• Title/Summary/Keyword: Genetic Similarity

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Molecular Phylogeny of Chattonella (Raphidophyceae) Species from Deungnyang Bay, Korea Using Single-Cell PCR (Single-cell PCR을 이용하여 분석한 득량만 Chattonella 종 (Raphidophyceae)의 분자계통학적 특성)

  • Kim, Jin Joo;Song, Seon Yeung;Park, Tae Gyu
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.24 no.7
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    • pp.967-972
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    • 2018
  • The genus Chattonella belonging to the class raphidophyceae, is a harmful algal bloom species. Recently, its occurrence has been increasing and expanding along the Korean coast. Species identification of the genus Chattonella only by morphological observation is difficult due to the lack of rigid cell walls. In this study, the morphological characteristics and genetic affinity of Chattonella sp. isolated from Deungnyang Bay in 2017 were examined. We carried out single-cell isolation from field samples then sequenced three different areas using the single-cell PCR method: 1) parts of ribosomal operon, the large subunit (LSU) of the rDNA, 2) the chloroplast-encoded subunit psaA of Photosystem I, and 3) rbcL encoding the large subunit of the Rubisco gene. The cells were morphologically very similar to the general genus Chattonella ($74.0{\pm}10.1{\mu}m$ in length, $33.1{\pm}3.6{\mu}m$ in width). The three partial gene sequences were insufficient to justify distinction at the species rank. However, they clustered at 99-100 % sequence similarity with C. marina, C. marina var. antiqua and C. marina var. ovata.

Isolation and identification of tick-borne pathogens in hard ticks collected in Daejeon (대전 주택가 산책로 진드기의 인수공통전염병 병원체 감염실태 조사)

  • Han, So-young;Sung, Sun-hye;Seo, Jin-woo;Kim, Jong-ho;Lee, Seok-ju;Yoo, Sang-sik
    • Korean Journal of Veterinary Service
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    • v.44 no.2
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    • pp.93-102
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    • 2021
  • In this study, a total of 9,449 hard ticks were collected once a month from April to October 2020 from a neighborhood park in Daejeon by flagging & dragging method and CO2 manned trap method. The collected ticks were classified according to the Yamagutsi search table using a stereoscopic microscope and molecular biological analysis of four pathogens (SFTSV, Anaplasma spp., Ehrlichia spp., Borrellia spp.). As a result of the study, Haemaphysalis longicornis were collected the most in all areas of the five boroughs at a rate of 82 to 96 percent, while adults were collected the most in May to July, nymphs were collected the most in April to June, and larvae from August to October at a rate of 78 percent to 98 percent. In pathogens, three cases of SFTSV were detected, showing a minimum infection rate (MIR) of 0.46%, while Anaplasma spp. and Ehrlichia spp. were detected one each, with 0.15% and Borrelia spp. with a minimum infection rate of 0.46%. The detected SFTSV showed 99.9% homogeneity with the KF781490 detected in Cheongwon-gun, Chungbuk Province, Anaplasma spp. showed 99.0% homogeneity with JN990105 detected in China, and Erhlichia spp. showed 98.9% genetic similarity with U96436 separated from the U.S. In this study, the distribution status and pathogen infection rate of the hard ticks in the Daejeon area are analyzed and provided as basic data for the prevention of the hard tick-borne infectious disease.

Development of ITS sequence based SCAR marker and multiplex-SCAR assay for the rapid authentication of Tetrapanacis Medulla and Akebiae Caulis (통초(通草), 목통(木通) 신속 감별용 ITS 염기서열 기반 SCAR 마커 및 Multiplex-SCAR 분석법 개발)

  • Noh, Pureum;Kim, Wook Jin;Park, Inkyu;Yang, Sungyu;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.36 no.1
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    • pp.9-17
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    • 2021
  • Objectives : Tetrapanacis Medulla and Akebiae Caulis are one of the most frequently adulterated herbal medicines because of their confusability of terms in the ancient writings and the similarity of morphological features of dried herbal products. The major adulterant is Aristolochia manshuriensis (Guanmutong) which has a serious safety concern with its toxicity. To ensure the safety and quality of the two herbal medicines, it is necessary to discriminate the toxic adulterant from authentic species. The aim of this study is to develop SCAR markers and to establish the multiplex-SCAR assay for discrimination of four plant species related to Tetrapanacis Medulla and Akebiae Caulis. Methods : ITS regions of fifteen samples of four species (Tetrapanax papyrifer, Fatsia japonica, Aristolochia manshuriensis, and Akebia quinata) collected from different sites were amplified and sequenced. Fifteen obtained ITS sequences were aligned and analysed for the detection of species-specific sequence variations. The SCAR markers were designed based on the sequence alignments and then, multiplex-SCAR assay enhancing rapidity was optimized. Results : ITS sequences clearly distinguished the four species at the species level. The developed SCAR markers and multiplex-SCAR assay were successfully discriminated four species and detected the adulteration of commercial product samples by comparison of the amplified DNA fragment sizes. Conclusions : These SCAR markers and multiplex-SCAR assay are a rapid, simple, and reliable method to identify the authentic Tetrapanacis Medulla and Akebiae Caulis from adulterants. These genetic tools will be useful to ensure the safety and to standardize the quality of the two herbal medicines.

