• 제목/요약/키워드: Primer combinations

검색결과 89건 처리시간 0.023초

Linkage Map Construction and Molecular Genetic Approach in Capsicum spp.

  • Kim, Byung-Dong
    • 식물조직배양학회지
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    • 제27권5호
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    • pp.367-373
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    • 2000
  • We have constructed a molecular linkage map of chili pepper (Capsicum spp.) in an interspecific (C. annuum cv. TF68 x C. chinense cv. Habanero) F$_2$ population of 107 plants with 150 RFLP and 430 AFLP markers. The resulting linkage map consists of 11 large (206-60.3 cM) and 5 small (32.6- 10.3 cM) linkage groups cover-ing 1,320 cM with an average map distance between framework markers of 7.5 cM. Most (80%) of the RFLP markers were pepper-derived clones and these markers were evenly distributed across the genome. By using 30 primer combinations, 444 AFLP markers were generated in the F$_2$population. The majority of the AFLP markers clustered in each linkage group, although PstI/MseI markers were more evenly distributed than Eco RI/MseI markers within the linkage groups. Genes for biosynthesis of carotenoids and capsaicinoids were mapped on our linkage map. This map will provide the basis of studying secondary metabolites in pepper.

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DNA Fingerprinting of Rice Cultivars using AFLP and RAPD Markers

  • Cho, Young-Chan;Shin, Young-Seop;Ahn, Sang-Nag;Gleen B. Gregorio;Kang, Kyong-Ho;Darshan Brar;Moon, Huhn-Pal
    • 한국작물학회지
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    • 제44권1호
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    • pp.26-31
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    • 1999
  • This experiment was conducted to evaluate genetic variation in 48 rice accessions (Oryza sativa L.) using AFLP and RAPD markers. For AFLP, a total of 928 bands were generated with 11 primer combinations and 327 bands (35.2%) of them were polymorphic among 48 accessions. In RAPD analyses using 22 random primers 145 bands were produced, and 121 (83.4%) were polymorphic among 48 accessions. Each accession revealed a distinct fingerprint by two DNA marker systems. Cluster analysis using AFLP-based genetic similarity tended to classify rice cultivars into different groups corresponding to their varietal types and breeding pedigrees, but not using RAPD-based genetic similarity. The AFLP marker system was more sensitive than RAPD in fingerprinting of rice cultivars with narrow genetic diversity.

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AFLP Fingerprinting of Brassica campestis L. ssp. napus var. nippo-oleifera Makino from Korea

  • Huh, Man-Kyu;Huh, Hong-Wook
    • Animal cells and systems
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    • 제5권2호
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    • pp.101-106
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    • 2001
  • AFLPS (amplified fragment length polymorphisms) were used to estimate the genetic diversity of seven populations of Brassica campestis L. ssp. napus var. nippo-oleifera Makino between naturalized and cultivated populations. The seven Korean populations maintained a high level of genetic diversity. For example, all eight primers were high polymorphic, with an average of 3.2 effective alleles per primer set, and the expected heterozygosity was also high. The majority of genetic variance resided within populations The combinations of an insect-pollinated, outcrossing breeding system, large populations sizes, a high degree of gene flow and a propensity for high fecundity may explain the high level of genetic diversity within cultivated populations. Estimates of genetic similarity on the proportion of shared fragments ranged from 0.952 to 0.999. The high level of gene flow In Korean naturalized populations is mainly caused by seed dispersal via sea tide and the gene flow of cultivated populations may be enhanced in part by artificial pollen dispersal.

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Multiplex TaqMan qPCR Assay for Detection, Identification, and Quantification of Three Sclerotinia Species

  • Dong Jae Lee;Jin A Lee;Dae-Han Chae;Hwi-Seo Jang;Young-Joon Choi;Dalsoo Kim
    • Mycobiology
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    • 제50권5호
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    • pp.382-388
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    • 2022
  • White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.

AFLP 마커를 이용한 소규모 사시나무림의 공간적 유전구조 구명 (Fine-scale Spatial Genetic Structure of a Small Natural Stand of Populus davidiana in South Korea using AFLP markers)

