• Parasites, Hosts and Diseases
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• v.60 no.4
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• pp.289-293
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• 2022

Blastocystis is a genus of unicellular heterokont parasites belonging to a group of organisms known as Stramenopiles, which includes algae, diatoms, and water molds. Blastocystis includes several species that habitat in the gastrointestinal tracts of organisms as diverse as humans, farm animals, birds, rodents, reptiles, amphibians, fish, and cockroaches. It is important to public health and distributed globally, but its prevalence in dogs in Korea has not been reported to date. Here, we collected 787 canine fecal samples and assessed Blastocystis infection by age, sex, region, season, and diarrhea symptoms. We determined Blastocystis subtypes using phylogenetic analyses based on 18S rRNA gene sequences. We identified, 10 Blastocystis positive samples (1.3%). A higher proportion of infected dogs was asymptomatic; however, infection rates did not significantly differ according to region, age, sex, and season. Phylogenetic analysis showed that the Blastocystis sp. identified belonged to 4 subtypes (STs), ST1, ST5, ST10, and ST14, thus revealed the genetic diversity of Blastocystis sp. in dogs Korean. This is first report on the presence of Blastocystis sp. in dogs Korean. This study revealed a lower infection rate than expected and differed from previous studies in STs. Further studies are warranted to observe the national infection status of Blastocystis in dogs and the genetic characteristics of this genus.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.32-39
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• 2016

Coastal plant species Tetragonia tetragonoides (Pall.) Kuntze native to the Dokdo islands was sampled and then 17 endophytic fungi were purely isolated based on morphological differences. The fungal isolates were characterized by their growth properties under NaCl concentration or pH gradient. Culture filtrates of the 17 fungal isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity. As the results, YH103 strain showed the highest plant growth-promoting activity among them. Phylogenetic analysis of the isolates was done by the maximum likelihood method based on partial internal transcribed spacer region (ITS region: contaning ITS1, 5.8S, and ITS2), beta-tubulin (BenA), and calmodulin (CaM) gene sequences. Chromatographic analysis of the strain YH103 culture filtrate showed the existence of gibberellins ($GA_4$, $GA_7$, $GA_8$, and $GA_{19}$). Finally, the strain YH103 was identified as Aspergillus tubingensis by microscopic observation and molecular analysis and, to our knowledge, this is the first report of GAs producing A. tubingensis.

• The Korea Journal of Herbology
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• v.27 no.6
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• pp.37-42
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• 2012

Objectives : Angelica Gigantis Radix is prescribed as the root of different Angelica species on the pharmacopoeia in Korea, Japan and China. Chemical components and their biological activities were also different according to their species. A study for the development of simple method to compare Angelica roots was needed. In order to classify them, the methods such as DNA sequencing analysis and taste sensor were applied to three Angelica species like Angelica gigas, Angelica acutiloba and Angelica sinensis. Methods : PCR amplification of intergenic transcribed spacer (ITS) region was performed using ITS1 and ITS4 primer from nine Angelica roots, and then nucleotide sequence was determined. Taste pattern of samples were measured using the taste-sensing system SA402B equipped with a sensing unit, which consists of artificial lipid membrane sensor probes of anionic bitterness, astringency, saltiness, umami, and cationic bitterness (C00, AE1, CT0, AAE, and AN0, respectively). Results : As a result of comparing the similarity of the ITS region sequences, A. sinensis was discriminated from the others (A. gigas and A. acutiloba). Equally this genetic result, A. gigas and A. acutiloba showed similar taste pattern as compared to A. sinensis. Sourness, bitterness, aftertaste of bitterness, astringency, and aftertaste of astringency of A. sinensis were significantly high as compared with A. gigas and A. acutiloba. In contrast, richness was significantly low. Conclusions : These taste pattern can be used as a way of comparison of Angelica species and this technic could be applied to establish a taste pattern marker for standardization of herbs in various purposes.

• Journal of Life Science
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• v.12 no.6
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• pp.821-834
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• 2002

In order to measure the inter- and intraspecific genetic divergences within the genus Alexandrium, the variations within the internal transcribed spacer (ITS1 and ITS2) regions and 5.85 ribosomal RNA gene of eight Alexandrium species were examined for 33 strains from diverse geographical locations by direct sequencing. Five isolates of A. tamarense (AT-2, AT-6, AT-10, AT-A and AT-B) from Jinhae Bay, Korea were found to be completely identical to a Japanese strain OFX151-A. The length of the amplified ITSI-5.85-ITS2 region varied from 481 nucleotides (in A. margalefi) to 528 nucleotides (in A. affine CU1-1). ITS1 and ITS2 nucleotide lengths were negatively correlated, whereas a positive correlation was found between their G＋C content. The degree of sequence divergence ranged from 0.3% (1 bp) to a maximum of 53% (305 Up). Pairwise sequence comparisons revealed a small degree of divergence between A. tamarense and A. Pundyense isolates (1.2 - 2.3% ＝ 6-12 bp), but a high degree of divergence between A. tamarense and A. catenella (19.8% ＝ 102 bp), and between A. catenella and A. Pundyense (19.7%). Although most nodes were weakly supported by bootstrap values, some types tend to form independent molecular groups. A. catenella isolates also formed an independent molecular sub-group, with relaticula strong bootstrap values (94% or 85% and 79% or 98%, respectively in PAUP and NJ trees). Interestingly, A. cohorticula and A. frateculus always clustered within the same sub-group, this result being supported by strong bootstrap values. Our results indicate that the ITS regions provide useful informations on hierarchical population genetic structure and a high phylogenetic resolution in intraspecific and interspecific Alexandrium population.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1366-1372
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• 2018

