• Korean Journal of Veterinary Service
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• v.39 no.4
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• pp.259-266
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• 2016

Porcine epidemic diarrhea virus (PEDV) causes an acute and lethal watery diarrhea in piglets that is great economic losses to the swine country worldwide. The spike (S) glycoprotein is an important determinant for PEDV biological properties. In the present study, we determined the full-length S gene sequences of five Chung-nam PEDV field isolates collected in 2016. The S gene was amplified by RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. 5 field strains share 98.5%~99.9% homologies with each other at the nucleotide sequence level and 96.7%~99.9% homologies with each other at the amino acids sequence level. Most field strains have nucleotide insertions, deletions and mutation regions, and show lower homologies (93.1~93.8%) with classical and vaccine strains, however higher homologies (99.1%~99.5%) with US PEDV isolates in 2013. By phylogenetic tree analysis based on nucleotide sequence, five PEDV field isolates were clustered into Genogroup 2b but differ genetically from the vaccine strains (SM-98 and DR-13).

• Korean Journal of Veterinary Research
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• v.56 no.1
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• pp.23-28
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• 2016

Porcine epidemic diarrhea virus (PEDV), a porcine enteropathogenic coronavirus, causes lethal watery diarrhea in piglets, resulting in large economic losses because of high mortality. In November 2013, PEDV reemerged in Korea, and these outbreaks have since continuously occurred. In the present study, we determined the full-length nucleocapsid (N) gene sequences of three Korean PEDV field isolates collected in 2014-2015. Sequence and phylogenetic analysis of N genes revealed that recent prevalent Korean PEDV isolates were very closely related to the US PEDV isolates in 2013. Interestingly, the phylogenetic tree based on the nucleotide sequencing of the PEDV N gene was similar to the tree topology of the PEDV complete genomes. Therefore, our data provide a better understanding of the genetic diversity and contribute to the accurate diagnosis and development of vaccines against PEDV.

• The Plant Pathology Journal
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• v.34 no.6
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• pp.514-531
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• 2018

Tomato spotted wilt virus (TSWV; Genus Orthotospovirus: Family Tospoviridae) is one of the most destructive viruses affecting a wide range of horticultural crops on a worldwide basis. In 2015 and 2016, 171 leaf and fruit samples from tomato (Solanum lycopersicum) plants with viral symptoms were collected from the fields in various regions of Iran. ELISA test revealed that the samples were infected by TSWV. The results of RT-PCR showed that the expected DNA fragments of about 819 bp in length were amplified using a pair of universal primer corresponding to the RNA polymerase gene and DNA fragments of ca 777 bp and 724 bp in length were amplified using specific primers that have been designed based on the nucleocapsid (N) and non-structural (NSs) genes, respectively. The amplified fragments were cloned into pTG19-T and sequenced. Sequence comparisons with those available in the GenBank showed that the sequences belong to TSWV. The high nucleotide identity and similarities of new sequences based on the L, N, and NSs genes showed that minor evolutionary differences exist amongst the isolates. The phylogenetic tree grouped all isolates six clades based on N and NSs genes. Phylogenetic analysis showed that the Iranian isolates were composed a new distinct clade based on a part of polymerase, N and NSs genes. To our knowledge, this is the first detailed study on molecular characterization and genetic diversity of TSWV isolates from tomato in Iran that could be known as new clade of TSWV isolates.

• Journal of Veterinary Science
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• v.23 no.5
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• pp.73.1-73.8
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• 2022

Background: Lumpy skin disease (LSD), a disease transmitted by direct and indirect contact with infected cattle, is caused by the Lumpy skin disease virus (LSDV). The disease affects cattle herds in Africa, Europe, and Asia. The clinical signs of LSD range from mild to the appearance of nodules and lesions in the skin leading to severe symptoms that are sometimes fatal with significant livestock economic losses. Objectives: This study aimed to characterize LSDV strains in the blood of infected cattle in Thailand based on the GPCR gene and determine the phylogenetic relationship of LSDV Thailand isolates with published sequences available in the database. Methods: In total, the blood samples of 120 cattle were collected from different farms in four provinces in the northeastern part of Thailand, and the occurrence of LSDV was examined by PCR based on the P32 antigen gene. The genetic diversity of LSDV based on the GPCR gene was analyzed. Results: Polymerase chain reaction assays based on the P32 antigen gene showed that 4.17% (5/120) were positive for LSDV. All positive blood samples were amplified successfully for the GPCR gene. Phylogenetic analysis showed that LSDV Thailand isolates clustered together with LSDVs from China and Russia. Conclusions: The LSD outbreak in Thailand was confirmed, and a phylogenetic tree was constructed to infer the branching pattern of the GPCR gene from the presence of LSDV in Thailand. This is the first report on the molecular characterization of LSDV in cattle in Thailand.

