• Animal Systematics, Evolution and Diversity
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• v.37 no.1
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• pp.26-30
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• 2021

Botrylloides violaceus is native to the Northwest Pacific, including Korea. This species has many color variations in alive condition and a variety of zooid compound forms, and therefore difficult to identification in the field survey. This is the first report of COI DNA barcodes of B. violaceus from Korea. The intra-specific pairwise distance between Korean and UK populations had ranged from 1.4% to 2.6%. The inter-specific variations between B. violaceus and other Botrylloides species were 21.0-36.8%. The new DNA barcodes for Korean B. violaceus may be helpful in the identification of colonial ascidians, which is a difficult task when based on morphological identification.

• 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 Plant Biotechnology
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• v.48 no.3
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• pp.131-138
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• 2021

We reviewed current research trends for discriminating between species of the Angelica genus, a group of important medicinal plants registered in South Korea, China, and Japan. Since the registered species for medicinal purposes differ by country, they are often adulterated as well as mixed in commercial markets. Several DNA technologies have been applied to distinguish between species. However, one of the restrictions is insufficient single-nucleotide polymorphisms (SNPs) within the target DNA fragments; in particular, among closely-related species. Recently, amplification refractory mutation system (ARMS)-PCR and highresolution melting (HRM) curve analysis techniques have been developed to solve such a problem. We applied both technologies, and found they were able to discriminate several lines of Angelica genus, including A. gigas Nakai, A. gigas Jiri, A. sinensis, A. acutiloba Kitag, and Levisticum officinale. Furthermore, although the ITS region differs only by one SNP between A. gigas Nakai and A. gigas Jiri, both HRM and ARMS-PCR techniques were powerful enough to discriminate between them. Since both A. gigas Nakai and A. gigas Jiri are native species to South Korea and are very closely related, they are difficult to discriminate by their morphological characteristics. For practical applications of these technologies, further research is necessary with various materials, such as dried or processed materials (jam, jelly, juice, medicinal decoctions, etc.) in commercial markets.

• The Korea Journal of Herbology
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• v.34 no.4
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• pp.1-8
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• 2019

Objectives : Rosae Multiflorae Fructus is a traditional medicine derived from the fruit of Rosa multiflora Thunb. a member of the Rosaceae family. Even though it has a single origin, the possibility of adulterants has always existed. In fact, we had discovered suspicious commercial samples of Rosae Multiflorae Fructus, imported from China. Methods : To define the taxonomic origin of Rosae Multiflorae Fructus and its adulterants, DNA barcode analysis of the internal transcribed spacer, trnL-F intergenic spacer, and psbA-trnH sequences was carried out. These DNA barcode sequences from the correct origin of Rosae Multiflorae Fructus were analyzed and compared with those of other samples from genus Rosa used as medicinal herbs. Results : The analyses of the three DNA barcode sequences efficiently distinguished Rosae Multiflorae Fructus from six other species in genus Rosa and also separated each species used in this study. According to the DNA barcoding results, none of the suspicious commercial samples were Rosae Multiflorae Fructus. RMF09 was identified as Rosa acicularis, whereas RMF10 and RMF11 were identified as Rosa davurica and Rosa rugosa, respectively. These results corroborated the existence of adulterants of Rosae Multiflorae Fructus. Conclusions : Our research provides useful information that could be used as a criterion for distinguishing between Rosae Multiflorae Fructus and its adulterants. These results will help in the prevention of adulteration and also suggest effective methods for verifying the origin of commercial herbal medicines derived from genus Rosa.

• 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.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.17-17
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• 2014

Over five thousand higher fungal specimens were collected from 32 forest areas of Chungcheong and Gyeongsang province from 2012 to 2013. We obtained 513 strains and 3,120 ITS sequences. Mushrooms were first identified with macro- and micro-scopic characters, and their identification was confirmed on the basis of ITS sequences. Voucher specimens were designated for each species found in Korea. Construction of DNA barcoding Database is currently underway with sequences of 409 species. During the development of the database, some new species were recognized, along with several Korean new records. When the system has been completed, it will provide essential molecular information for metagenomic and phylogenetic researches for higher fungi.

• International Journal of Industrial Entomology and Biomaterials
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• v.31 no.2
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• pp.40-47
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• 2015

The genus Acteniceromorphus is taxonomically reviewed for the first time in Korea. From the previously recorded three species, two species, A. selectus (Candèze, 1894) and A. fulvipennis (Lewis, 1894) are confirmed as misidientification of species which are endemic to Japan. Another species, A. chlamydatus (Lewis, 1894) is unavailable any Korean specimens. Additionally, we found A. ferrugineipennis (Candèze, 1879) as new to Korea. We also provide a comparison of DNA barcoding for two species previously misidentified and the newly recorded species, except A. chlamydatus. DNA barcoding result shows that each species is clearly delimited at species-level from each other by large interspecific genetic distance over 7.2%.

• Animal Systematics, Evolution and Diversity
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• v.35 no.3
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• pp.156-159
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• 2019

The phyllodicd polychaete species, Nereiphylla hera Kato and Mawatari, 1999 is reported from the intertidal habitats of the eastern coast of South Korea. We determined the DNA barcoding region of the mitochondrial cytochrome c oxidase subunit I (COI) of N. hera and compared nucleotide variation with its congeners. The intra-specific genetic distance between the three COI sequences of N. hera was ranged from 0 to 0.4%. The inter-specific distances between N. hera and other Nereiphylla species ranged from 18.8 to 22.3%. In this study, we reported the first COI barcodes of N. hera with the morphologcial diagnosis and the photographs. These results would be helpful to understand taxonomy of Nereiphylla.

• Animal Systematics, Evolution and Diversity
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• v.38 no.2
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• pp.108-112
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• 2022

An aegid species, Rocinela niponia Richardson, 1909, is a Far Eastern species known from Korean and Japanese waters. In this study, mitochondrial cytochrome c oxidase subunit I (COI) sequences of R. niponia were determined based on four specimens collected from the subtidal zone of Chujado Island, South Korea. We compared DNA barcoding data of this species with its congeners. As a result, there was no intra-specific genetic distance between the four COI sequences of R. niponia. Inter-specific distances between R. niponia and other five aegid species ranged from 23.8% to 35.6%. Morphological diagnosis and images of R. niponia are also provided as a valuable contribution toward the identification of Rocinela species in further taxonomic and ecological studies.

• Korean Journal of Ecology and Environment
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• v.54 no.3
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• pp.156-169
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• 2021

Traditional morphological identification difficulties, such as phenotypic plasticity, misidentification of cryptic species, and larval stage species, can be compensated for by using DNA analysis techniques, such as DNA barcoding, in surveying zooplankton populations, including species identification. Recently, the rapid development of DNA sequencing techniques has allowed DNA-based community analysis not only for zooplankton assemblages in various aquatic ecosystems but also for the gut contents of zooplankton that are limited by conventional methods such as visual and microscopic identification. Therefore, the application of DNA sequencing can help understand biological interactions through the analysis of zooplankton food sources. The present paper introduces the major DNA-based approaches in zooplankton research topics, including taxonomic approaches by DNA barcoding, community-level approaches by metabarcoding, and gut content analyses, summarizes the analysis methods, and finally suggests the methodological topics that need to be considered for future applications.