• Title/Summary/Keyword: 분자마커선발

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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.

Marker-Assisted Selection for Monoecy in Chamoe (Cucumis melo L.) (성발현 연관 분자마커를 이용한 단성화 참외 선발)

  • Bang, Sun-Woong;Song, Kihwan;Sim, Sung Chur;Chung, Sang Min
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.134-141
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    • 2016
  • The DNA marker T1ex, originally developed from melon (Cucumis melo L.) for monoecy, was employed in chamoe, which is referred to as oriental melon. This marker shows size variations in monoecious melon. However, in chamoe, no such detrimental size variation was found in monoecious chamoe, and 99% association between flower phenotypes and genotypes of the T1ex marker was observed in 106 lines of chamoe. To evaluate the efficacy of the T1ex marker for marker-assisted selection (MAS), a total of 240 plants of chamoe breeding lines were screened using the T1ex marker. Among these, 98 varieties were selected. Although the T1ex marker might not be useful for MAS in melon, we found 100% concordance between genotypes and phenotypes for sex expression in chamoe. These results suggest that the T1ex marker will be a useful resource for MAS for monoecy in chamoe.

Cloning of Molecular Marker for Cultivar Protection and Transfer to Nicotiana tabacum L. (품종보호를 위한 분자 마커의 Cloning 및 담배로의 전이)

  • Ku, Ja Jung;Park, Young Doo;Choi, Geun Won
    • Horticultural Science & Technology
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    • v.17 no.6
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    • pp.770-772
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    • 1999
  • This study was conducted to provide a basic system to develop a molecular marker for plant cultivar protection using a recombinant DNA technology. Using Nicotiana tabacum L. plants, the potentiality in the utilization of the developed marker was examined. After homology test with several plant genomes, mouse adenosine deaminase (ADA) gene was selected as DNA source of a molecular marker for cultivar protection. As a result of the digestion of ADA gene with BamHI and Pst I, six DNA fragments were obtained, and 513 bp DNA fragment among them was selected as a possible DNA marker for cultivar protection. Selected 513 bp DNA fragment was efficiently inserted into pBI101 plasmid vector for plant transformation by using phagemid vector pBluescript II SK (+/-) as an intermediate vector. The recombinant pBI101, carrying 513 bp DNA fragment, possible markers for cultivar protection, was transformed into A. tumefaciens LBA4404. Nicotiana tabacum was transformed with A. tumefaciens LBA4404 having the recombinant pBI101 and was confirmed the transfer of 513 bp DNA fragment, a possible molecular marker for cultivar protection.

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Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria×ananassa Duchesne) (딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발)

  • Je, Hee-Jeong;Ahn, Jae-Wook;Yoon, Hae-Suk;Kim, Min-Keun;Ryu, Jae-San;Hong, Kwang-Pyo;Lee, Sang-Dae;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.722-729
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    • 2015
  • Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

Tag-SNP selection and online database construction for haplotype-based marker development in tomato (유전자 단위 haplotype을 대변하는 토마토 Tag-SNP 선발 및 웹 데이터베이스 구축)

  • Jeong, Hye-ri;Lee, Bo-Mi;Lee, Bong-Woo;Oh, Jae-Eun;Lee, Jeong-Hee;Kim, Ji-Eun;Jo, Sung-Hwan
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.218-226
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    • 2020
  • This report describes methods for selecting informative single nucleotide polymorphisms (SNPs), and the development of an online Solanaceae genome database, using 234 tomato resequencing data entries deposited in the NCBI SRA database. The 126 accessions of Solanum lycopersicum, 68 accessions of Solanum lycopersicum var. cerasiforme, and 33 accessions of Solanum pimpinellifolium, which are frequently used for breeding, and some wild-species tomato accessions were included in the analysis. To select tag-SNPs, we identified 29,504,960 SNPs in 234 tomatoes and then separated the SNPs in the genic and intergenic regions according to gene annotation. All tag-SNP were selected from non-synonymous SNPs among the SNPs present in the gene region and, as a result, we obtained tag-SNP from 13,845 genes. When there were no non-synonymous SNPs in the gene, the genes were selected from synonymous SNPs. The total number of tag-SNPs selected was 27,539. To increase the usefulness of the information, a Solanaceae genome database website, TGsol (http://tgsol. seeders.co.kr/), was constructed to allow users to search for detailed information on resources, SNPs, haplotype, and tag-SNPs. The user can search the tag-SNP and flanking sequences for each gene by searching for a gene name or gene position through the genome browser. This website can be used to efficiently search for genes related to traits or to develop molecular markers.

