• Korean Journal of Plant Taxonomy
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• v.36 no.4
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• pp.263-277
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• 2006

Anemone pendulisepala, recently described as a new species, is distributed in sympatry with A. reflexa, A. amurensis, and A. raddeana at Mt. taebeark and Mt. Baekdu. Anemone pendulisepala was previously proposed to be a hybrid species between A. reflexa and A. amurensis becaue it displavs overlapping features with them in involucre shape, petiole length, sepal apex and xylem shape, To verify the taxonomic status and to examine the hybridity of A. pendulisepala, sequences of ITS region of nuclear ribosomal DNA and the psba-trnH, rps16 and trnLF region of cpDNA from 36 accessions of 5 taxa including outgroup were analyzed. In maximum parsimony tree based on ITS sequences, A. pendulisepala had the same sequences of A. reflexa and was clustered with monophyletic A. amurensis, and then A. raddeana. Anemone pendulisepala was distinguished from the other taxa by having four base insertion in rps16 region, two species-specific bases and insertion in trnLF region. In the phylogenetic trees of combined cpDNA, A. pendulisepala showed monophyly with the bootstrap 100%. Anemone pendulisepala exhibited no polymorphism and shared no sequences with putative parental or related taxa examined in this study. Molecular data suggest that A. pendulisepala should be a distinct species, and no evidence of the hybridization was detectcd.

• Journal of Animal Science and Technology
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• v.53 no.4
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• pp.303-310
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• 2011

Phylogenetic relationships of Jeju dogs to other domestic and foreign dog breeds were assessed using mtDNA D-loop sequences. Neighbor-joining trees were constructed using complete sequences (970 bp excluding the tandem repeat region) determined for five Cheju, four Jindo, four Sapsaree, five Pungsan, two of each East and West Laika dogs (Canis familiaris), two gray wolves (Canis lupus) and two coyotes (Canis latrans) and also published complete sequences for dogs. Coyote sequences were used as outgroups. In addition, a total of 214 haplotypes of 598bp D-loop sequences from 30 dog breeds were collected from GenBank and used to investigate genetic structure of population. In the analyses of full D-loop sequence variation and the phylogenetic trees constructed by neighbor-joining method, neither haplotypes nor clades specific for any domestic dog breeds were observed. The inter-species sequence variation (4.51%) between domestic dogs and wolves was much higher than the intra-species sequence variation within domestic dogs (1.63%) and wolves (3.64%). The divergence of the dog and wolf occurred approximately 1~2 million years ago based on these values. The taxa of Jeju dog breed in the phylogenetic tree are clustered separately and intermingled with other taxa of breeds, suggesting that active crossbreeding of Jeju dogs with other domestic breeds.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.2
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• pp.143-149
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• 2000

From the soils of soybean fields in Cotton Branch Station (CBS) and Pine Tree Station (PTS), Arkansas, USA, various single spore isloates of sudden death syndrome (SDS) pathogen were obtained on modified Nash ＆ Snyder's medium (MNSM) with dilution plating technique and transferred to potato dextrose agar (PDA) medium to identify the cultural colony shape. The colony shapes of these isolates resembled F. solani isolate 171 which was white and chalky shaped on MNSM and most of them had unique form of morphology which produced white margin and blue center colony on PDA. Although, some of these isolates had more dark blue or showed slightly different color, all isolates that were selected randomly for green-house inoculation assay produced typical foliar symptoms on leaves of soybean, Hartz 6686. To determine the genetic differences among the isolates, mitochondrial DNA restriction fragment length polymorphism (RFLP) was conducted with fourty isolates from both fields, using mtDNA probes, 2U18 and 4U40, derived from Colletotrichum orbiculare. We obtained distinctive RFLPs in each treatment of restriction enzyme, EcoRI and HaeⅢ. Isolates, 11-2-5 and 14-3-1-1, from CBS and isolates, 104-3-1-2 and 701-1-5-1, from PTS showed different band patterns from 171 in both or in either treatment of restriction enzymes. Even if some of these isolates showed heterogeneous, they were more closer to 171 than PN603. And, also, rest of the thirty-six isolates had exactly same polymorphisms as 171 in each treatment of restriction enzyme. Although, some of the isolates showed the different morphological shape on PDA and slightly different band patterns on RFLPs, all of the isolates selected on MNSM due to their distinctive colony shape from other fungi produced the typical foliar symptoms on soybean leaves in greenhouse inoculation assay. It might be suggested that these isolates were not genetically different from check isolate 171 and they were unique strain of F. solani.

