• Animal Systematics, Evolution and Diversity
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• v.28 no.4
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• pp.269-278
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• 2012

Gobiids are hyperdiverse compared with other teleost groups, with about 2,000 species occurring in marine, freshwater, and blackish habitats, and they show a remarkable variety of morphologies and ecology. Testing the effectiveness of DNA barcodes on species that have emerged as a result of radiation remains a major challenge in evolutionary biology. Here, we used the cytochrome c oxidase subunit 1 (COI) sequences from 144 species of gobies and related species to evaluate the performance of distance-based DNA barcoding and to conduct a phylogenetic analysis. The average intra-genus genetic distance was considerably higher than that obtained in previous studies. Additionally, the interspecific divergence at higher taxonomic levels was not significantly different from that at the intragenus level, suggesting that congeneric gobies possess substantial interspecific sequence divergence in their COI gene. However, levels of intragenus divergence varied greatly among genera, and we do not provide sufficient evidence for using COI for cryptic species delimitation. Significantly more nucleotide changes were observed at the third codon position than that at the first and the second codons, revealing that extensive variation in COI reflects synonymous changes and little protein level variation. Despite clear signatures in several genera, the COI sequences did resolve genealogical relationships in the phylogenetic analysis well. Our results support the validity of COI barcoding for gobiid species identification, but the utilization of more gene regions will assist to offer a more robust gobiid species phylogeny.

• Korean Journal of Medicinal Crop Science
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• v.23 no.6
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• pp.439-445
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• 2015

Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

• ALGAE
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• v.35 no.3
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• pp.293-301
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• 2020

The applications of DNA barcoding have a wide range of uses, such as in taxonomic studies to help elucidate cryptic species and phylogenetic relationships and analyzing environmental samples for biodiversity monitoring and conservation assessments of species. After obtaining the DNA barcode sequences, sequence similarity-based homology analysis is commonly used. This means that the obtained barcode sequences are compared to the DNA barcode reference databases. This bioinformatic analysis necessarily implies that the overall quantity and quality of the reference databases must be stringently monitored to not have an adverse impact on the accuracy of species identification. With the development of next-generation sequencing techniques, a noticeably large number of DNA barcode sequences have been produced and are stored in online databases, but their degree of validity, accuracy, and reliability have not been extensively investigated. In this study, we investigated the extent to which the amount and types of erroneous barcode sequences were deposited in publicly accessible databases. Over 4.1 million sequences were investigated in three largescale DNA barcode databases (NCBI GenBank, Barcode of Life Data System [BOLD], and Protist Ribosomal Reference database [PR2]) for four major DNA barcodes (cytochrome c oxidase subunit 1 [COI], internal transcribed spacer [ITS], ribulose bisphosphate carboxylase large chain [rbcL], and 18S ribosomal RNA [18S rRNA]); approximately 2% of erroneous barcode sequences were found and their taxonomic distributions were uneven. Consequently, our present findings provide compelling evidence of data quality problems along with insufficient and unreliable annotation of taxonomic data in DNA barcode databases. Therefore, we suggest that if ambiguous taxa are presented during barcoding analysis, further validation with other DNA barcode loci or morphological characters should be mandated.

• International Journal of Industrial Entomology and Biomaterials
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• v.35 no.1
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• pp.51-57
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• 2017

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae), is an endangered insect in South Korea. Previously, a partial mitochondrial DNA sequence that corresponded to a DNA barcoding region has been used to infer genetic diversity and gene flow. In this study, we additionally sequenced the barcoding region from N. pygmaea that had been collected from three previously sampled populations (40 individuals) and these sequences were combined with the preexisting data. We also selected and sequenced an additional mitochondrial gene (ND5) to find further variable gene regions in the mitochondrial genome. DNA barcoding sequences of 108 individuals from five South Korean localities showed that genetic diversity was highest in Gangjin, Jeollanam-do Province. Muuido, which was previously occupied by a single haplotype, was also found to have an identical haplotype, which confirmed the low genetic diversity on this islet. Gene flow among populations is highly limited, and no clear distance- or region-based geographic partitioning was observed. Phylogenetic relationships among haplotypes showed that there were no discernable haplotypes in South Korea. ND5 provided slightly more haplotypes compared to the barcoding region in 40 individuals (14 vs. 10 haplotypes in the COI gene). It also had a slightly higher within-locality diversity estimate, which suggested that ND5 had potential as mitochondrial DNA-based marker for population genetic analysis.