Development of Molecular Markers and Application for Breeding in Chinese Cabbage (배추의 분자 마커 개발 및 육종적 활용)

  • Kim, Ho-Il;Hong, Chang Pyo;Im, Subin;Choi, Su Ryun;Lim, Yong Pyo
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.745-752
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    • 2014
  • Chinese cabbage (Brassica rapa L. ssp. pekinensis) is an economically important vegetable crop as a source of the traditional food Kimchi in Korea. Although many varieties exhibiting desirable traits have been developed by the conventional selective breeding approach, breeding related to abiotic or biotic stresses, such as a particular pests or diseases, or tolerance to climatic conditions, is likely to be slow. This could be helped by an efficient method for selection from various, rapidly-evolved genetic resources on the basis of molecular markers. In particular, the Brassica genome sequencing project enables genome-wide discovery of genes or genetic variants associated with agricultural traits. We here discuss the recent progress in the field of Chinese cabbage breeding with regard to the application of molecular markers.

Genomics and Molecular Markers for Major Cucurbitaceae Crops (주요 박과작물의 유전체 및 분자마커 연구 현황)

  • Park, Girim;Kim, Nahui;Park, Younghoon
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1059-1071
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    • 2015
  • Watermelon and melon are economically important Cucurbitaceae crops. Recently, the development of molecular markers based on the construction of genetic linkage maps and detection of DNA sequence variants through next generation sequencing are essential as molecular breeding strategies for crop improvement that uses marker-assisted selection and backcrossing. In this paper, we intended to provide useful information for molecular breeding of watermelon and melon by analyzing the current status of international and domestic research efforts on genomics and molecular markers. Due to diverse genetic maps constructed and the reference genome sequencing completed in the past, DNA markers that are useful for selecting important traits including yield, fruit quality, and disease resistances have been reported and publicly available. To date, more than 16 genetic maps and loci and linked markers for more than 40 traits have reported for each watermelon and melon. Furthermore, the functional genes that are responsible for those traits are being continuously discovered by high-density genetic map and map-based cloning. In addition, whole genome resequencing of various germplasm is under progress based on the reference genome. Not only by the efforts for developing novel molecular markers, but application of public marker information currently available will greatly facilitate breeding process through genomics-assisted breeding.

Comparison of the complete chloroplast genome sequence of Solanum stoloniferum with other Solanum species generates PCR-based markers specific for Solanum stoloniferum (엽록체 전장유전체 정보를 이용한 감자 야생종 Solanum stoloniferum 구별 분자 마커 개발)

