• 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.

• Korean Journal of Medicinal Crop Science
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• v.26 no.1
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• pp.26-31
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• 2018

Background: Angelica gigas, A. sinensis, and A. acutiloba are commercially important in the herbal medicine market, and among them, A. gigas has the highest economic value and price. However, their similar morphological traits are often used for fraud. Despite their importance in herbal medicine, recognition of the differences between Angelica species is currently inadequate. Methods and Results: A multiplex polymerase chain reaction (PCR) method was developed for direct detection and identification of A. gigas, A. sinensis, and A. acutiloba. The gene for the distinction of species was targeted at ITS in the nucleus and trnC-petN gene in chloroplasts. The optimized multiplex PCR in the present study utilized each Angelica species-specific primer pairs. Each primer pair yielded products of 229 base pairs (bp) for A. gigas, 53 bp for A. sinensis, 170 bp for A. acutiloba. Additionally non-specific PCR products were not detected in similar species by species-specific primers. Conclusions: In the present study, a multiplex-PCR assay, successfully assessed the authenticity of Angelica species (A. gigas, A. sinensis, and A. acutiloba). and whole genome amplification (WGA) was performed after DNA extraction to identify, the species in the product. The detection method of raw materials developed in the present study could be applied to herbal medicine and health functional food management.

• ALGAE
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• v.30 no.3
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• pp.217-222
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• 2015

Pythium species (Pythiales, Oomycetes) are well known as the algal pathogen that causes red rot disease in Pyropia / Porphyra species (Bangiales, Rhodophyta). Accurate species identification of the pathogen is important to finding a scientific solution for the disease and to clarify the host-parasite relationship. In Korea, only Pythium porphyrae has been reported from Pyropia species, with identifications based on culture and genetic analysis of the nuclear internal transcribed spacer (ITS) region. Recent fungal DNA barcoding studies have shown the low taxonomic resolution of the ITS region and suggested the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene as an alternative molecular marker to identify Pythium species. In this study, we applied an analysis of both the ITS and cox1 regions to clarify the taxonomic relationships of Korean Pythium species. From the results, the two closely related Pythium species (P. chondricola and P. porphyrae) showed the same ITS sequence, while the cox1 marker successfully discriminated P. chondricola from P. porphyrae. This is the first report of the presence of P. chondricola from the infected blade of Pyropia yezoensis in Asia. This finding of the algal pathogen provides important information for identifying and determining the distribution of Pythium species. Further studies are also needed to confirm whether P. chondricola and P. porphyrae are coexisting as algal pathogens of Pyropia species in Korea.

• The Journal of the Korean Society for Microbiology
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• v.22 no.2
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• pp.117-123
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• 1987

Previous studies have been performed for the sero-identification of selected species of Bacteroides by immunofluorescence antibody techniques and enzyme-linked immunosorbent assay using species-specific rabbit antisera to B. gingivalis, B. intermedius, and B. melaninogenicus. However, these studies have not commented on the serological cross-reactivity between these 3 species of black- pigmented Bacteroides. For the cross-reactivity study, antisera to B. gingivalis ATCC 33277, B. intermedius ATC C25261 and B. asaccharolyticus ATCC 25260 were raised from rabbits. Preliminary study for observing the cross-reactivity between these species was performed by indirect immunoflourescence technique. Immunoabsorption of the antisera was done with bacterial cells from the other species and the species-specificity of the antisera was conformed by the absence of reactivity with bacterial strains from the other species by indirect immunofluorescence technique and enzyme-linked immunosorbent assay. Three representative unabsorbed antisera cross-reacted strongly with cells from the other species. Especially, anti-B. asaccharolyticus ATCC 25260 antiserum showed a strong cross-reactivity with B. gingivalis ATCC33277. After immunoabsorption of 3 representative antisera with the other species, the cross-reactivity was found only between B. gingivalis ATCC 33277 and B. asaccharolyticus ATCC 25260. Further study would be necessary to clarify the cross-reactivity between important oral black-pigmented Bacteroides from subgingival plaque or bacterial colonies for rapid identification.

• Journal of the Korean Wood Science and Technology
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• v.46 no.3
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• pp.270-277
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• 2018

This study was conducted to identify wood species of two printing woodblocks either from the Park clan's documentary or Ji-dang documentary of Songok clan of the Milseong Park, Songok, Gyeongju, Korea. Eighty-eight woodblocks out of the total 282 woodblocks were randomly selected to compare anatomical features for the identification of wood species, using a light microscope. As a result, seven wood species were identified, and all of them were diffuse-porous hardwood species. The most significant portion, i.e., 39.8% of wood species was Carpinus laxiflora Blume. Then, Pyrus pyrifolia Nakai, Acer mono Maxim, Prunus sargentii Rehder, Tilia amurensis Rupr, Diospyros kaki Thunb, and Betula costata Trautv was 25.0%, 15.9%, 10.2%, 3.4%, 3.4% and 2.3%, respectively, indicating that all diffuse-porous hardwood species had been used for the woodblocks. It was believed that diffuse-porous hardwoods had been used because they provided an easy of engraving complex Chinese letters, of acquiring these wood species in Gyeongju areas, and a high resistance to repeated printing.

