RTLam-R3) may be useful for species-specific detection of L. mirabilis in epidemiological studies of oral bacterial infectious diseases such as periodontal disease.

• Journal of Species Research
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• 제10권2호
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• pp.142-153
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• 2021

Algal genomics approaches provide a massive number of genome/transcriptome sequences and reveal the evolutionary history vis-à-vis primary and serial endosymbiosis events that contributed to the biodiversity of photosynthetic eukaryotes in the eukaryote tree of life. In particular, phylogenomic methods using several hundred or thousands of genes have provided new insights into algal taxonomy and systematics. Using this method, many novel insights into algal species diversity and systematics occurred, leading to taxonomic revisions. In addition, horizontal gene transfers (HGTs) of functional genes have been identified in algal genomes that played essential roles in environmental adaptation and genomic diversification. Finally, algal genomics data can be used to address the pangenome, including core genes shared among all isolates and partially shared strain-specific genes. However, some aspects of the pangenome concept (genome variability of intraspecies level) conflict with population genomics concepts, and the issue is closely related to defining species boundaries using genome variability. This review suggests a desirable future direction to merge algal pangenomics and population genomics beyond traditional molecular phylogeny and taxonomy.

• 한국균학회지
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• 제51권4호
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• pp.361-371
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• 2023

The study was undertaken to isolate and characterize wild yeast strains from soil samples collected in Seoul, Korea. Among the 19 yeast strains obtained, 17 were previously recorded species. The remaining two strains, Wickerhamomyces sp. GW1-4 and Archaeorhizomyces sp. YB4-103 were new species candidates. The genomic and microbiological characteristics of GW1-4 and YB4-103 were investigated. Phylogenetic analyses based on the 26S rRNA gene sequences and internal transcribed sequences, GW1-4 and YB4-103, represent a distinct lineage within the family Phaffomycetaceae and Archaeorhizomycetaceae, respectively. The GW1-4 and YB4-103 strains had the highest sequence homology with Wickerhamomyces xylosivorus NBRC 111553^T (88.97%) and Archaeorhizomyces finlayi CBS 128710^T (87.55%), respectively.

• 한국발생생물학회지:발생과생식
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• 제19권3호
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• pp.163-166
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• 2015

Genomic DNAs were extracted from the muscle of twenty-one specimens of three eel species collected in Anguilla japonica (AJ), Muraenesox cinereus (MC) and Conger myriaster (CM) from the Yellow Sea, respectively. In the present study, 7 oligonucleotides primers generated 191 specific loci in the AJ species, 226 in the (MC) species and 181 in the CM species, respectively. The primer BION-02 generated the most loci (a total of 83), with an average of 11.86 in the AJ species. The specific loci generated by oligonucleotides primers exhibited inter-individual-specific characteristics, thus revealing DNA polymorphisms. With regard to average bandsharing value (BS) results, individuals from Conger myriaster species (0.808) exhibited higher bandsharing values than did individuals from Muraenesox cinereus species (0.729) (P<0.05). The longest genetic distance (0.430) displaying significant molecular difference was also between individual no. 01 within Anguilla japonica eel species and individual no. 04 within Anguilla japonica species. In this study, the dendrogram resulted from reliable seven oligonucleotides primers, indicating three genetic clusters composed of group I (ANGUILLA 01~ANGUILLA 07), group II (MURAENESOX 08~MURAENESOX 14) and group III (CONGER 15~CONGER 21). The existence of species differentiation and DNA polymorphisms among three eel species were detected by PCR analysis. As mentioned above, a dendrogram revealed close relationships between individual identities within three eel species. High levels of a significant genetic distance among three eel species showed this PCR approach is one of the most suitable tools for individuals and/or species biological DNA studies.

• 한국물환경학회지
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• 제37권5호
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• pp.381-397
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• 2021

Off-flavor materials (geosmin and 2-methylisoborneol (2-MIB)) produced by microorganisms, such as, cyanobacteria and actinomycetes, cause freshwater use problems worldwide. Due to unpleasant taste and odor, these microorganisms have raised issues especially in drinking water resources. Recently, there has been increasing concern about 2-MIB and causal cyanobacteria, namely, Pseudanabaena, in Korea. However, material production and ecological dynamics remain largely unexplored. This study reviewed the distribution of Pseudanabaena, its species diversity, and the research trend of molecular ecology related to 2-MIB production in Korea. Based on published literature, we found that seven species of Pseudanabaena which include P. mucicola, P. limnetica, P. redekei, P. catenata, P. galeata, P. yagii, and P. cinerea appeared to occur in a variety of Korean water systems. All of these Pseudanabaena species were found in the North-Han River system (Lakes Soyang, Chuncheon, Uiam, and Paldang). Some of these species were also detected in other watersheds, but the precise species diversity was not identified. Species belonging to the Pseudanabaena genus are hard to classify through general microscopic alpha taxonomy, due to their very small cell size and similar morphological characters. Moreover, the potential of 2-MIB production cannot be detected by microscopic observation. Combining molecular ecological techniques, such as, environmental genomic materials (eDNA, eRNA) analyses to conventional methods could be useful to better understand the off-flavor material production and dynamics, thereby providing more efficient management strategies of freshwater systems.