• Parasites, Hosts and Diseases
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• v.47 no.4
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• pp.337-344
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• 2009

In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the $\alpha$-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.41-41
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• 2015

Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

• Journal of Life Science
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• v.24 no.6
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• pp.626-632
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• 2014

The genetic structure and phylogenetic relationship were investigated in Korean red spotted grouper populations using the nucleotide sequence polymorphisms of the mitochondrial DNA (mtDNA) cytochrome c oxidase subunit I (COI) gene. The COI gene was sequenced showed 99.1-99.8% identity with the EF607565 sequence previously reported. A total of twenty haplotypes were found, and the Korean population showed nineteen haplotypes. Among those, Hap_03 and Hap_08 showed Jeju-do and China-specific COI sequences, respectively. However, Hap_07 had twelve COI sequences from South Korea and records from Hong Kong and Taiwan. Neighbor-joining (NJ) trees constructed from the phylogenetic analyses based on the polymorphisms of the COI haplotypes showed a monophyletic branching pattern within the genus Epinephelus. This indicated that the red spotted grouper populations had evolved from common maternal ancestors. In addition, the Hap_08, which had the COI sequence recorded only from China Sea, was found in the middle of the NJ tree nearby Hap_07 and showed a close relationship with Hap_07. This indicates that Chinese red spotted grouper is also maternally related to other populations in East Asia. Consequently, East Asian red spotted grouper populations are maternally related, as well as sharing the same evolutionary history, and are still affected by the East Asian ocean current (Kuroshio). These findings help to explain the genetic structure and phylogenetic relationship of red spotted grouper and also contribute to research on artificial breeding and industrialization.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.6
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• pp.967-972
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• 2022

We investigated the molecular and morphological traits of 338 individual macrourids collected from the Southern Ocean (FAO area number, 88.1 and 88.3) between 2021 and 2022 by Korean bottom trawls. We first identified them as Macrourus caml and Macrourus whitsoni based on morphological traits, such as the number of pelvic fin rays (PF) and the rows of lower jaw teeth (LJT). However four individuals showed uncategorizable morphological characteristics such as PF and LJT numbers that overlapped between the two species. Subsequently, we obtained and analyzed 509 bp of the mtDNA COI sequences of 49 individuals, including the four unidentified individuals, and found only one single nucleotide polymorphism (SNP) that distinguished the two species. Finally, using our molecular identification key, we confirmed that each two individuals were misidentified as M. whitsoni and M. caml reversely. Our results suggest that the number of PF and LJT should be investigated together to accurately identify the two species.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.8
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• pp.1066-1074
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• 2015

In contrast to high genetic diversity of mitochondrial DNA (mtDNA), equine Y chromosome shows extremely low variability, implying limited patrilines in the domesticated horse. In this study, we applied direct sequencing and restriction fragment length polymorphism (RFLP) methods to investigate the polymorphisms of 33 Y chromosome specific loci in 304 Chinese indigenous horses from 13 breeds. Consequently, two Y-single nucleotide polymorphisms (SNPs) (Y-45701/997 and Y-50869) and one Y-indel (Y-45288) were identified. Of those, the Y-50869 (T>A) revealed the highest variation frequency (24.67%), whereas it was only 3.29% and 1.97% in Y-45288 (T/-) and Y-45701/997 (G>T) locus, respectively. These three mutations accounted for 27.96% of the total samples and identified five Y-SNP haplotypes, demonstrating genetic diversity of Y chromosome in Chinese horses. In addition, all the five YSNP haplotypes were shared by different breeds. Among 13 horse breeds analyzed, Balikun horse displayed the highest nucleotide diversity (${\pi}=5.6{\times}10^{-4}$) and haplotype diversity (h = 0.527), while Ningqiang horse showed the lowest nucleotide diversity (${\pi}=0.00000$) and haplotype diversity (h = 0.000). The results also revealed that Chinese horses had a different polymorphic pattern of Y chromosome from European and American horses. In conclusion, Chinese horses revealed genetic diversity of Y chromosome, however more efforts should be made to better understand the domestication and paternal origin of Chinese indigenous horses.