Objective: A disintegrin and metallopeptidase with thrombospondin motifs type 8 (ADAMTS8) is crucial for diverse physiological processes, such as inflammation, tissue morphogenesis, and tumorigenesis. The chicken ADAMTS8 (chADAMTS8) gene was differentially expressed in the kidney following exposure to different calcium concentrations, suggesting a pathological role of this protein in metabolic diseases. We aimed to examine the molecular characteristics of chADAMTS8 and analyze the gene-expression differences in response to toll-like receptor 3 (TLR3) stimulation. Methods: The ADAMTS8 mRNA and amino acid sequences of various species (chicken, duck, cow, mouse, rat, human, chimpanzee, pig, and horse) were retrieved from the Ensembl database and subjected to bioinformatics analyses. Reverse-transcription polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) experiments were performed with various chicken tissues and the chicken fibroblast DF-1 cell line, which was stimulated with polyinosinic-polycytidylic acid (poly[I:C]; a TLR3 ligand). Results: The chADAMTS8 gene was predicted to contain three thrombospondin type 1 (TSP1) domains, whose amino acid sequences shared homology among the different species, whereas sequences outside the TSP1 domains (especially the amino-terminal region) were very dif­ferent. Phylogenetic analysis revealed that chADAMTS8 is evolutionarily clustered in the same clade with that of the duck. chADAMTS8 mRNA was broadly expressed in chicken tissues, and the expression was significantly up-regulated in the DF-1 cells in response to poly(I:C) stimulation (p<0.05). These results showed that chADAMTS8 may be a target gene for TLR3 signaling. Conclusion: In this report, the genetic information of chADAMTS8 gene, its expression in chicken tissues, and chicken DF-1 cells under the stimulation of TLR3 were shown. The result suggests that chADAMTS8 expression may be induced by viral infection and correlated with TLR3-mediated signaling pathway. Further study of the function of chADAMTS8 during TLR3-dependent inflammation (which represents RNA viral infection) is needed and it will also be important to examine the molecular mechanisms during different regulation, depending on innate immune receptor activation.

• Korean Journal of Environment and Ecology
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• v.31 no.5
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• pp.461-471
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• 2017

Population genetic studies of 10 groups of Iksookimia pacifica were conducted to investigate the genetic diversity and population genetic structure across its known range in South Korea. Population DNA sequences of one mitochondrial gene (mtCOI) and three nuclear genes (IRBP, EGR2B, RAG1) were examined in samples collected from ten streams that flow into the East Sea. Both mitochondrial and nuclear sequences exhibited significant differentiation among populations except a few cases. The Bayesian analysis of the multi-locus genotypes inferred from the DNA sequences of nuclear genes clustered the individual fish largely into two geographical groups: a northern group (from Baebong stream to Cheonjin stream) and a southern group (Yangyangnamdae stream to Gangneungnamdae stream). Given that the streams flowing into the East Sea are geographically isolated water systems, such separation of genotypes can be interpreted by the geographical separation of common ancestors into north and south that had colonized South Korea. Since the initial geographical separation of the ancestral population by north and south, the ancestral groups seem to have experienced further differentiation into the current genetic clusters through the physical isolation of streams by the East Sea in each region. It is notable that many individuals in the Jasan stream formed a genetic cluster with those of Yangyangnamdae and Gangneungnamdae streams which are distant from each other. In addition, mitochondrial gene showed low genetic differentiation between some neighboring populations and very low level of genetic diversity in several populations. The present population genetic study will provide valuable information for the conservation and management of the Korean endemic fish species, I. paicifica.

