• Title/Summary/Keyword: 분자 마커

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Construction of a Genetic Linkage Map in Radish(Raphanus sativus L.) Using RAPD Markers (RAPD 마커를 이용한 무의 유전자지도 작성)

  • Ahn, Choon-Hee;Choi, Su-Ryun;Lim, Yong-Pyo;Chung, Hae-Joon;Yae, Byeong-Woo;Yoon, Wha-Mo
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.151-159
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    • 2002
  • Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n=9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

Development of DNA Markers for Trehalose Synthesis Genes in Brassica rapa L. (배추 trehalose 합성 유전자와 연관된 DNA 마커 개발)

  • Jeong, Ye-Sol;Lim, Yong-Pyo;Hur, Yoon-kang;Chung, Sang-Min
    • Journal of Life Science
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    • v.19 no.5
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    • pp.639-643
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    • 2009
  • High temperature stress might affect the yield and quality of Chinese cabbage. In order to develop cultivars resistant to high temperature stress, we developed polymorphic DNA markers for trehalose synthesis genes related to abiotic stress resistance. A total of 28 Brassica rapa ESTs homologous to trehalose synthesis genes of Arabidopsis were found from the NCBI database. The polymorphic DNA sequences were searched between Chinese cabbages - Chiifu, which is relatively susceptible to high temperature stress, and Kenshin, which is tolerant to high temperature stress. Among the 28 ESTs, we found 10 ESTs that have either insertion/deletion and/or single nucleotide polymorphism between the two cultivars. Those polymorphic sites were then targeted for the development of 10 PCR based markers. These molecular markers related to trehalose genes could be used not only to test their relationship with abiotic stress resistance in Chinese cabbage, but also the development of abiotic stress resistant cultivars using MAS.

Isolation and characterization of microsatellite markers for Hydrangea luteovenosa (Hydrangeaceae), an endangered species in Korea (한국 제주도에 자생하는 멸종위기종 성널수국(수국과)의 microsatellite 분자마커 개발)

  • Ito, Takuya;Kaneko, Shingo;Yokogawa, Masashi;Song, Gwan-Pil;Choi, Hyeok-Jae;Isagi, Yuji
    • Korean Journal of Plant Taxonomy
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    • v.43 no.1
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    • pp.30-33
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    • 2013
  • Hydrangea luteovenosa is a critically endangered plant species of Jeju Island in Korea, though it is widely distributed in western Japan. We isolated and characterized five microsatellite loci in this species. The number of alleles ranged from 3 to 27, observed heterozygosity from 0.27 to 0.86, and expected heterozygosity from 0.34 to 0.91. The markers described here will be useful for investigating the genetic diversity, genetic structure, and gene flow of H. luteovenosa, and the genetic findings would contribute to the establishment of effective conservation measures for this species in Korea.

Stress Evaluation to Heavy Metal Exposure using Molecular Marker in Chironomus riparius (분자지표 유전자 발현을 통한 Chironomus riparius 중금속 노출 스트레스 평가)

  • Kim, Won-Seok;Park, Kiyun;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.53 no.2
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    • pp.165-172
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    • 2020
  • Heavy metals are common pollutants in the freshwater environment and have toxicological effect in habitat organisms. The heavy metals highly accumulated in sediment and organism, and observed various physiological responses. In this study, we investigated the molecular response to heavy metal toxicity (Al, Aluminum; Cr, Chromium; Cu, copper; Mn, Manganese; Zn, Zinc) through expression of heat shock protein 40, 70, 90 (HSP40, 70, 90), cytochrome 450 (CYP450), Glutathione S-transferase (GST) and Serine-type endopeptidase (SP). HSPs showed up-regulation in Cu and Zn exposures. Furthermore, HSPs expression in treated groups tended to be higher than the control group. The tendency of CYP450 and GST mRNA expression was higher for Cr and Cu than for other exposure group. The expression of SP gene was low at Al exposure and other group were measured to be similar to control. These results suggest that heavy metal toxicity in freshwater ecosystem may affect physiological and molecular process. Also, the comprehensive gene expression in the aquatic midge Chironomus riparius give useful information to potential molecular biomarkers for assessing heavy metal toxicity.

