• Title/Summary/Keyword: Cleaved Amplified Polymorphic Sequence

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Marker Development for Erect versus Pendant-Orientated Fruit in Capsicum annuum L.

  • Lee, Heung-Ryul;Cho, Myeong-Cheoul;Kim, Hyoun-Joung;Park, Sung-Woo;Kim, Byung-Dong
    • Molecules and Cells
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    • v.26 no.6
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    • pp.548-553
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    • 2008
  • The erect habit of fruit setting is a unique characteristic of ornamental peppers and wild pepper species. The erect habit is known to be controlled by the up locus on pepper (Capsicum annuum L.) chromosome 12. The result of a genetic analysis using Saengryeog 211 (pendant), Saengryeog 213 (erect), and their $F_1$ and $BC_1$ progeny demonstrated that up is a recessive gene. To develop an up-linked marker, bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) were employed using 108 $F_{2:3}$ individuals. The closest AFLP marker, $A2C7_{469}$, was located at a genetic distance of 1.7 cM from the up locus and was converted into a cleaved amplified polymorphic sequence (CAPS) marker. This marker was mapped at a genetic distance of 4.3 cM from the up locus. When the CAPS was applied to seven ornamental lines and 27 breeding lines with erect fruit, these genotypes of 28 lines were correctly predicted. Thus, the CAPS marker will be useful for marker-assisted selection (MAS) of pepper breeding lines with the up allele at the early seedling stage.

Development of a CAPS marker for the identification of the Lentinula edodes cultivar, 'Sanmaru 2ho' (표고버섯 품종 '산마루2호'를 구분할 수 있는 CAPS marker 개발)

  • Moon, SuYun;Lee, Hwa-Yong;Ka, Kang-Hyeon;Koo, Chang-Duck;Ryu, HoJin
    • Journal of Mushroom
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    • v.16 no.1
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    • pp.51-56
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    • 2018
  • In Korea, the oak mushroom (Lentinula edodes) is highly preferred by consumers in the food industry and makes up about 97.7% of the total forest mushroom production. This indicates that the oak mushroom is an important non-timber forest product in Korea. Recently, the breeding and development of new cultivars of L. edodes have been actively initiated, and the development of molecular markers that are able to identify and discriminate the new cultivars is crucial for protecting the breeder's rights. This study was carried out to develop a cleaved amplified polymorphic sequence (CAPS) marker for the identification and discrimination of a new cultivar, Sanmaru 2ho from the 37 other oak mushroom cultivars. A single nucleotide polymorphism (SNP) was identified at the $1,803,483^{rd}$ position of scaffold2 in the genome of Sanmaru 2ho. The amplified DNA containing the SNP of Sanmaru 2ho was uniquely not cleaved by the restriction enzyme, Hha I, and thus Sanmaru 2ho was successfully distinguished from the other oak mushroom cultivars.

Discrimination of Korean Agaricus bisporus cultivars using CAPS markers (CAPS 마커를 이용한 국내 개발 양송이 품종 구분)

  • Lee, Hwa-Yong;An, Hyejin;Oh, Youn-Lee;Jang, Kab-Yeul;Chung, Jong-Wook
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.336-340
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    • 2021
  • The cleaved amplified polymorphic sequence (CAPS) marker uses a restriction enzyme recognition site resulting from single nucleotide polymorphisms and insertions and deletions on the DNA sequence. This technique does not require expensive equipment, the process is simple, and clear results can be obtained reliably. In this study, Agaricus bisporus cultivars SaeA, SaeDo, SaeHan, SaeYeon, SaeJeong, Dodam, Seolgang, Dahyang, Hogam, and Hadam developed in Korea were discriminated using four CAPS markers. Our results indicated that it is possible to distinguish the ten cultivars and determine the genetic diversity among them.

Identification of Korean Strawberry Cultivars using DNA markers (DNA 표지를 이용한 딸기 국내 육성 품종 판별)

  • Cho, Kang-Hee;Rho, Il Rae;Cho, Yong Seop;Park, Pue-Hee
    • Korean Journal of Breeding Science
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    • v.40 no.4
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    • pp.401-407
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    • 2008
  • This study was conducted to develop the DNA markers for identification of the strawberry cultivars in Korea and Japan. We developed fifteen cleaved amplified polymorphic sequence (CAPS) markers based on the Fragaria gene sequences. Among them six CAPS markers showed polymorphism exclusively in one cultivar. Five CAPS markers (ANR-MspI, ANR-BamHI, ACO-HinfI, DFR-AseI, FGT-MspI) provided enough polymorphism to identify eight Korean strawberry cultivars except for 'Maehyang' and 'Sunhong'. To complement the fifteen CAPS markers, we selected another fifteen sequence-related amplified polymorphism (SRAP) and one of them, me1/em5_460bp marker, made it possible to discriminate between 'Maehyang' and 'Sunhong'. Therefore, application of the five CAPS markers and one SRAP marker were sufficient to identify the nineteen Korean and Japanese strawberry cultivars. These markers could be used practically for cultivar identification of Korean and Japanese strawberry.

