• Journal of Food Hygiene and Safety
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• v.39 no.3
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• pp.209-220
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• 2024

Cavier is graded as Beluga, Osetra, and Sevruga based on the species of sturgeon (Acipenser sinensis). In this study, we developed an analytical method for determining the grade of black caviar using DNA barcodes and KASP markers. To identify the sturgeon species, ten black caviar samples were collected, and a reference DNA barcode library was developed using five genes (namely, 16S ribosomal RNA, cytochrome b, cytochrome c oxidase subunit I, cytochrome c oxidase subunit II, and NADH dehydrogenase subunit 5 genes). To develop the KASP markers, we selected 11 markers that could distinguish between the five grades of black caviar. As a result, specific markers for each of the targeted caviars were clustered into FAM-positive sections. DNA barcoding and the KASP assay revealed that one Beluga, six Osetra, and three Sevruga were identified among the ten caviar samples. Moreover, we found that the sturgeon species were mislabeled in two products. Here, we aimed to develop a KASP assay based on SNP that allows rapid and easy identification of caviar grade. These methods are expected to contribute to preventing the distribution of illegal products.

• Korean Journal of Poultry Science
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• v.36 no.1
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• pp.85-88
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• 2009

One of the mitochondrial genes, called cytochrome c oxidase I (COI), has been widely used for the species identification (called bio-barcode) in birds. In this study, the bio-barcode has been applied to chicken breeds in Korea whether it also can be used as a molecular marker for breed identification. Data indicated that Korean native chicken has the mixed SNP (single nucleotide polymorphism) patterns between White Leghorn (Layer) and Cornish (Broiler) and ultimately, it can not be used as the marker for breed identification. However, this result indicates the mixed use of the Korean native chicken, since it has been used for dual purpose for producing meat and egg for a long time. In order to use as a marker for species identification, more reliable mitochondrial and/or nuclear DNA markers need to be developed.

• Korean Journal of Environment and Ecology
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• v.32 no.6
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• pp.566-574
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• 2018

The purpose of this study was to identify the phylogenetic relationships of the plants in the Korean genus Suaeda and to find out the molecular markers that could confirm the interspecies relationships in the family tree through molecular phylogenetic studies. We used the nuclear ribosomal DNA ITS and the chloroplast DNA matK, psbA-trnH, and trnL-trnF as the molecular markers. We could not distinguish between S. japonica and S. maritima and between S. maritima and S. australis in the ITS region and could not distinguish between S. japonica and S. australis with the base sequence in the psbA-trnH and trnL-trnF region. However, we analyzed the combinations of four molecular marker regions and confirmed that each of five plant species of the genus Suaeda formed the independent line. Therefore, it is considered that combinations of molecular markers would be useful for the analysis of phylogenetic relationships in the genus Suaeda. Further investigations of the ecological and morphological characteristics would be needed to understand the phylogenetic relationship and lineage diversification in the genus Suaeda.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.15-25
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• 2020

Allium L. is one of the largest genera of the Amaryllidaceae family, with more than 920 species including many economically important species used as vegetables, spices, medicines, or ornamental plants. Currently, DNA barcoding tools are being successfully used for the molecular taxonomy of Allium. A total of 46 Allium species were collected from their native areas, and DNA was extracted using the IBRC DNA extraction kit. We used specific primers to PCR amplify matK. DNA sequences were edited and aligned for homology, and a phylogenetic tree was constructed using the neighbor-joining method. The results show thymine (38.5%) was the most frequent and guanine (13.9%) the least frequent nucleotide. The matK regions of the populations were quite highly conserved, and the amount of C and CT was calculated at 0.162 and 0.26, respectively. Analysis of the nucleotide substitution showed C-T (26.22%) and A-G (8.08%) to have the highest and lowest percent, respectively. The natural selection process dN/dS was 1.16, and the naturality test results were -1.5 for Tajima's D and -1.19 for Fu's Fs. The NJ dendrogram generated three distinct clades: the first contained Allium austroiranicum and A. ampeloprasum; the second contained A. iranshahrii, A. bisotunense, and A. cf assadi; and the third contained A. rubellum and other species. In this study, we tested the utility of the matK region as a DNA barcode for discriminating Allium. species.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.97-97
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• 2019

사삼(沙蔘, Adenophorae Radix)은 "대한민국약전외한약(생약)규격집(KHP)"에 잔대 Adenophora triphylla var. japonica Hara 또는 사삼(당잔대, A. stricta Miq.)의 뿌리로 수재되어 있으나, 형태학적으로 유사한 제니(모시대, A. remotiflorus Miquel), 층층잔대(윤엽사삼, A. tetraphylla (Thunb.) Fisch), 더덕 Codonopsis lanceolata (Sieb. et Zucc.)과 오 혼용 우려가 있어 이들을 구별하기 위한 종 감별법이 필요하다. 본 연구에서는 '사삼'과 오 혼용 우려가 있는 종들을 구별할 수 있는 유전자 마커 개발을 위하여 DNA 바코드로 활용되고 있는 유전자 부위를 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. 또한, 분석한 유전자 부위 중 종간 차이를 확인하기 용이한 matK 구간을 활용해 기원종인 잔대, 당잔대와 형태적으로 유사하여 오 혼용될 우려가 있는 층층잔대, 모시대 및 더덕을 감별 할 수 있는 유전자 마커를 개발하였다. 본 연구를 통해 얻어진 염기서열과 분자 마커는 '사삼'의 품질관리에 유용하게 활용 가능할 것으로 사료된다.