• Fisheries and Aquatic Sciences
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• v.27 no.2
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• pp.87-99
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• 2024

Global climate change, followed by an increase in anthropogenic activities in aquatic ecosystems, and species invasions, has resulted in a decline in aquatic organism biodiversity. The Batanghari River, Sumatra's longest river, is polluted by mercury-containing illegal gold mining waste (PETI), industrial pollution, and domestic waste. Several studies have provided evidence suggesting a decline in fish biodiversity within the Batanghari River. However, a comprehensive evaluation of the present status of biodiversity in this river is currently lacking. The species under investigation were identified through various molecular-based identification methods, as well as morphological identification, which involved the use of neighbor-joining (NJ) trees. All collected specimens were initially identified using morphological techniques and subsequently confirmed with molecular barcoding analysis. Morphological and DNA barcoding identification categorized all specimens (1,692) into 36 species, 30 genera and 16 families, representing five orders. A total of 36 DNA barcodes were generated from 30 genera using a 650-bp-long fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene. Based on the Kimura two-parameter model (K2P), The minimum and maximum genetic divergences based on K2P distance were 0.003 and 0.331, respectively, and the average genetic divergence within genera, families, and orders was 0.05, 0.12, 0.16 respectively. In addition, the average interspecific distance was approximately 2.17 times higher than the mean intraspecific distance. Our results showed that the COI barcode enabled accurate fish species identification in the Batanghari River. Furthermore, the present work will establish a comprehensive DNA barcode library for freshwater fishes along Batanghari River and be significantly useful in future efforts to monitor, conserve, and manage fisheries in Indonesia.

• 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.

• 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.

• The Korea Journal of Herbology
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• v.37 no.5
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• pp.9-16
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• 2022

Objectives : The purpose of this study is to report that applying the genetic discrimination method to Pinelliae Tuber is suitable as a countermeasure for the limitations of morphological identification announced publicly in the Ministry of Food and Drug Safety(MFDS). Methods : Randomly selected fifty samples in Pinelliae Tuber imported from China were used for morphological and genetic identification. The morphological identification was applied method announced publicly by the MFDS. The traits of morphological identification were classified as Pinellia ternata, P. tripartita, Pinellia pedatisecta, and Typhonium flagelliforme, according to the formation of tuberous root and tuber morphology. The genetic identifications were conducted by Sequence Characterized Amplified Region(SCAR) marker and DNA barcoding analysis for cross-validation, respectively. SCAR marker was verified according to the presence or absence of amplicon through PCR amplification using species-specific primers. DNA barcoding analysis used sequence information of the matK region. Results : As a result of the morphological identification, 27 out of 50 samples were identified as original species 'P. ternata' of genuine 'Pinelliae Tuber', and 23 were identified as adulterant species 'P. pedatisecta'. Unlike this, the genetic identification was identified as the original species 'P. ternata' in all 50 samples in the SCAR marker and matK regional sequence analysis. Conclusions : Pinelliae Tuber of morphological mutant that can not be classified by morphological identification is imported from China. The SCAR marker would be used as accurate and efficient assays for species identification of the morphological mutant.

• 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.

• Mycobiology
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• v.48 no.5
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• pp.331-340
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• 2020

The family Peronosporaceae, an obligate biotrophic group of Oomycota, causes downy mildew disease on many cultivated and ornamental plants such as beet, cucumber, grape, onion, rose, spinach, and sunflower. To investigate the diversity of Peronosporaceae species in Korea, we performed morphological analysis for dried plant herbariums with downy mildew infections by two largest genera, Peronospora and Plasmopara. As a result, it was confirmed that there are five species of Peronospora and two species of Plasmopara, which have been so far unrecorded in Korea, as well as rarely known in the world; Pl. angustiterminalis (ex Xanthium strumarium), Pl. siegesbeckiae (ex Siegesbeckia glabrescens), P. chenopodii-ambrosioidis (ex Chenopodium ambrosioides), P. chenopodii-ficifolii (ex Chenopodium ficifolium), P. clinopodii (ex Clinopodium cf. vulgare), P. elsholtziae (ex Elsholtzia ciliata), and P. lathyrina (ex Lathyrus japonicus). In addition, their phylogenetic relationship was inferred by molecular sequence analysis of ITS, LSU rDNA, and cox2 mtDNA. By rediscovering the seven missing species and barcoding their DNA sequences, this study provides valuable insights into the diversity and evolutionary studies of downy mildew pathogens.