SSR Marker Related to Major Characteristics Affected Kernel Quality in Waxy Corn Inbred Lines (찰옥수수 자식계통의 주요 품질특성과 관련된 SSR마커)

  • Jung, Tae-Wook;Moon, Hyeon-Gui;Son, Beom-Young;Kim, Sun-Lim;Kim, Soon-Kwon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.185-192
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    • 2006
  • This experiment was conducted to assess genetic diversity of waxy corn inbred lines and to identify SSR markers related to major characteristics affected kernel quality for improving waxy corn $F_1$ hybrid with good quality. Diversity of 64 waxy com inbred lines was evaluated using 30 microsatellite markers. The 30 microsatellite markers representing 30 loci in the maize genome detected polymorphisms among the 64 inbred lines and revealed 225 alleles with a mean of 7.5 alleles per primer. The polymorphism Information content (PIC) value ranged from 0.14 to 0.87, with an average of 0.69. Based on Nei's genetic distances, the 64 inbred lines were classified into 9 groups by the cluster analysis. The group I included 26 inbred lines (41%), other groups included 3 to 9 inbred lines. One-way analysis of variance was conducted to identify significant relationship between individual markers and major characteristics that affect kernel quality. The analysis showed that umc1019 was related to amylopectin and crude protein content, me 1020 to amylopectin content and peak viscosity, and bnlg1537 to 100-kernel weight, kernel length, and kernel width.

A Gene-based dCAPS Marker for Selecting old-gold-crimson (ogc) Fruit Color Mutation in Tomato (토마토 과색 돌연변이 유전자(old-gold-crimson) 선발을 위한 dCAPS 분자표지 개발)

  • Park, Young-Hoon;Lee, Yong-Jae;Kang, Jum-Soon;Choi, Young-Whan;Son, Beung-Gu
    • Journal of Life Science
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    • v.19 no.1
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    • pp.152-155
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    • 2009
  • The old-gold-crimson ($og^c$) fruit color mutation produces deep red tomato fruit with high lycopene content. age is a null mutation allele of lycopene-${\beta}$-cyclase (Crt-b) gene (B locus) that converts lycopene to ${\beta}$-carotene in the cartenoid biosynthesis pathway in tomato. Breeding of high lycopene tomato cultivars can be accelerated by marker-assisted selection (MAS) for introgression of $og^c$ allele by using a gene-based DNA marker. In order to develop a marker, single nucleotide deletion of adenine(A) with. in a poly-A repeat that has been known to be responsible for frame-shift mutation of $og^c$ was confirmed by resequencing mutant allele and wild-type allele at B locus of several tomato lines. For allele discrimination and detection of $og^c$, derived CAPS (dCAPS) approach was used by designing a primer that artificially introduced restriction enzyme recognition site of Hin fI in PCR products from $og^c$ allele. This dCAPS marker is co-dominant gene-based PCR marker that can be efficiently used for MAS breeding program aiming the development of high lycopene tomato.

An Introduction to Microsatellite Development and Analysis (Microsatellite 개발 및 분석법에 대한 소개)

  • Yun Young-Eun;Yu Jeong-Nam;Lee Byoung-Yoon;Kwak Myounghai
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.299-314
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    • 2011
  • The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.

Analysis of Genetic Diversity in Thirteen Turfgrass Cultivars Cultivated at Golf Courses Using RAPD Markers (RAPD마커를 이용한 국내골프장의 잔디 13 품종의 유전적 다양성 분석)