• Parasites, Hosts and Diseases
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• 제62권2호
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• pp.180-192
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• 2024

Free-living amoebae (FLA) are found in diverse environments, such as soils, rivers, and seas. Hence, they can be used as bioindicators to assess the water quality based solely on their presence. In this study, we determined the presence of FLA in river water by filtering water samples collected from various sites and culturing the resulting filtrates. FLA were detected in all the water samples with varying quality grades (Grades I-V). The significant increase in the size of the amoebae population with the deterioration in the water quality. Monoxenic cultures of the amoebae were performed, and genomic DNAs were isolated, among which 18S rDNAs were sequenced to identify the amoeba species. Of the 12 species identified, 10 belonged to the Acanthamoeba genus; of the remaining 2 species, one was identified as Vannella croatica and the other as a species of Vermamoeba. Acanthamoeba was detected in samples with Grades I to VI quality, whereas the Vermamoeba species was present only in Grade I water. V. croatica was found exclusively in water with Grade II quality. Following morphological observations, genomic DNA was sequenced using 16S rDNA to determine whether the species of Acanthamoeba harbored endosymbionts. Most of the isolated Acanthamoeba contained endosymbionts, among which 4 species of endogenous bacteria were identified and examined using transmission electron microscopy. This study provides evidence that the distribution of amoebae other than Acanthamoeba may be associated with water quality. However, further confirmation will be required based on accurate water quality ratings and assessments using a more diverse range of FLA.

• The Plant Pathology Journal
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• 제22권3호
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• pp.215-221
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• 2006

Gibberella zeae (anamorph: Fusarium graminearum) is an important pathogen of cereal crops. This fungus produces a broad range of secondary metabolites, including polyketides such as aurofusarin (a red pigment) and zearalenone (an estrogenic mycotoxin), which are important mycological characteristics of this species. A screen of G. zeae insertional mutants, generated using a restriction enzyme-mediated integration (REMI) procedure, led to the isolation of a mutant (Z43R606) that produced neither aurofusarin nor zearalenone yet showed normal female fertility and virulence on host plants. Outcrossing analysis confirmed that both the albino and zearalenone-deficient mutations are linked to the insertional vector in Z43R606. Molecular characterization of Z43R606 revealed a deletion of at least 220 kb of the genome at the vector insertion site, including the gene clusters required for the biosynthesis of aurofusarin and zearalenone, respectively. A re-creation of the insertional event of Z43R606 in the wild-type strain demonstrated that the 220-kb deletion is responsible for the phenotypic changes in Z43R606 and that a large region of genomic DNA can be efficiently deleted in G. zeae by double homologous recombination. The results showed that 52 putative genes located in the deleted genomic region are not essential for phenotypes other than the production of both aurofusarin and zearalenone. This is the first report of the molecular characterization of a large genomic deletion in G. zeae mediated by the REMI procedure.

• Mycobiology
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• 제42권1호
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• pp.46-51
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• 2014

A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplified polymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealed differences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, was isolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of the degenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggesting that chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primers were designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76 and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp product in both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of the P2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in the normal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushrooms including 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.

• Bulletin of the Korean Chemical Society
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• 제27권5호
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• pp.699-704
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• 2006

Escherichia coli RNase P is a ribonucleoprotein composed of M1 RNA and C5 protein. Previously, analysis of RNA aptamers selected for C5 protein from a synthetic RNA library showed that C5 protein could bind various RNA molecules as an RNA binding protein. In this study, we searched cellular RNA motifs that could be recognized by C5 protein by a genomic SELEX approach. We found various C5 protein-binding RNA motifs derived from E. coli genomic sequences. Our results suggest that C5 protein interacts with various cellular RNA species in addition to M1 RNA.

• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.282-286
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• 2003

The purpose of this study was to isolate a specific DNA probe for the strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening mothod. The whole-genomic DNA of P. intermedia ATCC 25611 was isolated and purified. The HindIII-digested genomic DNAs from the strain were cloned by the random cloning method. To screen the strain-specific DNA probe, inverted dot blot hybridization tests were performed. In this assay, 20 ng of recombinant plasmids containing the HindIII-digested genomic DNA fragment were boiled and blotted onto a nylon membrane, and hybridized with digoxigenin-dUTP labeled genomic DNAs in a concentration of 100 ng/ml. Southern blot analysis was performed in order to confirm the results of the inverted dot blot hybridization tests. The data showed that a Pi34 probe (2.1 kbp; 1 out of 32 probes) was specific for P. intermedia strain ATCC 25611 and could be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.