• Genomics & Informatics
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• v.16 no.2
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• pp.22-29
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• 2018

Incorporation of unique barcodes into fission yeast gene deletion collections has enabled the identification of gene functions by growth fitness analysis. For fine tuning, it is important to examine barcode sequences, because mutations arise during strain construction. Out of 8,708 barcodes (4,354 strains) covering 88.5% of all 4,919 open reading frames, 7,734 barcodes (88.8%) were validated as high-fidelity to be inserted at the correct positions by Sanger sequencing. Sequence examination of the 7,734 high-fidelity barcodes revealed that 1,039 barcodes (13.4%) deviated from the original design. In total, 1,284 mutations (mutation rate of 16.6%) exist within the 1,039 mutated barcodes, which is comparable to budding yeast (18%). When the type of mutation was considered, substitutions accounted for 845 mutations (10.9%), deletions accounted for 319 mutations (4.1%), and insertions accounted for 121 mutations (1.6%). Peculiarly, the frequency of substitutions (67.6%) was unexpectedly higher than in budding yeast (~28%) and well above the predicted error of Sanger sequencing (~2%), which might have arisen during the solid-phase oligonucleotide synthesis and PCR amplification of the barcodes during strain construction. When the mutation rate was analyzed by position within 20-mer barcodes using the 1,284 mutations from the 7,734 sequenced barcodes, there was no significant difference between up-tags and down-tags at a given position. The mutation frequency at a given position was similar at most positions, ranging from 0.4% (32/7,734) to 1.1% (82/7,734), except at position 1, which was highest (3.1%), as in budding yeast. Together, well-defined barcode sequences, combined with the next-generation sequencing platform, promise to make the fission yeast gene deletion library a powerful tool for understanding gene function.

• Journal of Plant Biotechnology
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• v.45 no.1
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• pp.9-16
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• 2018

The advent of available DNA barcoding technology has been extensively adopted to assist in the reference to differentiate the origin of various medicinal plants species. However, this technology is still far behind the curve of technological advances to be applied in a practical manner in the market to authenticate the counterfeit components or detect the contamination in the admixtures of medicinal plant species. Recently, a high resolution melting curve analysis technique was combined with the procedure of DNA barcoding (Bar-HRM) to accomplish this purpose. In this review, we tried to summarize the current development and bottleneck of processing related to the Bar-HRM technology for the practical application of medicinal plant species' differentiation in a viable global market. Although several successful results have been reported, there are still many obstacles to be resolved, such as limited number of DNA barcodes and single nucleotide polymorphisms, in particular, only one DNA barcode, internal transcribed sequence (ITS) of ribosomal DNA has been reported in the available nuclear genome. In addition, too few cases have been reported about the identification of counterfeit or contamination with processed medicinal plant products, in particular specifically the case of technology based infusion, jam and jelly products and components in which it is noted that DNA can be thereby degraded during the processing of these products and components.

• 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.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.464-475
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• 2023

In this study, based on an analysis of two DNA barcode markers (cytochrome c oxidase subunit I and cytochrome b genes), we performed species identification and monitored labeling compliance for 50 commercial pufferfish products sold in on-line markets in Korea. Using these barcode sequences as a query for species identification and phylogenetic analysis, we screened the GenBank database. A total of seven pufferfish species (Takifugu chinensis, T. pseudommus, T. xanthopterus, T. alboplumbeus, T. porphyreus, T. vermicularis, and Lagocephalus cheesemanii) were identified and we detected 35 products (70%) that were non-compliant with the corresponding label information. Moreover, the labels on 12 commercial products contained only the general common name (i.e., pufferfish), although not the scientific or Korean names for the 21 edible pufferfish species. Furthermore, the proportion of mislabeled highly processed products (n = 9, 81.8%) was higher than that of simply processed products (n = 26, 66.7%). With respect to the country of origin, the percentage of mislabeled Chinese products (n = 8, 80%) was higher than that of Korean products (n = 26, 66.7%). In addition, the market and dialect names of different pufferfish species were labeled only as Jolbok or Milbok, whereas two non-edible pufferfish species (T. vermicularis and T. pseudommus) were used in six commercial pufferfish products described as JolboK and Gumbok on their labels, which could be attributable to the complex classification system used for pufferfish. These monitoring results highlight the necessity to develop genetic methods that can be used to identify the 21 edible pufferfish species, as well as the need for regulatory monitoring of commercial pufferfish products.

• International Journal of Industrial Entomology and Biomaterials
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• v.15 no.1
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• pp.47-58
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• 2007

The tiny dragonfly, Nannophya pygmaea(Odonata: Libellulidae) is one the smallest dragonflies in the world and listed as a second-degree endangered wild animal and plant in Korea. For the long-term conservation of such endangered species, an investigation on nation-wide genetic magnitude and nature of genetic diversity is required as a part of conservation strategy. We, thus, sequenced a portion of mitochondrial COI gene, corresponding to "DNA Barcode" region(658 bp) from 68 N. pygmaea individuals collected over six habitats in Korea. The sequence data were used to investigate genetic diversity within populations and species, geographic variation within species, phylogeographic relationship among populations, and phylogenetic relationship among haplotypes. Phylogenetic analysis and uncorrected pairwise distance estimate showed overall low genetic diversity within species. Regionally, populations in southern localities such as Gangjin and Gokseong in Jeollanamdo Province showed somewhat higher genetic diversity estimates than those of remaining regions in Korean peninsula. Although geographic populations of N. pygmaea were subdivided into two groups, distance- or region-based geographic partition was not observed.