  • Kim, Min-Jeong;Kim, Tae-Soo;Shim, Chang-Ki;Kim, Yong-Ki;Jee, Hyeong-Jin
    • Weed & Turfgrass Science
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    • v.1 no.4
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    • pp.57-63
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    • 2012
  • This study was carried our to examine the genetic relationship of 13 commercial turfgrass cultivars using Random Amplified Polymorphic DNA to provide genetic informations more efficient golf course management. Analysis of 56 random hexamer primers generated 13 to 54 polymorphic bands among the 13 cultivars with an average of 30.7 bands per primer. The results of cluster analysis based on RAPDs revealed that three major variety groups: Group I - 'Shadow II', 'Aurora Gold', 'Little Bighorn Blue', 'PennA-1', and 'PennA-4'; Group II - 'Midnight II', 'Prosperity', 'Moon light SLT', 'Bright star SLT', and 'Silver dollar'; and Group III - 'Olympic Gold', 'Silver Star', and 'Tar Heel II'. The genetic similarity coefficients among 13 turfgrass cultivars ranged from 0.039 to 1.0 with highest coefficient in Group III. Studies on morphological characters and the effective molecular markers such as sequence characterized amplified regions are further needed to identify relationships and genetic diversities within species and among species.

Identification of new Breeding Lines by Prunus Persica Cultivar-Specific SCAR Primers (SCAR 마커 개발 및 이를 활용한 국내 육성 복숭아 품종 판별)

  • Han, Sang Eun;Cho, Kang-Hee;Nam, Eun Young;Shin, Il-Sheob;Kim, Chung Hee;Kim, Hyun Ran;Kim, Dae-Hyun
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.495-501
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    • 2010
  • Peaches (Prunus persica) are less popular than the fresh fruits, because their flesh gets soft faster. So many breeders focused on their aim to firmness. Other breeders focused on juiciness, flavor and aroma. Breeding requires much labor, time and money. To reduce these requirements, many scientists develop many SSR, CAPS and SCAR makers. New peach varieties bred in our National Institute of Horticultural & Herbal Science (NIHHS) such as, Cheonhong, Suhong and Harhong are yellow flesh cultivars and Yumyeong, Baekmijosaeng, Baekhyang, Jinmi, Soomee, Mihong, Misshong and Yumee are white flesh cultivars. These peach cultivars are planted in orchard of Korea. To assert breeding cultivar patents and prevent patent disputes, we detected cultivar-specific DNA fragment using 235 sets of Operon RAPD primers, analyzed 134 DNA sequences and constructed SCAR primers. To confirm the cultivar-specific SCAR markers, we applied candidate SCAR primers to 30 peach cultivars widely cultivated in Korea. These selected lines are included father and mother lines that were used to develop new varieties in NIHHS. Using fourteen SCAR primer sets, we characterized thirty cultivars selected. The SCAR marker is expected to serve as molecular evidence distinguishing different peach varieties.

Development of SSLP Marker Targeted to P34 Null Gene in Soybean (콩 P34 단백질 결핍 유전자를 이용한 SSLP 마커 개발)

  • Yang, Kiwoung;Ko, Jong-Min;Lee, Young-Hoon;Jeon, Myeong Gi;Jung, Chan-Sik;Baek, In-Youl;Kim, Hyun-Tae;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.502-506
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    • 2010
  • Soybean seed possesses about 15 allergenic proteins recognized by IgEs from soy-sensitive human. The allergenic impact of soybean proteins limit its extensive usage in a broad range of processed foods. Soybean protein P34 or Gly m Bd 30k of the cysteine protease family is one of the major allergen of the soybean seed. P34-null soybean, PI567476, was identified among soybean (Glycine max & Glycine soja Sieb. and Zucc) of approximately 16,226 accessions from USDA soybean germplasm screened. Also, for P34 gene (Williams 82; whole genome sequence cultivar) and P34 null gene (PI567476) comparative analysis of sequences listed in the NCBI database showed the presence of a SSLP (Simple Sequence Length Polymorphism) of 4 base pair. So, a SSLP marker was designed to reveal the polymorphism of the locus. In this study, a population of 339 $F_2$ recombinant inbred lines generated by cross between Taekwang (Glycine max) and PI567476 was used to select $F_{2:3}$ plant of a P34 null gene. The result separation rate Taekwang type, heterozygous type and PI567476 type were shown in 85: 187: 67 since single gene is concerned in as the separation rate of 1:2:1 in $X^2{_{0.05}}=5.99$, df=2. In future, selected plant will identify protein level, whether P34 null protein is equal to P34 null gene.