• Mycobiology
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• v.48 no.6
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• pp.476-483
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• 2020

The genus Pholiota (Strophariaceae, Basidiomycota) is made up of wood-rotting saprotrophic mushrooms characterized by a yellow or brown pileus with scales and/or slimy, and by a brownish smooth spore with a germ pore. However, these features are not enough to distinguish its species, or separate the genus Pholiota from other brown-spored wood-rotting genera such as Hypholoma and Stropharia. Although internal transcribed spacer (ITS) sequencebased identification has improved identification accuracy for species of Pholiota, most Pholiota species in Korea are reported based on morphological features. To evaluate the taxonomy of Pholiota species, we investigated 62 specimens collected from 1999 to 2019 in Korea using ITS sequence analysis and morphological observation. Twelve of the 16 recorded Pholiota species in Korea were identified. While eight species were clearly separated, the ITS analysis did not distinguish three in the Pholiota adiposa complex. Therefore, further investigation is required to distinguish these three species. ITS sequences deposited in GenBank confirm that P. highlandensis exists in Korea. The presence of the other four Pholiota species could not be confirmed through specimens or sequence information in GenBank. A taxonomic key and the ITS sequence data for Korean Pholiota species are included and can be good baselines for further research on Pholiota taxonomy and diversity.

• Journal of Practical Agriculture & Fisheries Research
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• v.24 no.3
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• pp.5-14
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• 2022

Trichoderma is known as pathogen caused serious green mold disease on commercial production. T. pleuroti and T. pleuroticola were common species in various mushroom media. Many strains of T. pleuroti, known as aggressive species causing major economic losses in Korea, showed benomyl resistance. Accurate identification and detection of Trichoderma species associated with oyster mushrooms is very important for disease control. We developed species-specific primers for T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride based on species-specific fragments isolated from amplified fragment length polymorphism analysis. PCR products corresponding to the predicted fragment of 500bp, 230bp, 180bp, and 410bp were amplified from T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride, respectively. Multiplex PCR assay using species-specific primers quickly and accurately identified and detected T. pleuroti from mushroom media in which various species co-exist. Our results can be useful for the effective control of mushroom disease.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.159-163
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• 2008

Accurate identification of pine wood nematode, Bursaphelenchus xylophilus is a prerequisite to diagnose the pine wilt disease. However, a fungivorous nematode, B. mucronatus is highly similar to B. xylophilus and it is difficult to differentiate these two species by morphological features. A molecular diagnosis method, ITSRFLP was applied for the identification of B. xylophilus and B. mucronatus from Korea. Genomic DNA was extracted from a single individual nematode and ITS DNA was amplified by PCR. The size of PCR product was approximately 900bp and the sequence data were obtained after cloning. Amplified ITS was digested by 5 different restriction enzymes (Rsa I, Hae III, Msp I, Hinf I, and Alu I) and provided a discriminatory profile for B. xylophilus and B. mucronatus. Besides, B. mucro- natus was determined to have 2 different genotypes, East Asian type and European type also clearly separated by Rsa I and Hae III digestion. European type of B. mucronatus is recently collected from Pinus koraiensis and has not been reported before. ITS sequnce data were analyzed by Restriction Mapper program and the result supported ITS-RFLP pattern. These data indicated that PCRRFLP method is an accurate and simple way for identification of Bursaphelenchus species.

• Food Science of Animal Resources
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• v.27 no.2
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• pp.209-215
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• 2007

In order to develop a sensitive and reliable method for the species-specific molecular markers, PCR-RFLP assay of the mitochondrial DNA(mt DNA) 12S rRNA gene was exploited for the identification of the origin of animal meat species including cattle, pig, sheep, goat, horse, deer, chicken, duck and turkey. A specific primer pairs were designed, based on the nucleotide sequences of mt 12S rRNA gene, for the amplification of the highly conserved region in the gene of the animal species using PCR-RFLP technique. mt DNA was isolated from meat samples followed by DNA amplification using PCR with the specific primers. PCR amplification produced an approximately 455 bp fragment in each of these animal meats. To distinguish pleat species, the PCR amplicons were digested with restriction endonucleases Tsp5091 and MboI, respectively, which generates distinct RFLP profiles. The DNA profiles digested with Tsp5091 allowed the clear discrimination in the mammalian meat species and the DNA profiles digested with MboI in poultry meat species. Therefore, the PCR-RFLP profiles of mt 12S rRNA gene could be very useful to identify the origin of the raw materials in the raw meats as well as the processed meat products.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2005.04a
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• pp.40.1-40.1
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• 2005