• Journal of Plant Biotechnology
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• v.49 no.4
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• pp.271-291
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• 2022

Pectin methylesterase (PME) plays an important role in vegetative and reproductive development and biotic/abiotic stress responses by regulating the degree of methyl-esterification of pectic polysaccharides in the plant cell wall. PMEs are encoded by a large multigene family in higher land plant genomes. In general, the expression of plant PME genes shows tissue- or cell-specific patterns and is induced by endogenous and exogenous stimuli. In this study, we identified PME multigene family members (CsPMEs) from the sweet orange genome and report detailed molecular characterization and expression profiling in different citrus tissues and two fruit developmental stages. We also discussed the possible functional roles of some CsPME genes by comparing them with the known functions of PMEs from other plant species. We identified 48 CsPME genes from the citrus genome. A phylogenetic tree analysis revealed that the identified CsPMEs were divided into two groups/types. Some CsPMEs showed very close phylogenetic relationships with the PMEs whose functions were formerly addressed in Arabidopsis, tomato, and maize. Expression profiling showed that some CsPME genes are highly or specifically expressed in the leaf, root, flower, or fruit. Based on the phylogenetic relationships and gene expression profiling results, we suggest that some CsPMEs could play functional roles in pollen development, pollen tube growth, cross incompatibility, root development, embryo/seed development, stomata movement, and biotic/abiotic stress responses. Our results shed light on the biological roles of individual CsPME isoforms and contribute to the search for genetic variations in citrus genetic resources.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.9.1-9.11
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• 2022

Background: Escherichia coli, which causes subclinical or clinical mastitis in cattle, is responsible for transmitting antimicrobial resistance via human consumption of raw milk or raw milk products. Objectives: The objective of this study was to investigate the molecular characteristics of 183 E. coli from bulk tank milk of five different dairy factories in Korea. Methods: The molecular characteristics of E. coli such as serogroup, virulence, antimicrobial resistance, and integron genes were detected using polymerase chain reaction and antimicrobial susceptibility were tested using the disk diffusion test. Results: In the distribution of phylogenetic groups, group D was the most prevalent (59.6%) and followed by group B1 (25.1%). The most predominant serogroup was O173 (15.3%), and a total of 46 different serotypes were detected. The virulence gene found most often was fimH (73.2%), and stx1, fimH, incC, fyuA, and iutA genes were significantly higher in isolates of phylogenetic group B1 compared to phylogenetic groups A, B2, and D (p < 0.05). Among 64 E. coli isolates that showed resistance to at least one antimicrobial, the highest resistance rate was observed for tetracyclines (37.5%). All 18 integron-positive E. coli carried the integron class I (int1) gene, and three different gene cassette arrangements, dfrA12+aadA2 (2 isolates), aac(6')-Ib3+aac(6')-Ib-cr+aadA4 (2 isolates), and dfrA17+aadA5 (1 isolate) were detected. Conclusions: These data suggest that the E. coli from bulk tank milk can be an indicator for dissemination of antimicrobial resistance and virulence factors via cross-contamination.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.210-210
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• 2022

Silybum marianum is an annual or biennial plant from the Asteraceae family. It can grow in low-nutrient soil and drought conditions, making it easy to cultivate. From the seed, a specialized plant metabolite called silymarin (flavonolignan complex) is produced and is known to alleviate the liver from hepatitis and toxins damages. To infer the phylogenetic placement of a Korean milk thistle, we conducted a chloroplast assembly and annotation following by a comparison with existing Chinese reference genome (NC_028027). The chloroplast genome structure was highly similar with an assembly size of 152,642 bp, an 153,202 bp for Korean and Chinese milk thistle respectively. Moreover, there were similarities at the gene level, coding sequence (n = 82), transfer RNA (n = 31) and ribosomal RNA (n = 4). From all coding sequences gene set, the phylogenetic tree inference placed the Korean cultivar into the milk thistle clade; corroborating the expected tree. Moreover, an investigation the tree based only on the ycf1 gene confirmed the same tree; suggesting that ycf1 gene is a potential marker for DNA barcoding and population diversity study in milk thistle genus. Overall, the provided data represents a valuable resource for population genomics and species-centered determination since several species have been reported in the Silybum genus.