  • Kim, Soojung;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.131-140
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    • 2020
  • Solanum stoloniferum, one of the wild tetraploid Solanum species belonging to the Solanaceae family, is an excellent resource for potato breeding owing to its resistance to several important pathogens. However, the sexual hybridization of S. stoloniferum with S. tuberosum (potato) is hampered due to the sexual incompatibility between the two species. To overcome this and introgress the various novel traits of S. stoloniferum in cultivated potatoes, cell fusion can be performed. The identification of the fusion products is crucial and can be achieved with the aid of molecular markers. In this study, the chloroplast genome sequence of S. stoloniferum was obtained by next-generation sequencing technology, and compared with that of six other Solanum species to identify S. stoloniferum-specific molecular markers. The length of the complete chloroplast genome of S. stoloniferum was found to be 155,567 bp. The structural organization of the chloroplast genome of S. stoloniferum was similar to that of the six other Solanum species studied. Phylogenetic analysis of S. stoloniferum with nine other Solanaceae family members revealed that S. stoloniferum was most closely related to S. berthaultii. Additional comparison of the complete chloroplast genome sequence of S. stoloniferum with that of five Solanum species revealed the presence of six InDels and 39 SNPs specific to S. stoloniferum. Based on these InDels and SNPs, four PCR-based markers were developed to differentiate S. stoloniferum from other Solanum species. These markers will facilitate the selection of fusion products and accelerate potato breeding using S. stoloniferum.

Identification of a Single Nucleotide Polymorphism (SNP) Marker for the Detection of Enhanced Honey Production in Hoenybee (수밀력 우수 꿀벌 계통 판별을 위한 계통 특이 분자마커 개발)

  • Kim, Hye-Kyung;Lee, Myeong-Lyeol;Lee, Man-Young;Choi, Yong-Soo;Kim, Dongwon;Kang, Ah Rang
    • Journal of Apiculture
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    • v.32 no.3
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    • pp.147-154
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    • 2017
  • Honeybees (Apis mellifera) are common pollinators and important insects studied in agriculture, ecology and basic research. Recently, RDA (Rural Development Administration) and YIRI (Yecheon-gun Industrial Insect Research Institute) have been breeding a triple crossbred honey bee named Jangwon, which have the ability to produce superior quality honey. In this study, we identified a single nucleotide polymorphism (SNP) marker in the genome of Jangwon honeybee, particularly, in the paternal line (D line). Initially, we performed Sequence-Based Genotyping (SBG) using the Illumina Hiseq 2500 in 5 honeybee inbred lines; A, C, D, E, and F; and obtained 1,029 SNPs. Seventeen SNPs for each inbred line were generated and selected after further filtering of the SNP dataset. The 17 SNP markers validated by performing TaqMan probe-based real-time PCR and genotyping analysis was conducted. Genotyping analysis of the 5 honeybee inbred lines and one hybrid line, $D{\times}F$, revealed that one set of SNP marker, AmD9, precisely discriminated the inbred line D from the others. Our results suggest that the identified SNP marker, AmD9, is successful in distinguishing the inbred honeybee lines D, and can be directly used for genotyping and breeding applications.

Identification of Pleurotus ostreatus cultivars with the application of multiplex-simple sequence repeat markers (Multiplex SSR마커를 이용한 느타리(Pleurotus ostreatus) 품종 판별)

  • Choi, Jong In;Jung, Hwa Jin;Na, Kyeong sook;Oh, Min-Ji;Kim, Min-Keun;Ryu, Jae-San
    • Journal of Mushroom
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    • v.19 no.1
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    • pp.76-80
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    • 2021
  • To develop a method for the differentiation of Pleurotus ostratus cultivars, the multiplex-simple sequence repeat (SSR) primer set based on the SSRs obtained from whole genomic DNA sequence analysis was designed with two polymerase chain reaction (PCR) primer sets. These SSR primer sets were employed to distinguish 10 cultivars and strains. Twenty polymorphic markers were selected based on the genotyping results. PCR with each primer produced 1-4 distinct bands ranging in size from 150 to 350 bp, which was within the expected range. However, since a sole SSR marker was unable to detect polymorphisms in every cultivar, multiplex PCRs with composite PCR primer sets were employed. The multiplex primer, "166+115," completely discriminated 12 cultivars and strains with 40 loci, which were 12 more than the simple arithmetic addition of each locus of the primers 115 and 166. These results might be useful to provide an efficient method for the differentiation of P. ostreatus cultivars with separate PCRs for the quality control of spawn and protection of breeders' rights.