• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.98-98
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• 2019

제니(薺苨, Adenophorae Remotiflori Radix)는 "대한민국약전외한약(생약)규격집(KHP)"에 모시대(Adenophora remotiflorus Miquel)의 뿌리로 수재되어있으나, 형태학적으로 유사한 잔대(A. triphylla), 당잔대(A. stricta) 및 더덕(Codonopsis lanceolata)과 오 혼용 우려가 있어 이들을 구별하기 위한 정확하고 객관적인 종 감별법이 필요하다. 본 연구에서는 '제니'의 기원인 모시대와 오 혼용 우려가 있는 종들을 구별 할 수 있는 유전자 마커를 개발하기 위하여 Genbank에 등록된 ycf2 구간을 활요하여 모시대와 잔대, 당잔대를 구분 할 수 있는 INDEL (insertion/deletion) 마커를 개발하였다. 또한, 보다 정확한 종감별을 위해 DNA 바코드로 활용되고 있는 유전자 부위의 염기서열을 분석하여 ITS (25%), atpB-rbcL (15%), atpF-atpH (14%), rpl16 (13%), trnL-F (10%), matK (9%), rpoC1 (7%)에서 변이율(percent of variable sites)을 확인하였다. 향후, 본 연구에서 개발된 INDEL 마커와 더불어 추가적으로 개발을 진행 중인 분자 마커는 한약재 '제니'의 품질관리에 활용 가능할 것으로 사료된다.

• The Korea Journal of Herbology
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• v.39 no.2
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• pp.1-9
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• 2024

Objectives : Paeng-hwal is described as an insect herbal medicine used for digestive diseases in the Dong-ui-bo-gam. The origin of this herbal medicine is limited to several small crabs, such as Helice tridens. These crab species cohabitat in the same environment and share similar morphological characteristics, making it very difficult to distinguish and collect the individual species for use in dietary supplements or herbal medicines. This study was conducted to develop a genetic identification tool for discriminating among these closely related small crab species. Methods : CO1 DNA barcode regions of 15 samples from 6 species of small crabs were analyzed to obtain the individual sequences. To identify the correct species, comparative analyses were carried out using the database of the NCBI GenBank and the NIBR. SCAR primers were designed to develop simple and rapid assay methods using inter-species specific sequences. Optimal SCAR assay conditions were established through gradient PCR, and the limit of detection (LOD) was determined. Results : Six species of small crabs (Helicana tridens, Macrophthalmus abbreviatus, Helicana tientsinensis, Helicana wuana, Chiromantes dehaani, and Hemigrapsus penicillatus), which are distributed as Paeng-hwal, were identified through CO1 sequences analysis. We also developed SCAR markers to distinguish between six small crabs at the species level. Furthermore, we established the optimal PCR assay methods and the LOD of each individual species. Conclusions : The rapid and simple SCAR-PCR assay methods were developed to identify the species and control the quality of herbal medicines for Paeng-hwal based on the genetic analyses of CO1 DNA barcodes.

• The Korea Journal of Herbology
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• v.30 no.3
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• pp.41-47
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• 2015

Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.

• 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.29 no.6
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• pp.45-53
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• 2014

Objectives : Due to the morphological similarity of in the roots of herbal medicine, the official herbal medicine is very difficult to authenticate between the original plants of Patriniae Radix and two adulterant Patrinia species. Therefore, we introduced DNA barcode analysis to establish a powerful tool for the authentication of Patriniae Radix from its adulterants. Methods : To analyze DNA barcode regions, genomic DNA was extracted from twenty-nine specimens of Patrinia scabiosaefolia, Patrinia villosa, Patrinia saniculifolia, and Patrinia rupestris, and internal transcribed spacer 2(ITS2), matK and rbcL genes were amplified. For identification of species specific sequences, a comparative analysis was performed by the ClastalW based on entire sequences of ITS2, matK and rbcL genes, respectively. Results : In comparison of three DNA barcode sequences, we identified 22, 22, and 12 species-specific nucleotides enough to distinguish each four species from ITS2, matK and rbcL gene, respectively. The sequence differences at the corresponding positions were available genetic marker nucleotides to discriminate the correct species among analyzed four species. These results indicated that comparative analysis of ITS2, matK and rbcL genes were useful genetic markers to authenticate Patriniae Radix. Conclusions : The marker nucleotides enough to distinguish P. scabiosaefolia, P. villosa, P. saniculifolia, and P. rupestris, were obtained at 22 SNP marker nucleotides from ITS2 and matK DNA barcode sequences, but they were confirmed at 12 SNP marker nucleotides from rbcL. These differences could be used to authenticate Patriniae Radix from its adulterants as well as discriminating each four species.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.26 no.3
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• pp.238-247
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• 2021

