• The Korea Journal of Herbology
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• v.34 no.5
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• pp.13-20
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• 2019

Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

• Korean Journal of Ecology and Environment
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• v.55 no.4
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• pp.352-359
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• 2022

Recently, with the development of genetic technology, interest in environmental DNA (eDNA) to study biodiversity according to molecular biological approaches is increasing. Environmental DNA has many advantages over traditional research methods for biological communities distributed in the environment but highly depends on the established base sequence database. This study conducted a comprehensive analysis of the habitat status and classification at the genus level, which is mainly used in eDNA (12S rRNA, 16S rRNA, 18S rRNA, COI, and CYTB), focusing on Korean registration taxon groups (phytoplankton, zooplankton, macroinvertebrates, and fish). As a result, phytoplankton and zooplankton showed the highest taxa proportion in 18S rRNA, and macroinvertebrates observed the highest ratio in the nucleotide sequence database in COI. In fish, all genes except 18S rRNA showed a high taxon ratio. Based on the Korean registration taxon group, the gene construction of the top 20 genera according to bio density observed that most of the phytoplankton were registered in 18S rRNA, and the most significant number of COI nucleotide sequences were established in macroinvertebrates. In addition, it was confirmed that there is a nucleotide sequence for the top 20 genera in 12S rRNA, 16S rRNA, and CYTB in fish. These results provided comprehensive information on the genes suitable for eDNA research for each taxon group.

• Korean journal of applied entomology
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• v.62 no.4
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• pp.227-243
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• 2023

The Asiatic pink stem borer, Sesamia inferens (Walker, 1856) (Lepidoptera: Noctuidae) is a pest that attacks rice stems. In this study, we estimated the annual generation of insect in several regions of the Republic of Korea. Adult trapping using a sex pheromone trap detected the occurrence of S. inferens adults in the three northernmost areas around 38° latitude and showed that the insect inhabits all of Korea. In most areas investigated, the seasons of the adult generations estimated using the single-sine degree-day model did not deviate from the corresponding observed seasons of adult occurrence. We estimated that the overwintering larvae hypothetically-estimated using the model could be originated from the last generation of adults. When larvae collected in paddy fields during the autumn season in a few middle and southern areas were reared at 25℃, ca. 70% of pupae did not show additional larval molting before their pupation. All larvae collected in early March in a southern area (Goseong, Gyeongsangnam-do) pupated without additional larval molting when reared at 25℃. Based on these results, we presumed that S. inferens could overwinter as mainly the last instar larval stage in the area. Taken together, we conclude that S. inferens primarily has two generations per year in areas around 38° latitude, and three generations in the areas between 35.3° and 37.3° latitude. In addition, approximately 35% of insects captured by the sex pheromone trap were species other than S. inferens, as determined by analyzing the nucleotide sequences of the cytochrome c oxidase 1 gene. These species were not morphologically misidentified as S. inferens.

• Korean Journal of Environment and Ecology
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• v.29 no.5
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• pp.693-702
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• 2015

In order to identify the species and to reveal the phylogenetic relationship of rook populations found in Jeju-do Province in winter seasons, we determined the sequences of mitochondrial cytochrome c oxidase I (COI) gene and analyzed the genetic structure of maternal lineages and phylogenetic relationship. The rook DNAs were isolated from the post-mortem specimens and plumages collected from agricultural farms in Jeju-do Province including U-do Island. The obtained COI sequences (n=41) showed over 97.0% identities with those previously reported from Corvus frugeligus. Three COI haplotypes (J01-J03) were detected from COI sequences of the rooks obtained in Jeju-do Province but those did not show the site-specific patterns, showing that they might be derived from a common maternal origin. Eight maternal haplotypes were detected from all COI sequences obtained. Among those three haplotypes contained the COI sequences from Northeast Asia including eastern Russia, Mongolia and South Korea. On the other hand, the other five haplotypes contained the COI sequences reported from Central Asia, Middle East, western Russia and European countries. The COI sequences from Jeju-do Province were located on three haplotypes (CF01-CF03) belonging to Northeast Asian rook lineages. The NJ tree showed the distinct branch patterns suggesting two different maternal lineages of C. frugilegus, which proposed as two parapatric subspecies, C. f. frugilegus (Western) and C. f. pastinator (Eastern). These findings using DNA barcoding approaches will be contributed to provide the information about avian fauna for understanding the genetic structure of maternal lineage, phylogenetic relationship and their molecular ecology.