  • 이민우;홍경낙;박유진;이제완;임효인
    • 한국산림과학회지
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    • 제105권3호
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    • pp.309-314
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    • 2016
  • 변화하는 자연환경에서 식물이 생존하기 위해서는 적절한 유전다양성을 유지할 뿐 아니라 지역적응성을 갖추어야 세대를 성공적으로 이어나갈 수 있다. 만약 유전다양성이 급격히 감소하게 된다면 집단이 쇠퇴하고 소멸 위험성이 커지게 된다. 본 연구는 주변 집단으로 부터 화분이나 종자의 유입이 어려운 소규모 사시나무 집단의 유전구조를 구명하였다. 월악산 미륵리의 사시나무림은 전체 분포면적 $14,000m^2$에 성목은 350개체로 추정되며, 임분내에 설정한 $70m{\times}70m$ 조사구에 출현하는 123개체 중 61개체를 대상으로 AFLP 마커를 이용하여 유전변이를 분석하였다. 조사구내 사시나무의 수령은 평균 16년 최고 32년생이었으며, 개체의 공간적 분포는 약한 밀집 형태를 이루고 있었다. AFLP primer 6조합에서 196개 증폭산물을 확인하였으며, 이 중 151개는 다형성을 보였다. primer 조합당 평균 유전자좌수는 32.7(표준편차=7.2), 이형접합도 기대치($H_e$)는 0.154, Shannon의 다양성 지수(S.I.)는 0.254로 나타나서, 월악산 사시나무는 우리나라 사시나무 집단 평균에 비하여 매우 낮은 유전다양성을 갖고 있는 것으로 나타났다. 공간적 유전구조는 24 m 이내에서 분포하는 개체들 간에 유전적 유사성이 나타났으며, 소규모 면적과 고립된 분포지 특성으로 인하여 비교적 작은 유전군락이 형성된 것으로 생각된다.

Association between Single Nucleotide Polymorphisms of Fatty Acid Synthase and Fat Deposition in the Liver of the Overfed Goose

  • Wu, Wei;Guo, Xuan;Zhang, Lei;Hu, Dan
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권9호
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    • pp.1244-1249
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    • 2014
  • Goose fatty liver is one of the most delicious and popular foods in the world, but there is no reliable genetic marker for the early selection and breeding of geese with good liver-producing potential. In our study, one hundred and twenty-four 78-day-old Landes geese bred in Shunda Landes goose breeding farm, Jiutai, Jilin, China were selected randomly. The fatty livers were sampled each week after overfeeding during a three week period. Polymerase chain reaction-single strand conformation polymorphism and DNA sequencing were used to identify single nucleotide polymorphisms (SNPs) of fatty acid synthase (FAS), which is an important enzyme involved in the synthesis of fat under both physiological and pathological conditions. Least-squares correlation was established between these SNPs and fatty liver weight, abdominal fat weight, and intestinal fat weight of the overfed Landes geese, respectively. The results showed that fatty liver weight of geese with EF and FF genotypes (amplified by primer P1) was significantly higher than that of the EE genotype (p<0.05), and liver weight of CD and DD genotypes (amplified by primer P2) was significantly higher than that of the CC genotype (p<0.05). Different genotype combinations showed different liver weights, and from highest to lowest were ABDD, DDEF, DDFF, DDEE, ABEF, ABFF, AADD, and CDEF. Further analysis of DNA sequencing showed that there were two SNPs within the 5' promoter region the FAS gene. The geese of EF and FF genotypes carried a change of T to C, and the geese of CD and DD genotypes carried a change of A to G. The changes of the bases could potentially influence the binding of some transcription factors to this region as to regulate FAS gene. To our knowledge, this is the first report of SNPs found within the 5' promoter region of the Landes goose FAS gene, and our data will provide an insight for early selection of geese for liver production.

Genetic diversity, structure analysis and relationship in soybean mutants as revealed by TRAP marker

  • Kim, Dong-Gun;Lyu, Jae-Il;Lee, Min-Kyu;Kim, Jung Min;Hong, Min Jeong;Kim, Jin-Baek;Bae, Chang-Hyu;Kwon, Soon-Jae
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 추계학술대회
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    • pp.43-43
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    • 2018
  • Mutation breeding by radiation is useful for improving various crop species. Up to now, a total of 170 soybean mutant varieties have been released in the world, which is the second most registered varieties after rice. Despite the economic importance of soybean, there have been no TRAP marker system studies on genetic relationships between/among mutant lines. To develop a strategy of Mutant Diversity Pool (MDP) conservation, a study on the genetic diversity of 210 soybean mutant lines (8 cultivars and 202 mutants) was performed through a TRAP analysis. Sixteen primer combinations amplified a total of 551 fragments. The highest (84.00%) and lowest (32.35%) polymorphism levels were obtained with primers MIR157B + Ga5 and B14G14B + Ga3, respectively. The mean PIC values 0.15 varied among the primer combination ranging from 0.07 in B14G14B + Sal2 to 0.23 in MIR157B + Sa4. Phylogenetic, principal component analysis (PCA) and structure analysis indicated that the 210 lines belong to four groups based on the 16 combination TRAP markers. AMOVA showed 21.0% and 79.0% variations among and within the population, respectively. Overall, the genetic similarity of each cultivar and its mutants were higher than within other mutant populations. Our results suggest that the TRAP marker system may be useful for assessing the genetic diversity among soybean mutants and help to improve our knowledge of soybean mutation breeding.