• Journal of Animal Science and Technology
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• v.45 no.2
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• pp.169-182
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• 2003

To understand the structure and regulation of bovine ADRP (Adipocyte Differentiation Related Protein) gene, we have isolated the genomic clone of bovine ADRP and determined its sequence. A genomic Southern blot analysis confirmed that ADRP gene is present as a single copy in bovine genome and the ADRP gene spans 12 kb. Bovine ADRP genomic clone, HwADRPg-1, had 8 exons and 7 introns, and all splicing sites conformed to the GT/AG rule with the exon-intron boundaries located exactly. Analysis of the upstream 649 bp of the sequence of HwADRPg-1 showed that it does not contain any canonical TATAA boxes; however Sp1 binding sites and CAAT boxes are found. The promoter contained potential binding sites for AP-1, AP-2 and several putative transcription factor binding sites. The 5'-flanking region of HwADRPg-1 contained muscle specific transcription activator Myo G and C/EBP (CCAAT/ enhancer binding protein) recognizing site. These results suppose that the Myo G transcription activator regulate the transcription of bovine ADRP gene in muscular tissue and its transcriptional activity was triggered by degree of muscular development. Our results provide the necessary analysis for other flanking sequences are needed in addition to the proximal cis elements of this promoter to confer adipocyte differentiation-dependent or growth-dependent transcriptional control.

• Genomics & Informatics
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• v.21 no.3
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• pp.39.1-39.15
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• 2023

DNA barcoding without assessing reliability and validity causes taxonomic errors of species identification, which is responsible for disruptions of their conservation and aquaculture industry. Although DNA barcoding facilitates molecular identification and phylogenetic analysis of species, its availability in clariid catfish lineage remains uncertain. In this study, DNA barcoding was developed and validated for clariid catfish. 2,970 barcode sequences from mitochondrial cytochrome c oxidase I (COI) and cytochrome b (Cytb) genes and D-loop sequences were analyzed for 37 clariid catfish species. The highest intraspecific nearest neighbor distances were 85.47%, 98.03%, and 89.10% for COI, Cytb, and D-loop sequences, respectively. This suggests that the Cytb gene is the most appropriate for identifying clariid catfish and can serve as a standard region for DNA barcoding. A positive barcoding gap between interspecific and intraspecific sequence divergence was observed in the Cytb dataset but not in the COI and D-loop datasets. Intraspecific variation was typically less than 4.4%, whereas interspecific variation was generally more than 66.9%. However, a species complex was detected in walking catfish and significant intraspecific sequence divergence was observed in North African catfish. These findings suggest the need to focus on developing a DNA barcoding system for classifying clariid catfish properly and to validate its efficacy for a wider range of clariid catfish. With an enriched database of multiple sequences from a target species and its genus, species identification can be more accurate and biodiversity assessment of the species can be facilitated.

• The Korean Journal of Pesticide Science
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• v.17 no.4
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• pp.394-401
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• 2013

Trichoderma harzianum is one of rhizosphere fungus usually lives near the plant root regions in the soil. T. harzianum plays an important role in plant growth promotion and increases disease resistance against various plant pathogens on crops. In this study, the strain T. harzianum MPA167 was isolated from the barley rhizosphere soil in Suwon, Korea. Among 183 isolates, the strain T. harzianum MPA167 was selected as promising strain in which based on hyperparasitical activity against Phytophthora capsici and estimated disease control activity against P. capsici in the greenhouse conditions. The strain T. harzianum MPA167 was identified using 23s rDNA internal transcribed spacer(ITS) region sequences. MPA167 treatment ($1{\times}10^6$ spores/ml) showed greater disease suppression against Phytophthora blight of red-pepper caused by P. capsici in greenhouse compared with the water-treated control. Volatiles derived from T. harzianum MPA167 elicit growth promotion of tobacco and Arabidopsis seedlings in I-plate assay. In addition, T. harzianum MPA167 strain was also found to be effective for the growth promotion and induction of systemic resistance on red-papper plant. These results suggest that MPA167 might be used as one of the potential biocontrol agents.

• Horticultural Science & Technology
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• v.33 no.2
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• pp.242-250
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• 2015

Insertional mutagenesis induced by T-DNA or transposon tagging offers possibilities for analysis of gene function. However, its potential remains limited unless good methods for detecting the target locus are developed. We describe a PCR technique for efficient identification of DNA sequences adjacent to the inserted T-DNA in a higher plant, Chinese cabbage (Brassica rapa ssp. pekinensis). This strategy, which we named variable argument thermal asymmetric interlaced PCR (VA-TAIL PCR), was designed by modifying a single-step annealing-extension PCR by including a touch-up PCR protocol and using long gene-specific primers. Amplification efficiency of this PCR program was significantly increased by employing an autosegment extension method and linked sequence strategy in nested long gene-specific primers. For this technique, arbitrary degenerate (AD) primers specific to B. rapa were designed by analyzing the Integr8 proteome database. These primers showed higher accuracy and utility in the identification of flanking DNA sequences from individual transgenic Chinese cabbages in a large T-DNA inserted population. The VA-TAIL PCR method described in this study allows the identification of DNA regions flanking known DNA fragments. This method has potential biotechnological applications, being highly suitable for identification of target genomic loci in insertional mutagenesis screens.