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      An Improved Method for Cultivar Identification in Radish (Raphanus sativus L.) Using M13-Tailed Primers

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      https://www.riss.kr/link?id=A108917036

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      다국어 초록 (Multilingual Abstract) kakao i 다국어 번역

      Radish is an important root vegetable in the world, and many cultivars have been developed with various molecular marker systems to identify these cultivars. Recently developed markers for radish cultivar identification require only 11 primer pairs, but they still use conventional PCR with different annealing temperatures and time-consuming gel electrophoresis. To improve the genotyping method, we applied touchdown PCR with 11 primers with M13 tails among 105 radish cultivars. Touchdown PCR successfully generated amplicons in all 11 M13-tailed primers with a condition of annealing temperature starting from 55℃, decreased by 1°C and 33 cycles at 53°C. The 11 M13-tailed primers followed by fragment analysis produced 71 amplicons, which produced more amplicons than gel electrophoresis that produced 23 amplicons. Especially, simple sequence repeats produced more amplicons, 12 on average, than the other marker types. The present study requires less effort and provides more accurate results compared to genotyping using gel electrophoresis. Besides, a database can be established using digitized genotyping results among radish cultivars.
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      Radish is an important root vegetable in the world, and many cultivars have been developed with various molecular marker systems to identify these cultivars. Recently developed markers for radish cultivar identification require only 11 primer pairs, b...

      Radish is an important root vegetable in the world, and many cultivars have been developed with various molecular marker systems to identify these cultivars. Recently developed markers for radish cultivar identification require only 11 primer pairs, but they still use conventional PCR with different annealing temperatures and time-consuming gel electrophoresis. To improve the genotyping method, we applied touchdown PCR with 11 primers with M13 tails among 105 radish cultivars. Touchdown PCR successfully generated amplicons in all 11 M13-tailed primers with a condition of annealing temperature starting from 55℃, decreased by 1°C and 33 cycles at 53°C. The 11 M13-tailed primers followed by fragment analysis produced 71 amplicons, which produced more amplicons than gel electrophoresis that produced 23 amplicons. Especially, simple sequence repeats produced more amplicons, 12 on average, than the other marker types. The present study requires less effort and provides more accurate results compared to genotyping using gel electrophoresis. Besides, a database can be established using digitized genotyping results among radish cultivars.

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      참고문헌 (Reference)

      1 최우진 ; 이선애 ; 유수미 ; 강정훈 ; 고호철 ; 이수성, "한국 재래종 무 및 갯무의 RAPD 분석에 의한 유연관계" 한국원예학회 26 (26): 427-431, 2008

      2 정운화 ; 이용범 ; 오종혁 ; 김영규 ; 안춘희 ; 이광식 ; 최수련 ; 임용표 ; 박수형 ; 최기영, "분자표지를 활용한 고품질 가공용 고순도 무 품종 육성" 한국식물생명공학회 41 (41): 56-63, 2014

      3 김호일 ; 홍창표 ; 임수빈 ; 최수련 ; 임용표, "배추의 분자 마커 개발 및 육종적 활용" 한국원예학회 32 (32): 745-752, 2014

      4 Choe, Y., "Studies for similarity evaluation of radish and Chinese cabbage cultivars" 20 : 160-167, 2002

      5 Kang, E., "Reproductive traits and molecular evidence related to the global distribution of cultivated radish (Raphanus sativus L.)" 302 : 1367-1380, 2016

      6 Rohlf, F.J., "NTSYS–pc: Numerical Taxonomy and Multivariate Analysis System Version 2.1" Applied Biostatistics Inc 43-, 2000

      7 Lorenz, E., "M13-tailed primers improve the readability and usability of microsatellite analyses performed with two different allele sizing methods" 31 : 24-26, 2001

      8 Doyle, J. J., "Isolation of plant DNA from fresh tissue" 12 : 13-15, 1990

      9 배경미 ; 심성철 ; 홍지화 ; 최근진 ; 김도훈 ; 권용삼, "Development of Genomic SSR Markers and Genetic Diversity Analysis in Cultivated Radish (Raphanus sativus L.)" 한국원예학회 56 (56): 216-224, 2015

      10 Wang, J. L., "Development and characterization of cDNA library based novel EST-SSR marker in radish (Raphanus sativus L.)" 140 : 164-172, 2012

      1 최우진 ; 이선애 ; 유수미 ; 강정훈 ; 고호철 ; 이수성, "한국 재래종 무 및 갯무의 RAPD 분석에 의한 유연관계" 한국원예학회 26 (26): 427-431, 2008

      2 정운화 ; 이용범 ; 오종혁 ; 김영규 ; 안춘희 ; 이광식 ; 최수련 ; 임용표 ; 박수형 ; 최기영, "분자표지를 활용한 고품질 가공용 고순도 무 품종 육성" 한국식물생명공학회 41 (41): 56-63, 2014

      3 김호일 ; 홍창표 ; 임수빈 ; 최수련 ; 임용표, "배추의 분자 마커 개발 및 육종적 활용" 한국원예학회 32 (32): 745-752, 2014

      4 Choe, Y., "Studies for similarity evaluation of radish and Chinese cabbage cultivars" 20 : 160-167, 2002

      5 Kang, E., "Reproductive traits and molecular evidence related to the global distribution of cultivated radish (Raphanus sativus L.)" 302 : 1367-1380, 2016

      6 Rohlf, F.J., "NTSYS–pc: Numerical Taxonomy and Multivariate Analysis System Version 2.1" Applied Biostatistics Inc 43-, 2000

      7 Lorenz, E., "M13-tailed primers improve the readability and usability of microsatellite analyses performed with two different allele sizing methods" 31 : 24-26, 2001

      8 Doyle, J. J., "Isolation of plant DNA from fresh tissue" 12 : 13-15, 1990

      9 배경미 ; 심성철 ; 홍지화 ; 최근진 ; 김도훈 ; 권용삼, "Development of Genomic SSR Markers and Genetic Diversity Analysis in Cultivated Radish (Raphanus sativus L.)" 한국원예학회 56 (56): 216-224, 2015

      10 Wang, J. L., "Development and characterization of cDNA library based novel EST-SSR marker in radish (Raphanus sativus L.)" 140 : 164-172, 2012

      11 Liu, L., "DNA fingerprinting and genetic diversity analysis of late–bolting radish cultivars with RAPD, ISSR and SRAP markers" 116 : 240-247, 2008

      12 Mun, J., "Construction of a reference genetic map of Raphanus sativus based on genotyping by whole?genome resequencing" 128 : 259-272, 2014

      13 Lee, O., "Assessment of genetic diversity in cultivated radishes (Raphanus sativus) by agronomic traits and SSR markers" 223 : 19-30, 2017

      14 Hong, H., "An economic method to identify cultivars and elite lines in radish (Raphanus sativus L.) for small seed companies and independent breeders" 9 : 140-, 2023

      15 Schuelke, M., "An economic method for the fluorescent labeling of PCR fragments" 18 : 233-234, 2000

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