• Korean Journal of Plant Taxonomy
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• v.46 no.3
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• pp.314-325
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• 2016

Multiple changes of the photosynthesis pathway are independent evolutionary events occurring in the phylogeny of flowering plants, and such changes have occurred more than five times in Cyperaceae. In the tribe Cypereae, the C4 photosynthetic pathway appeared only once and is regarded as a synapomorphy of the C4 plants within this tribe. The morphological delimitation of genera within Cypereae does not correspond to their molecular phylogenetic relationships. In this study, the molecular phylogeny was compared with the photosynthetic pathways of Korean Cypereae (18 species of Cyperus, 1 species of Kyllinga, and 1 species of Lipocarpha). The photosynthetic pathways were determined by observing the leaf anatomy. The phylogenetic analysis was performed using three DNA regions (nrITS, rbcL, and trnL-F). According to the position of the photosynthetic tissue, 4 species (C. difformis, C. flaccidus, C. haspan, and C. tenuispica) and 16 species (14 Cyperus species, K. brevifolia var. leiolepis, and L. microcephala) were confirmed as C3 and C4 plants, respectively. Tribe Cypereae was divided into the CYPERUS and FICINIA clades, and all species of Korean Cypereae plants belonged to the CYPERUS clade in the phylogenetic analysis. Within the CYPERUS clade, C4 plants were monophyletic but their phylogenetic relationships were unclear. The genera Kyllinga and Lipocarpha were not supported as an independent genus in either case because they were nested by the Cyperus species in the molecular phylogenetic trees in the present and in previous studies. To determine the classification within the CYPERUS clade, a detailed morphological study and a molecular phylogenetic analysis at a high resolution will be necessary.

• Pediatric Infection and Vaccine
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• v.17 no.1
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• pp.16-22
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• 2010

Purpose : We aimed to investigate the clinical and phylogenetic characteristics of Escherichia coli Urinary Tract Infections (E. coli UTI). Methods : We enrolled patients with culture-proven E. coli UTI, who were admitted at the study hospital from September 2008 to August 2009. We investigated clinical data of patients with E. coli UTI and characteristics of isolated E. coli strains. The phylogenetic groups were classified using triplex polymerase chain reaction (PCR), and the distribution of nine virulent genes was determined by multiplex PCR. Results : A total of 47 patients have participated in this study. Thirty (63.8%) were under 6 months; eight (17.0%) were between 6-12 months; and nine (19.1%) were over 12 months. We compared two age groups between under 6-month and over 6-month. In the age group under 6-month, higher proportion of male (P =0.002) and group B2 strains (P =0.020) were observed. In contrast, higher proportion of female and group non-B2 strains were observed in age group over 6-month. Frequencies of papC, papGII, papGIII, sfa/foc, hlyC, cnf1, fyuA, iroN and iucC were estimated as 68.1%, 57.4%, 42.6%, 46.8%, 46.8%, 31.9%, 87.2%, 48.9% and 63.8%, respectively. In the comparison of phylogenetic groups, group B2 showed higher distribution of virulent genes, while group D included more strains resistant to trimethoprim/sulfamethoxazole (TMP/SMZ) than other groups. Conclusion : We showed the age group-specific difference in the distribution of sex ratios and phylogenetic groups; more male and group B2 strains in age group under 6-month, while more female and group non-B2 in age group over 6-month. However, further evaluation including larger number of patients will be necessary to confirm above thesis in future molecular epidemiological studies.

• Journal of Mushroom
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• v.10 no.2
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• pp.93-99
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• 2012

This study was carried to identify a correct species and asses genetic diversity within the same species of Grifola spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Grifola spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Grifola clustered into one group, most of which correlated with species-groups identified by RAPD method. Eight isolates included strain GM01 showed high similarity with Grifola frondosa. All isolates were collected in the Japan(GM01, GM02, GM03) was identified as Grifola frondosa and isolates of the China(GM05, GM06, GM08) was identified as Bjerkandera fumosa, Grifola frondosa and Dichomitus squalens, respectively. RAPD analysis of genetic polymorphisms of genus Grifola showed a very different band patterns on the isolat. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 4 isolates of Grifola spp. may be need to reclassify or eliminate from preserved catalogue.