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우리나라 주요 강과 호수에 분포하는 외래어종 배스 Micropterus salmoides의 AFLP 분석에 의한 유전적 분화 (Genetic Differentiation of the Largemouth Bass Micropterus salmoides from the Major Rivers and Reservoirs in Korea Assessed by AFLP)

  • 이완옥;이일로;송하윤;방인철
    • 생태와환경
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    • 제41권3호
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    • pp.395-401
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    • 2008
  • 한국에 도입된 외래어종 배스 Micropterus salmoides 9 집단의 유전 다양성 및 집단구조를 AFLP 분석을 통해 조사하였다. 3쌍의 primer 조합을 이용한 AFLP 분석에서 총 299.2개의 밴드가 생성되어 다형성 밴드의 비율은 14.1$\sim$21%로 유사하게 나타났으며, 이형접합률 (0.054$\sim$0.067) 및 유전적 다양성 (0.069$\sim$0.085)은 낮은 값을 보였다. 집단간 유전적 거리 및 상동성 분석 역시 유사한 결과를 나타내어 본 연구에 사용된 배스 9집단은 유전적으로 매우 밀접한 근연관계를 나타내었다. 또한 집단간 분화도가 비교적 높게 나타나 9집단의 유전적 분화가 빠르게 진행 중인 것으로 나타났다. 따라서 배스 집단의 낮은 유전 다양성과 집단간 분화도 값은 매우 작은 집단이 팔당호에 처음 도입되어 우리나라 전 수계에 확산되었음을 암시한다.

박과 작물 종자전염 바이러스 3종(CGMMV, ZGMMV, KGMMV)의 간편한 동시진단 VC/RT-PCR 유전자 진단 (Triplex Virion Capture (VC)/RT-PCR for Three Seed Transmissible Tobamoviruses of CGMMV, ZGMMV and KGMMV Occurring on Cucurbitaceae)

  • 조점덕;김정수;이신호;정봉남
    • 식물병연구
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    • 제13권2호
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    • pp.82-87
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    • 2007
  • 박과 작물에 발생하는 종자전염성 막대형 바이러스인 CGMMV, KGMMV, ZGMMV 3종의 단독 및 동시 VC/RT-PCR 유전자 진단법을 개발하였다. CGMMV 등 3종의 바이러스에 대한 동시진단용 조합선발을 위하여 12조합 중에서 동시 진단용 프라이머 조합 CGMMV-C724, KGMMV-K513, ZGMMV-Z407A를 선발하였다. CGMMV등 3종에 대한 triplex VC/RT-PCR 유전자 진단법은 박, 수박, 오이, 멜론, 쥬키니 호박, 애호박, 담배(N. benthamiana) 작물의 식물체 즙액과 프라이머 간에 간섭현상 없이 특이적으로 진단되었다.

Genetic Diversity of Sweet potato feathery mottle virus from Sweet Potatoes in Korea

  • Kwak, Hae-Ryun;Kim, Mi-Kyeong;Jung, Mi-Nam;Lee, Su-Heon;Park, Jin-Woo;Kim, Kook-Hyung;Ko, Sug-Ju;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • 제23권1호
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    • pp.13-21
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    • 2007
  • Sweet potato feathery mottle virus(SPFMV) is one of the most prevalent viruses infecting sweet potatoes and occurs widely in sweet potato cultivating areas in Korea. To assess their genetic variation, a total of 28 samples infected with SPFMV were subjected to restriction fragment length polymorphism(RFLP) analysis using DNAs amplified by RT-PCR with specific primer sets corresponding to the coat protein(CP) region of the virus. The similarity matrix by UPGMA procedure indicated that 28 samples infected with SPFMV were classified into three groups based on the number and size of DNA fragments by digestion of CP-encoding regions with 7 enzymes including SalI, AluI, EcoRI, HindIII, FokI, Sau3AI, and DraI bands. Four primer combinations out of 5 designed sets were able to differentiate SPFMV and sweet potato virus G infection, suggesting that these specific primers could be used to differentiate inter-groups of SPFMV. Sequence analysis of the CP genes of 17 SPFMV samples were 97-99% and 91-93% identical at the intra-group and inter-groups of SPFMV, respectively. The N-terminal region of the CP is highly variable and examination of the multiple alignments of amino acid sequences revealed two residues(residues 31 and 32) that were consistently different between SPFMV-O and SPFMV-RC.