• Title/Summary/Keyword: 분자 성판별

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Development of Specific SNP Molecular Marker from Thistle in the DNA Sequences of Chloroplast TrnL-F and Matk Region Using HRM Analysis (엉겅퀴의 엽록체 TrnL-F와 Matk 영역 염기서열의 HRM 분석을 통한 특이적 SNP 분자마커의 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Kim, Yun-Hee
    • Journal of Life Science
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    • v.29 no.5
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    • pp.524-529
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    • 2019
  • Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, cosmetics and pharmaceutical industries. However, their phylogenetic origins and names are different from each country and quite often they are mixed each other resulting in the confusion for consumers. Particularly when they are very similar based on their morphological characteristics and distributed, it is extremely difficult to differentiate their origins even by specialists. Therefore, identification of each plant species is important for standardizing herbal medicine. Thistle is a medicinal and perennial plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. Although thistle is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish from other similar species from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific thistle species via high resolution melting (HRM) curve analyses. We performed molecular authentication of four different kinds of thistle species from different regions using DNA sequences in the trnL-F and matK chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific thistle species from different country.

A Set of Allele-specific Markers Linked to L Locus Resistant to Tobamovirus in Capsicum spp. (고추의 Tobamovirus 저항성 L 유전자좌와 연관된 대립유전자 특이적인 마커 세트)

  • Lee, Jun-Dae;Han, Jung-Heon;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.30 no.3
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    • pp.286-293
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    • 2012
  • The resistance to Tobamovirus in Capsicum spp. has been known to be controlled by five different alleles ($L^0$, $L^1$, $L^2$, $L^3$, and $L^4$) of L locus on the telomere of long arm of pepper chromosome 11. To develop a set of molecular markers differentiating all the alleles of L locus, we used five pepper differential hosts including Capsicum annuum Early California Wonder (ECW, $L^0L^0$), C. annuum Tisana ($L^1L^1$), C. annuum Criollo de Morelos 334 (CM334, $L^2L^2$), Capsicum chinense PI 159236 ($L^3L^3$), and Capsicum chacoense PI 260429 ($L^4L^4$). Developing a series of CAPS or SCAR markers specifically linked to the alleles was allowed by the sequence comparison of PCR amplicons of the $L^3$-linked markers (189D23M, A339, and 253A1R) and BAC sequences (FJ597539 and FJ597541) in the pepper differentials. Genotypes deduced by these markers in 48 out of 53 $F_1$ hybrids of commercial pepper varieties were consistent with their phenotypes by bioassay using Tobamovirus pathotypes ($P_0$, $P_1$, and $P_{1,2$). Consequently, these markers can be useful to differentiate L alleles and for breeding Tobamovirus resistance in pepper with marker-assisted selection.

Development of Selectable Marker of High Oleate Trait in Peanut (Arachis hypogaea L.) (땅콩에서 고 올레인산 형질관련 분자마커의 선발)

  • Yang, Kiwoung;Pae, Suk-Bok;Park, Chang-Hwan;Lee, Myoung Hee;Jung, Chan-Sik;Son, Jeong-Hee;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.507-514
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    • 2010
  • Peanut(Arachis hypogaea L.) is one of the major oilseed crops. The peanut oil consists of palmitic, oleic and linoleic acids, which are present at levels of 10%, 36-67% and 15-43%, respectively. High oleate mutant of peanut F435 contains 80% oleate and as little as 2% linoleate in seed oil. Previous study indicated that delta 12 fatty acid desaturase is a major enzyme controlling the oleate content in seeds of oilseed crops. F435 sequence alignment of their coding regions disclosed that an extra A(adenine) was inserted at the position +2,823 bp of delta 12 fatty acid desaturase gene. This study was to develop molecular marker (SNP marker) co-segregating with the high oleate trait. Chopyeong ${\times}$ F435 $F_2$ 41 population were investigated using molecular marker and fatty acid assay (NIR and gas chromatography). Finally, this marker segregates Chopyeong type 26 lines, heterotype 9 lines and F435 type 6 lines. These results in our study suggested that SNP marker conform fatty acid assay.