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.347-355
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• 2014

In this study, the detection method was developed using molecular biological technique to distinguish authenticity of animal raw materials. The genes for distinction of species about animals targeted at Cytochrome c oxidase subunit I (COI), Cytochrome b (Cytb), and 16S ribosomal RNA (16S rRNA) genes in mitochondrial DNA. The species-specific primers were designed by that Polymerase Chain Reaction (PCR) product size was around 200 bp for applying to processed products. The target 24 raw materials were 2 species of domestic animals, 6 species of poultry, 2 species of freshwater fishes, 13 species of marine fishes and 1 species of crustaceans. The results of PCR for Rabbit, Fox, Pheasant, Domestic Pigeon, Rufous Turtle Dove, Quail, Tree Sparrow, Barn Swallow, Catfish, Mandarin Fish, Flying Fish, Mallotus villosus, Pacific Herring, Sand Lance, Japanese Anchovy, Small Yellow Croaker, Halibut, Jacopever, Skate Ray, Ray, File Fish, Sea Bass, Sea Urchin, and Lobster raw materials were confirmed 113 bp ~ 218 bp, respectively. Also, non-specific PCR products were not detected in compare species by species-specific primers. The method using primers developed in this study may be applied to distinguish an authenticity of food materials included animal raw materials for various processed products.

• 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.

• Journal of Nutrition and Health
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• v.57 no.1
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• pp.53-64
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• 2024

Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.

• Korean journal of applied entomology
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• v.59 no.2
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• pp.93-107
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• 2020

Adults of seven noctuid potential pests (Spodoptera frugiperda, S. litura, S. exigua, Ctenoplusia agnata, Mythimna loreyi, Athetis dissimilis, and A. lepigone) of soybean and maize in Suwon, Korea were identified by their morphological characteristics in the wing pattern and male genitalia and partial mitochondrial DNA sequences of cytochrome c oxidase subunit 1 gene. The generation number of adults that emerge annually in six species (except A. lepigone) was estimated from the data on density fluctuations of adults caught in sex pheromone traps in 2019 and the forecasted data using temperature-associated development and reproduction models for those species. S. frugiperda adults were caught from July 27th to October 31st in 2019, and hence were initially estimated to emerge three times per year. But, it was finally expected that S. frugiperda adults could possibly emerge a total of four times per year in Suwon, considering larval emergence observed during mid- and late June in other areas. Adult emergence of S. litura, S. exigua, C. agnata, and M. loreyi in 2019 was observed from May 29th to November 6th, from May 14th to November 6th, from May 26th to October 25th, and from May 31st to November 23rd, respectively. Annual adult emergence of these four species was estimated as at least four times. Adults of A. dissimilis were caught from May 26th to September 11th in 2019, and adult emergence was estimated at only twice per annum. It was postulated that the first adult populations of five species except the two Athetis species were probably migrated from other areas.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.263-268
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• 2016

Metallothionein (MT) family of metal-binding proteins are involved in maintaining homeostasis and heavy metal poisoning. Recently, MT has been considered as a biomarker that can identify a particular species, very similar to the use of cytochrome oxidase I (COI) gene. Satsuma myomphala species of land snails have been reported from North-East Asia, including South Korea and Japan. In particular, the land snail species have been known from only a limited area of Geoje Island, Gyeongsangnam-do province of South Korea. Genetic studies of S. myomphala has been limited with only 6 nucleotide, 2 protein registered on the NCBI server. For elucidating the genetic information of S. myomphala, we conducted RNA sequencing analysis using Illumina HiSeq 2500 next-generation platform. We screened the MT gene from the RNA-Seq database to confirm the molecular phylogenetic relationship. After sequencing, the de novo analysis and clustering generated 103,774 unigenes. After annotation against PANM database using BLAST program, we obtained MT sequence of 74 amino acid residues containing the coding region of 222 bp. Based on this sequence, we found about 53 sequences using the BLAST program in NCBI nr database. Using ClustalX alignment, Maximum-Likehood Tree of MEGA program, we confirmed the molecular phylogenetic relationships that showed similarity with mollusks such as Helix pomatia and H. aspersa, Megathura crenulata.