Development of a Segregating Population with Biological Efficiency in Agaricus bisporus

  • Oh, Youn-Lee;Sonnenberg, Anton S.M.;Baars, Johan J.P.;Jang, Kab-Yeul;Oh, Min ji;Im, Ji-Hoon;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.328-335
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    • 2017
  • In this study, we made a population with high biological efficiency (BE) to investigate the complex genetic architecture of yield-related traits in Agaricus bisporus. MB-013 crossed between bisp 015-p2 and bisp 034-p2, had high BE. Additionally MB-013 was an intervarietal hybrid that intercrosses with A. bisporus var. burnettii, bisp 015, and A. bisporus var. bisporus, bisp 034. One hundred and seventy homokaryons were selected using the cleaved amplified polymorphic sequence (CAPS) markers (PIN primer/HaeIII) from 300 single spore isolates (SSIs). One hundred $BC_1F_1$ hybrids were obtained by crossing the homokaryons of MB-013 with bisp15-p1. The population of 100 BC1F1 hybrids is suitable for analyses of BE.

Development of CAPS marker for identifying a Formosan lily (Lilium formosanum) (흰나리(Lilium formosanum Wallace) 식별을 위한 CAPS 마커의 개발)

  • Chung, Sung Jin;Lee, Ka Youn;Yoon, A Ra;Jang, Ji Young;Kim, Jin Kug;Lee, Geung-Joo
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.101-106
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    • 2014
  • This study was conducted to identify lily species native to Korea from formosan lily (Lilium formosanum) belonging to Longiflorum section. Due to flowering time, flower color and orientation, long shelf life and resistant to diseases, the native lily species can be valuable genetic resources for interspecific hybrids. One of the chloroplast genes, matK, was used to clone and sequence to explore any base changes. The matK was successfully amplified into 1,539 bp (94% of the gene) and phylogenetic tree demonstrated 6 clades for those 11 lily species used in this study. There were one or two base substitutions among 10 lilies native to Korea, while formosan lily native to Taiwan exhibited 6 base substitutions in matK gene, rendering it genetically distant. A restriction enzyme NruI recognized one of the six base changes, and digested the matK gene of 10 native lily species only, but not in formosan lily. The confirmed cleavage characteristic of the target region in matK gene was designed into a CAPS (cleaved amplified polymorphic sequences) marker which will be available to estimate compatibility of interspecific hybridization and to trace the pedigree when those native lilies are crossed with the formosan lily.

Molecular Identification of Zoysia japonica and Zoysia sinica (Zoysia Species) Based on ITS Sequence Analyses and CAPS (ITS 염기서열 분석 및 CAPS를 이용한 조이시아 속(Zoysia) 들잔디와 갯잔디의 구별)

  • Hong, Min-Ji;Yang, Dae-Hwa;Jeong, Ok-Cheol;Kim, Yang-Ji;Park, Mi-Young;Kang, Hong-Gyu;Sun, Hyeon-Jin;Kwon, Yong-Ik;Park, Shin-Young;Yang, Paul;Song, Pill-Soon;Ko, Suk-Min;Lee, Hyo-Yeon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.344-360
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    • 2017
  • Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

Discrimination of Korean Ginseng Cultivars by Sequence Tagged Sites (STS) Markers (STS 마커를 이용한 고려인삼 품종 및 육성계통 판별)

  • Jo, Ick Hyun;Shin, Mi Ran;Kim, Young Chang;Lee, Seung Ho;Kim, Jang Uk;Moon, Ji Young;Noh, Bong Soo;Kang, Sung Taek;Lee, Dong Jin;Hyun, Dong Yun;Kim, Dong Hwi;Kim, Kee Hong;Bang, Kyong Hwan
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.5
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    • pp.353-360
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    • 2013
  • Korean ginseng (P. ginseng C. A. Meyer) is one of the most important medicinal plant in the world. Understanding genetic variability among the assortment of Korean ginseng is important for breeding. The aim of this study was to molecularly characterize Korean ginseng cultivar and breeding lines through the use of eight previously reported STS markers (MFGp183, MFGp130, MFGp110, UFGp74, UFGp163, MFGp108, MFGp81 and UFGp156). All STS markers produced interpretable electropherograms from 31 accessions consisting of 11 Korean ginseng cultivars and 20 breeding lines. When eight STS markers were combined, we identified to total 19 genetic patterns; in particular, nine cultivars (Chunpoong, Yunpoong, Gopoong, Gumpoong, Sunpoong, Sunone, Cheongseon, Sunhyang, Cheonryang) and 5 breeding lines (G08012, G04079, G04075, G08036, G04110) in ginseng samples can be discriminated from the others. Together with other available markers, these STS markers will contribute to the management of ginseng genetic resources and the protection of breeders' rights.