Development and Validation of Real-time PCR to Determine Branchiostegus japonicus and B. albus Species Based on Mitochondrial DNA (Real-time PCR 분석법을 이용한 옥돔과 옥두어의 종 판별법 개발)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1331-1339
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    • 2017
  • DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.

DNA barcoding of Raptor carcass collected in the Paju city, Korea (파주시에서 수집한 폐사체 맹금류의 DNA 바코드 연구)

  • Jin, Seon-Deok;Paik, In-Hwan;Lee, Soo-Young;Han, Gap-Soo;Yu, Jae-Pyoung;Paek, Woon-Kee
    • Korean Journal of Environment and Ecology
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    • v.28 no.5
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    • pp.523-530
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    • 2014
  • One juvenile raptor which was not able to be identified due to its head damage was discovered on a roadside in Janggok-ri, Jori-eup, Paju on 28th June, 2011. The species was identified by DNA barcoding. After polymerase chain reaction (PCR) of the mitochondrial cytochrome c oxidase subunit I gene (COI), we obtained 695 bp sequences. We analyzed the obtained COI sequence with similar sequences from the BOLD systems and BLAST of the NCBI Genbank, and discovered that its sequence showed 100 % similarity values with the one of the five gray-faced buzzards which were previously researched. In addition, it was confirmed to be a female through sex determination using DNA. Such results are important information as it confirms the breeding of the gray-faced buzzards for the first time in 43 years since its breeding was last recorded in 1968, in Paju. Wildlife rescue center needs to work with adjacent consigned registration and preservation institutions when carcass of wild animals is collected or DNA samples are obtained for more accurate both species and sex identification through a systematic management system in the future. Furthermore, the obtained DNA sample of the gray-faced buzzard and COI gene, DNA barcode, could be used as reference standards for similar researches in the future.

Comparison of recently developed classification tools in microarray data analysis (마이크로어레이자료분석에서의 최신 분류방법들의 비교연구)

  • Lee, Jae-Won;Lee, Jeong-Bok;Park, Mi-Ra
    • Proceedings of the Korean Statistical Society Conference
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    • 2002.05a
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    • pp.99-104
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    • 2002
  • cDNA 마이크로어레이자료를 이용한 분류방법은 수많은 유전자의 발현을 동시에 모니터링 할 수 있으므로 특정 질병간의 분자생물학적 변이를 이해하는데 있어 기존의 분류방법보다 신뢰성이 훨씬 높을 것으로 기대되고 있다 최근에 Dudoit et al.(2001)은 cDNA 마이크로어레이를 이용한 유전자발현자료의 분석에 있어 분류를 위한 여러 고전적인 판별분류기법 및 최근에 개발된 기법들을 비교, 평가하였다. 본 논문에서는 Dudoit et al.(2001)에서 다루지 않았던 많은 최신 기법들을 포함하여 인간의 종양 자료뿐만이 아니라 농작물을 포함한 동식물 자료에 적용하여 보다 폭넓은 비교연구를 하였다.

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Development of SNP Molecular Marker for Red-fleshed Color Identification of Peach Genetic Resources (복숭아 유전자원의 적색 과육 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Nam, Eun Young;Cho, Kang Hee;Jun, Ji Hae;Chung, Kyeong Ho
    • Korean Journal of Plant Resources
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    • v.32 no.4
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    • pp.303-311
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    • 2019
  • Various colors of fruit skin and flesh are the most popular commercial criteria for peach classification. In order to breed new red-fleshed peach cultivar, many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection markers to screen seedlings with target traits to increase breeding efficiency. For the comparison of transcription profiles in peach cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red-fleshed peach cultivar, 'Josanghyeoldo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two cultivars were selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNP) were screened from peach EST contigs by high resolution melting (HRM) analysis displayed specific difference between 8 red-fleshed peach cultivars and 24 white-fleshed peach cultivars. All 72 pairs of SNPs were discriminated and the HRM profiles of amplicons were established. In the study reported here, the development of SNP markers for distinguishing between red and white fleshed peach cultivars by HRM analysis offers the opportunity to use DNA markers. This SNP marker could be useful for peach marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach cultivars.