• Journal of Life Science
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• v.16 no.5
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• pp.812-827
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• 2006

To investigate the population structure and geographic distance among anchovies (Engraulis japonicus) in Korea, we compared and analyzed the mitochondrial DNA control region sequences (227 bp) of anchovies from 12 localities in inshore and offshore waters. The sequence analysis of 84 individuals showed 29 haplotypes, ranging in sequence divergence by pairwise comparisons from 0.3% to 3.5% (1 bp-12 bp). E9 haplotype of anchovies were found largely in inshore waters and also in offshore waters, which was regarded as the major source in Korean waters (58.3%). However, E26, E27, E28, and E29 haplotypes were found in westsouthern (locality 10, four among 7). Phylogenetic analysis using PHYLIP was divided into two clades (clade A and B). Most of the haplotypes, excluding E26, E27, E28, and E29, were strongly supported by bootstrap analysis (>75%), whereas the relationship between clade A and B was weakly supported by bootstrap analysis (51%). High levels of genetic diversity were found; haplotype diversity (H)=0.75-1.00, and nucleotide diversity $({\pi})=0.015-0.0244$. Analysis of $F_{ST}$ between populations in inshore waters ranged in 0.01-0.05, whereas those of offshore waters ranged in 0.01-0.58. A high gene flow occurred in inshore (Nm=22.61-34.22) and offshore (Nm=11.57-45.67) populations. The distribution of mitochondrial DNA haplotypes between westsouthern and other populations was suggestive of significantly different differentiation ($F_{ST}$=0.20-0.59, p<0.05; d=0.52, p=0.00; ${\phi}=0.02-0.41$, p＜0.05). These results suggested that the overall anchovy population in the Korean peninsula caused considerable migration due to the mitochondrial gene flow between inshore and offshore populations to form a genetically homogenous and panmictic structure, although a heterogeneous population was found in this study.

• Korean Journal of Ichthyology
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• v.33 no.2
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• pp.45-56
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• 2021

A male, presumed to be an intergeneric hybrid between Tanakia lanceolata and Rhodeus pseudosericeus, was collected in the Boryeong Daecheoncheon Stream flowing into the Yellow Sea in the Republic of Korea. Morphological and molecular phylogenetic analyses were performed to discriminate the definite origin of the estimated natural hybrid. As a result of the morphological analysis, the color of the dorsal and anal fin rays edges of the natural hybrid individual, the upper and lower body colors followed the morphological characteristics of T. lanceolata, and that blue longitudinal stripe in the center of the caudal peduncle, the incomplete lateral line, and the barbels absent followed the morphological characteristics of R. pseudosericeus. In addition, as a result of the cytochrome b (cytb) gene analysis of mitochondrial DNA (mtDNA), the natural hybrid showed a nucleotide sequence similarity of 99.82 to 100% with T. lanceolata, and the maternal species was identified as T. lanceolata. As a result of the recombination activating gene 1 (rag1) gene analysis of nuclear DNA (nDNA), the natural hybrid showed double peaks pattern reflecting both the single nucleotide polymorphism sites (38 bp) between T. lanceolata and R. pseudosericeus, and the paternal species was identified as R. pseudosericeus. Therefore, a natural hybrid estimated male of Acheilognathinae analyzed in this study was found to be an intergeneric hybrid between a female T. lanceolata and a male R. pseudosericeus.