Characterization of the Lsi1 Homologs in Cucurbita moschata and C. ficifolia for Breeding of Stock Cultivars Used for Bloomless Cucumber Production

  • Jung, Jaemin;Kim, Joonyup;Jin, Bingkui;Choi, Youngmi;Hong, Chang Oh;Lee, Hyun Ho;Choi, Youngwhan;Kang, Jumsoon;Park, Younghoon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.333-343
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    • 2017
  • Bloomless cucumber fruits are commercially produced by grafting onto the pumpkin stocks (Cucurbita moschata) to restricted silicon ($SiO_2$) absorption. Inhibition of silicon absorption in bloomless stocks is conferred by a mutant allele of the CmLsi1 homologous to Lsi1 in rice. In this study, we characterized the Lsi1 homologs in pumpkin (C. moschata) and its cold-tolerant wild relative C. ficifolia ('Heukjong') in order to develop a DNA marker for selecting a bloomless trait and to establish the molecular basis for breeding bloomless stock cultivars of C. ficifolia. A Cleaved amplified polymorphic sequence (CAPS) marker (CM1-CAPS) was designed based on a non-sysnonymous single nucleotide polymorphism (SNP, C>T) of the CmLsi1 mutant-type allele, and its applicability for Marker-assisted selection (MAS) was confirmed by evaluating three bloom and five bloomless pumpkin stock cultivars. Quantitative RT-PCR of the CmLsi1 for these stock cultivers implied that expression level of the CmLsi1 gene does not appear to be associated with the bloom/bloomless trait and may differ depending on plant species and tissues. A full length cDNA of the Lsi1 homolog [named CfLsi1($B^+$)] of 'Heukjong' (C. ficifolia), was cloned and sequence comparison between CmLsi1($B^+$) and CfLsi1($B^+$) revealed that there exists total 24 SNPs, of which three were non-synonymous. Phylogenetic analysis of CfLsi1($B^+$) and Lsi1 homologs further revealed that CfLsi1($B^+$) is closesly related to Nodulin 26-like intrinsic proteins (NIPs) and most similar to CpNIP1 of C. pepo than C. moschata.

Improvement of Seedling Establishment in Wet Direct Seeding of Rice using the Anaerobic Germination Tolerance Gene Derived from Weedy Photoblastic Rice (잡초벼 PBR 혐기발아 내성 유전자 활용 벼 담수직파 초기 입모 개선)

  • Jeong, Jong-Min;Mo, Youngjun;Baek, Man-Kee;Kim, Woo-Jae;Cho, Young-Chan;Ha, Su-Kyung;Kim, Jinhee;Jeung, Ji-Ung;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.161-171
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    • 2020
  • Direct seeding is one of the rice seedling establishment methods that is increasingly being practiced by farmers to save labor and reduce costs. However, this method often causes poor germination under flooding conditions after sowing. In this study, we developed japonica elite lines with quantitative trait loci (QTL) associated with anaerobic germination (AG) tolerance to overcome poor germination and seedling establishment in wet direct seeding. The QTL introgression lines were developed from a cross between weedy photoblastic rice as the AG donor and the Nampyeong variety via phenotypic and genotypic selection. Compared to Nampyeong, the survival rates of the selected lines were improved by approximately 50% and 240% under field and greenhouse conditions, respectively. To improve selection efficiency by marker assisted selection, the QTL markers associated with AG tolerance were converted to cleaved amplified polymorphic sequence markers designed based on next-generation sequence analysis. These lines retained similar agronomic traits and yield potential to the parent, Nampyeong. Among these lines, we selected the most promising line, which exhibited high survival rate and good agricultural traits under flooding conditions and named the line as Jeonju643. This line will contribute to breeding programs aiming to develop rice cultivars adapted to wet direct seeding. This study demonstrates the successful application of marker-assisted selection to targeted introgression of anaerobic genes into a premium quality japonica rice variety.