Development of SNP markers for the identification of apple flesh color based on RNA-Seq data (RNA-Seq data를 이용한 사과 과육색 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Park, Seo Jun;Cho, Kang Hee;Lee, Han Chan;Lee, Jung Woo;Choi, In Myung
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.372-378
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    • 2017
  • For comparison of the transcription profiles in apple (Malus domestica L.) cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red flesh apple cultivar, 'Redfield' and white flesh apple cultivar, 'Granny Smith' were investigated by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the red flesh apple cultivar and white flesh apple cultivar were selected for nucleotide sequence determination and homology searches. High resolution melting (HRM) technique measures temperature induced strand separation of short PCR amplicons, and is able to detect variation as small as one base difference between red flesh apple cultivars and white flesh apple cultivars. We applied high resolution melting (HRM) analysis to discover single nucleotide polymorphisms (SNP) based on the predicted SNP information derived from the apple EST database. All 103 pairs of SNPs were discriminated, and the HRM profiles of amplicons were established. Putative SNPs were screened from the apple EST contigs by HRM analysis displayed specific difference between 10 red flesh apple cultivars and 11 white flesh apple cultivars. In this study, we report an efficient method to develop SNP markers from an EST database with HRM analysis in apple. These SNP markers could be useful for apple marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in apple cultivars.

Occurrence of Powdery Mildew Caused by New Race 2F of Podosphaera xanthii on Cucumber in Korea (Podosphaera xanthii의 새로운 Race 2F에 의한 오이 흰가루병 국내 발병 보고)

  • Kim, Young-Ah;Jeong, A-Ram;Jang, Min;Park, Chang-Jin
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.183-189
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    • 2020
  • Powdery mildew is a common and serious disease of the Cucurbitaceae including cucumber (Cucumis sativus) in most areas of the world. To identify causal agents of the powdery mildew and their physiological race(s), we collected cucumber leaves displaying typical symptoms of powdery mildew from different locations in Korea. Based on morphological and molecular characteristics, all powdery mildew isolates were identified as an obligate biotrophic fungal pathogen, Podosphaera xanthii. After inoculation at melon (Cucumis melo) differentials to identify the fungal race(s), P. xanthii isolate MI180427 and IC190611 were identified as race 1 which has been repeatedly reported as dominant race in Korea. However, another isolate SE180328 produced different disease response in the tested differentials, being identified as race 2 which has not been reported in Korea. To confirm the race of SE180328, we inoculated additional melon differentials and determined the isolate as race 2F that is the prevalent race of powdery mildew in Beijing, China. Report of this new race 2F in Korea will be helpful for future breeding programs to develop resistant varieties to this race.

Quantitative Structure-Toxicity Relationships (QSTRs) of Fungicidal Phenylthionocarbamate Derivatives (살균성, Phenylthionocarbamate 유도체들의 정량적인 구조와 독성과의 관계)

  • Sung, Nack-Do;Yang, Sook-Young;Park, Kwaun-Yong
    • Korean Journal of Agricultural Science
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    • v.28 no.1
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    • pp.33-40
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    • 2001
  • The authors attempted to derive a comprehensive quantitative structure-toxicity relationships (QSTRs) between various physicochemical parameters of phenyl substituents in fungicidal phenylthionocarbamate derivatives and toxicity evaluated using TOPKAT calculation. On the basis of this approach we made preditions for toxicity values for not yet tested substances with respect to these systems. The results suggested that the optimal values, $(B_2)_{opt.}=1.54_{\AA}$(Ames mutagenicity), $(R)_{opt.}=0.16$ (car-cinogenicity of male rat), $(\pi)_{opt.)=0.16$ (carcinogenicity of male mouse), $({\varepsilon}LOMO)_{opt}=-0.52e.v.$ ($LD_{50}$ of rat oral), $(B_3){opt.}=1.54_{\AA}$(chronic LOAEU), $(logP)_{opt.}=4.25$ ($LC_{50}$ of Fathead minnow) and $({\sigma})_{opt}=-0.68$ ($EC_{50}$ of Daphnia magna) of phenyl substituents were strongly correlated with the acute and chronic toxicities.

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