Comparison of Angelica Species Roots Using Taste Sensor and DNA Sequencing Analysis (미각센서와 DNA 염기서열을 이용한 당귀류 비교)

  • Kim, Young Hwa;Choi, Goya;Lee, Hye Won;Lee, Gwan Ho;Chae, Seong Wook;Kim, Yun Hee;Lee, Mi Young
    • The Korea Journal of Herbology
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    • v.27 no.6
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    • pp.37-42
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    • 2012
  • Objectives : Angelica Gigantis Radix is prescribed as the root of different Angelica species on the pharmacopoeia in Korea, Japan and China. Chemical components and their biological activities were also different according to their species. A study for the development of simple method to compare Angelica roots was needed. In order to classify them, the methods such as DNA sequencing analysis and taste sensor were applied to three Angelica species like Angelica gigas, Angelica acutiloba and Angelica sinensis. Methods : PCR amplification of intergenic transcribed spacer (ITS) region was performed using ITS1 and ITS4 primer from nine Angelica roots, and then nucleotide sequence was determined. Taste pattern of samples were measured using the taste-sensing system SA402B equipped with a sensing unit, which consists of artificial lipid membrane sensor probes of anionic bitterness, astringency, saltiness, umami, and cationic bitterness (C00, AE1, CT0, AAE, and AN0, respectively). Results : As a result of comparing the similarity of the ITS region sequences, A. sinensis was discriminated from the others (A. gigas and A. acutiloba). Equally this genetic result, A. gigas and A. acutiloba showed similar taste pattern as compared to A. sinensis. Sourness, bitterness, aftertaste of bitterness, astringency, and aftertaste of astringency of A. sinensis were significantly high as compared with A. gigas and A. acutiloba. In contrast, richness was significantly low. Conclusions : These taste pattern can be used as a way of comparison of Angelica species and this technic could be applied to establish a taste pattern marker for standardization of herbs in various purposes.

Sinomonas terrae sp. nov., Isolated from an Agricultural Soil

  • Hyosun Lee;Ji Yeon Han;Dong-Uk Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.909-914
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    • 2023
  • While searching for the bacteria which are responsible for degradation of pesticide in soybean field soil, a novel bacterial strain, designated 5-5T, was isolated. The cells of the strain were Gram-staining-positive, aerobic and non-motile rods. Growth occurred at 10-42℃ (optimum, 30℃), pH 5.5-9.0 (optimum, pH 7.0-7.5), and 0-2% (w/v) NaCl (optimum, 1%). The predominant fatty acids were C15:0 anteiso, C17:0 anteiso, and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The predominant menaquinone was MK-9 (H2). Diphosphatidylglycerol, glycolipids, phosphatidylinositol, and phosphatidylglycerol were the major polar lipids. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain 5-5T is a member of the genus Sinomonas and its closest relative is Sinomonas humi MUSC 117T, sharing a genetic similarity of 98.4%. The draft genome of strain 5-5T was 4,727,205 bp long with an N50 contig of 4,464,284 bp. Genomic DNA G+C content of strain 5-5T was68.0 mol%. The average nucleotide identity (ANI) values between strain 5-5T and its closest strains S. humi MUSC 117T and S. susongensis A31T were 87.0, and 84.3 % respectively. In silico DNA-DNA hybridization values between strain 5-5T and its closest strains S. humi MUSC 117T and S. susongensis A31T were 32.5% and 27.9% respectively. Based on the ANI and in silico DNA-DNA hybridization analyses, the 5-5T strain was considered as novel species belonging to the genus Sinomonas. On the basis of the results from phenotypic, genotypic and chemotaxonomic analyses, strain 5-5T represents a novel speciesof the genus Sinomonas, for which the name Sinomonas terrae sp. nov. is proposed. The type strain is 5-5T (=KCTC 49650T =NBRC 115790T).

Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

Variations in Ecological Niche of Quercus variabilis and Quercus acutissima Leaf Morphological Characters in Response to Moisture and Nutrient Gradient Treatments under Climate Change Conditions (기후변화 조건에서 수분구배 및 영양소 구배에 따른 굴참나무와 상수리나무 잎 형태적 특성의 생태지위 변화)

  • Park, Yeo-Bin;Kim, Eui-Joo;Park, Jae-Hoon;Kim, Yoon-Seo;Park, Ji-Won;Lee, Jung-Min;You, Young-Han
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.27 no.2
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    • pp.43-53
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    • 2024
  • This study attempted to elucidate the ecological niches and influencing environmental factors of Quercus variabilis and Quercus acutissima, which are representative deciduous broad-leaved trees in Korean forests, taxonomically close and genetically similar, under climate change conditions. Under climate change conditions induced by increased CO2 and temperature, soil moisture and nutrient environments were manipulated in four gradients. At the end of the growing, plants were harvested to measure growth responses, calculate ecological niches, and compare them with those of the control. Eperimental plants were grown for 180 days in a glass greenhouse designed with four gradients each for soil moisture and nutrient environments under climate change conditions induced by increased CO2 and temperature. After harvesting, growth responses of leaf traits were measured, ecological niches were calculated, and these were compared with those of the control groups. Furthermore, the responses of the two species' populations were interpreted using principal component analysis(PCA) based on leaf trait measurements. As a result, under climate change conditions, the ecological niche breadth for moisture environment was broader for Quercus variabilis than Quercus acutissima, whereas for the nutrient environment, Quercus acutissima exhibited a broader niche breadth than Quercus variabilis. And the rate of change in ecological niche breadth due to climate change decreased for Quercus variabilis in both moisture and nutrient environments, while for Quercus acutissima, it increased in the moisture environment but decreased in the nutrient environment. Additionally, in terms of group responses, both Quercus variabilis and Quercus acutissima expanded their ecological niches under climate change conditions in both soil moisture and nutrient conditions, with Quercus acutissima exhibiting a broader niche than Quercus variabilis under nutrient conditions. These results indicate that the changes in leaf morphological characteristics and the responses of individuals reflecting them vary not only under climate change conditions but also depending on environmental factors.

Fruiting body development and genetic analysis of somatic hybrids by protoplast fusion in edible fungi (식용버섯의 원형질체 융합체의 자실체 발생 및 유전분석)

  • Yoo, Young Bok;Kong, Won Sik;Oh, Se Jong;Jhune, Chang Sung;Shin, Pyung Gyun;Kim, Beom Gi;Kim, Gyu Hyun;Park, Minsun;Min, Byung Re
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.115-126
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    • 2004
  • Somatic hybrids of inter-compatible and inter-incompatible strains were obtained by protoplast fusion. The fusion products between compatible strains, Pleurotus ostreatus and P. florida, formed heterokaryons, while fusants between incompatible strains such as P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, and P. ostreatus + Flammulina velutipes formed synkaryons that retained genes from both parents. The heterokaryons showed the same level of basidioma development. In contrast, the synkaryons showed unique characteristics including clamp connection formation at mitosis, either partner basidioma development, and abnormal segregation and recombination compared with inter-compatible strains. Synkaryons can be classified into homokaryoyic and heterokaryotic type. A comparison of somatic hybrids with compatible and incompatible strains was made using random amplified polymorphic DNA (RAPD) analysis. The heterokaryons between compatible species showed the same level of variability and contained both parental RAPD bands. In contrast, most of the synkaryons between incompatible species showed similarity to those of either parental bands and non-parental RAPD bands. Synkaryons can be classified into microgenome insertion type and macrogenome insertion type. A tetrapolar mating system was found among monospore isolates in somatic hybrids and wild type P. ostreatus. Homokaryons from each somatic hybrid combination were paired with tester homokaryons of the initial wild type of P. ostreatus. The changed mating types were identified in progenies. The pattern of mating type switching in somatic hybrids depends on compatibility of fusion partner. There are several factors related to the mechanism of clamp connection formation and fruiting body development of synkaryons. Of these,the major factor may be associated with self-fertility and mating type switching such as homokaryotic fruiting of wild type P. ostreatus. This review will discuss these aspects.

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Analysis of Potential Toxigenicity and Phylogeny using Target Genes in Aphanizomenon flos-aquae (Cyanophyceae) strains isolated from the Nakdong River (낙동강에서 분리된 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 목표 유전자를 이용한 잠재적 독소 생성능 및 계통학적 분석)

  • Ryu, Hui-Seong;An, Sung-Min;Lim, Chang-Kun;Shin, Ra-Young;Park, Jong-Guen;Lee, Jung-Ho
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.137-147
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    • 2017
  • The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

Molecular Phylogenetic Study of the Endangered Land Snail Satsuma myomphala Based on Metallothionein Gene. (Metallothionein 유전자를 기초로 한 멸종위기 육상 달팽이 Satsuma myomphala (거제외줄달팽이) 의 분자계통학적 연구)

  • Sang, Min Kyu;Kang, Se Won;Hwang, Hee-Ju;Chung, Jong Min;Song, Dae Kwon;Min, Hye Rin;Park, Jie Eun;Ha, Hee Cheol;Lee, Hyun Jun;Hong, Chan Eui;Ahn, Young Mo;Park, So Young;Park, Young-Su;Park, Hong Seog;Han, Yeon Soo;Lee, Jun Sang;Lee, Yong Seok
    • 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.