Diatoms are a type of microalgae which inhabits a wide variety of environments as one of the most important primary producers of freshwater and marine ecosystems. They have been widely used as bioindicators which represent the environmental characteristics, thus proper quality control of diatom data is very important to ensure for the researches from many scientists from various different regions to have scientific unity and objectivity. At present, diatom data analysis is primarily based both on morphological features and DNA sequences of given species. Challenge for the morphology-based analysis of diatoms is the consistent species identification among different taxonomists who interpret diatom community, while challenge for the molecular analysis of diatoms to secure reliable reference data. In the present study, we have reviewed the current status of data quality control of diatom analysis in Korea as well as the world. Finally, suggestions for the better data quality control for Korean marine diatoms have been also made.

• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.274-285
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• 2007

The subfamily Chrominae of damselfishes (Teleostei: Pomacentridae) includes the genus Chromis and Dascyllus. They are found throughout the tropical oceans and form a major component of coral reef communities. There are 5 species of the Chrominae currently recognized in Korea. This study was conducted to infer phylogenetic position of two Korean Chromis species and one Dascyllus species within general category of their each genus in worldwide level. This study also includes one species of Japanese Dascyllus. In the phylogenetic analysis, the Japanese D. aruanus grouped with D. aruanus previously reported from French Polynesia. Korean Chromis fumea grouped with Australian C. nitida and the p-distance value between the two species is relatively very low (0.047). Korean C. notatus grouped together with C. flavomaculata (New Caledonia). In the sequence analysis of some Korean and Japanese damselfishes, there was no sequence variation between D. melanurus (Jeju, Korea) and D. melanurus (Indo-Pacific), but the sequences of the two populations were different in only one nucleotide sites from that of D. melanurus in Indonesian Archipelago. The sequences of Dascyllus aruanus (Japan) were different in two nucleotide sites from it in French Polynesia. There were high difference between the sequences of two Korean species, Chromis fumea and Korean C. notatus. The variations among mitochondrial cytochrome b sequences indicate that the gene sequence could be used as DNA barcode for identification of local populations of D. aruaus and D. melanurus as well as species level.

• Journal of Korean Society of Forest Science
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• v.108 no.2
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• pp.200-207
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• 2019

The aim of this study was to provide the basic information necessary to identify Korean fir (Abies holophylla), momi fir (A. firma), and Nikko fir (A. homolepis), and other fir trees planted in South Korea that are protected by law. Analysis of the morphological characteristics of the needles from each sample was investigated. The shape of the needle-leaf tip from A. holophylla was acute, whereas that from A. firma and A. homolepis was emarginate and that from the protected fir trees was obtuse. The number of stomata on the needles was not significantly different between A. holophylla and A. firma, and the number of stomata on the needles from A. homolepis and the protected fir trees were highly similar. In addition, the genetic differences among the Abies species were analyzed using the sequences of five chloroplast DNA regions-matK, atpF-atpH, rpoC2-rps2, rpoC1, and psbA-trnH.The atpF-atpH and psbA-trnH regions were useful for discriminating A. firma from the other species, but there were no differences among A. holophylla, A. homolepis, and the protected fir trees. The same chloroplast sequences were found in both A. holophylla and A. homolepis, which suggests that additional genetic studies might be necessary to identify the Abies species planted in both South Korea and Japan.

• Horticultural Science & Technology
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• v.35 no.3
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• pp.344-360
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• 2017

Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

• The Korean Journal of Mycology
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• v.47 no.3
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• pp.241-247
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• 2019

In 2017, we collected a wood-decay fungus growing on a dead and decaying herbaceous plant (Reynoutria sachalinensis (F. Schmidt) Nakai) in Dokdo, the far-eastern island of South Korea. Morphologically, this species is characterized by resupinate, coriaceous to corky basidiocarps, poroid hymenophores, pseudodimitic hyphal system, and ellipsoid basidiospores. Based on morphological observation and internal transcribed spacer of ribosomal DNA, the fungus was identified as Xylodon flaviporus (Berk. & M.A. Curtis ex Cooke) Riebesehl & Langer. It is only the second macrofungal species reported from Dokdo, and R. sachalinensis is the first herbaceous plant to be identified as a host for X. flaviporus.