• 제목/요약/키워드: Primer combinations

검색결과 89건 처리시간 0.026초

Construction of Genetic Linkage Map for Korean Soybean Genotypes using Molecular Markers

  • 조예진;박대진;한성진;오주호;황정규;고미숙;정종일
    • 한국작물학회지
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    • 제48권4호
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    • pp.297-302
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    • 2003
  • Genetic linkage maps serve the plant geneticist in a number of ways, from marker assisted selection in plant improvement to map-based cloning in molecular genetic research. Genetic map based upon DNA polymorphism is a powerful tool for the study of qualitative and quantitative traits in crops. The objective of this study was to develop genetic linkage map of soybean using the population derived from the cross of Korean soybean cultivar 'Kwangkyo, and wild accession 'IT182305'. Total 1,000 Operon random primers for RAPD marker, 49 combinations of primer for AFLP marker, and 100 Satt primers for SSR marker were used to screen parental polymorphism. Total 341 markers (242 RAPD, 83 AFLP, and 16 SSR markers) was segregated in 85 $\textrm{F}_2$ population. Forty two markers that shown significantly distorted segregation ratio (1:2:1 for codominant or 3:1 for domimant marker) were not used in mapping procedure. A linkage map was constructed by applying the computer program MAPMAKER/EXP 3.0 to the 299 marker data with LOD 4.0 and maximum distance 50 cM. 176 markers were found to be genetically linked and formed 25 linkage groups. Linkage map spanned 2,292.7 cM across all 25 linkage groups. The average linkage distance between pair of markers among all linkage groups was 13.0 cM. The number of markers per linkage group ranged from 2 to 55. The longest linkage group 3 spanned 967.4 cM with 55 makers. This map requires further saturation with more markers and agronomically important traits will be joined over it.

하나의 나무에서 채집된 자작나무버섯의 균주간 유전적 동일성 확인 (Confirmation of Genetic Homogenecity among Wild Strains of Piptoporus betulinus Collected from a Single Tree)

  • 가강현;장지연;류성렬;윤갑희;박원철
    • 한국균학회지
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    • 제36권1호
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    • pp.63-65
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    • 2008
  • 자작나무버섯의 4개 균주는 설악산의 사스래나무 한 나무에서 분리하였다. 균주들은 균주간의 동일성 유무를 대치배양과 ITS 영역의 일부를 증폭하는 NSI1과 ITS4B primer pairs를 이용하여 염기서열을 분석하였다. 균주들간의 대치선은 PDA 배지상에서 나타나지 않았고, rDNA 염기서열도 동일하게 나타났다. 결론적으로 이들 균주들은 동일한 균으로부터 기원한 것으로 판단되었다.

Several factors affecting on seed germination of Dracocephalum argunense Fischer ex Link

  • Chang, Young-Deug;Lee, Cheol-Hee;Song, Jeong-Seob;Hwang, Ju-Kwang
    • 한국자원식물학회지
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    • 제22권3호
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    • pp.236-241
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    • 2009
  • Dracocephalum argunense Fischer ex Link (Labiatae) is a perennial herbaceous plant used as valuable materials for ornamentals, honey production, and pharmaceutics. Since seed germination of this species was quite difficult, present studies were conducted to improve the germination rate by subjecting the seeds to various environmental conditions (temperature and light) and treatments (scarification, priming and seed coating). Optimum temperature for adequate germination was 20$^{\circ}C$ though it ranged from 15$^{\circ}C$ to 25$^{\circ}C$, and low temperature treatment improved germination rate. Light was required for higher germination rate in this species. The scarification of seeds resulted in much higher germination, especially by the physical treatment with sandpaper or chemical treatment with sulfuric acid for 30 seconds. Various primers with different concentrations were treated on the seeds and it was demonstrated that low temperature enhanced germination rate, regardless of kinds and concentrations of the primers. Three treatment combinations of the primers, 0.5 mM $GA_{3}$ treated for 48 hours, 0.5 mM IAA for 24 hours, and 1.0 mM IAA for 24 hours, increased the seed germination rate profoundly. Soaking treatment of inorganic salts, $KNO_{3}$ and $KH_{3}PO_{4}$, promoted germination when seeds were subjected to low temperature. Water soluble primers such as sucrose at 0.5 and 3% concentration and solid primer talc powder were effective in enhancing germination rate.

Development of Molecular Markers for Xanthomonas axonopodis Resistance in Soybean

  • Kim Ki-Seung;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • 한국작물학회지
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    • 제49권5호
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    • pp.429-433
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    • 2004
  • A single recessive gene, rxp, controls the bacterial leaf pustule (BLP) resistance in soybean and in our previous article, it has been mapped on linkage group (LG) D2 of molecular genetic map of soybean. A total of 130 recombinant inbred lines (RILs) from a cross between BLP-resistant SS2-2 and BLP-susceptible Jangyeobkong were used to identify molecular markers linked to rxp. Fifteen simple sequence repeat (SSR) markers on LG D2 were screened to construct a genetic map of rxp locus. Only four SSR markers, Satt135, Satt372, Satt448, and Satt486, showed parental polymorphisms. Using these markers, genetic scaffold map was constructed covering 26.2cM. Based on the single analysis of variance, Satt372 among these four SSR markers was the most significantly associated with the resistance to BLP. To develop new amplified fragment length polymorphism (AFLP) marker linked to the resistance gene, bulked segregant analysis (BSA) was employed. Resistance and susceptible bulks were made by pooling equal amount of genomic DNAs from ten of each in the segregating population. A total of 192 primer combinations were used to identify specific bands to the resistance, selecting three putative AFLP markers. These AFLP markers produced the fragment present in SS2-2 and the resistant bulk, and not in Jangyeobkong and the susceptible bulk. Linkage analysis revealed that McctEact97 $(P=0.0004,\;R^2=14.67\%)$ was more significant than Satt372, previously reported as the most closely linked marker.

Identification and characterization of novel single nucleotide polymorphism markers for fat deposition in muscle tissue of pigs using amplified fragment length polymorphism

  • Supakankul, Pantaporn;Kumchoo, Tanavadee;Mekchay, Supamit
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권3호
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    • pp.338-346
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    • 2017
  • Objective: This study was conducted to identify and evaluate the effective single nucleotide polymorphism (SNP) markers for fat deposition in the longissimus dorsi muscles of pigs using the amplified fragment length polymorphism (AFLP) approach. Methods: Sixty-four selective primer combinations were used to identify the AFLP markers in the 20 highest- and 20 lowest-intramuscular fat (IMF) content phenotypes. Five AFLP fragments were converted into simple codominant SNP markers. These SNP markers were tested in terms of their association with IMF content and fatty acid (FA) composition traits in 620 commercially crossbred pigs. Results: The SSC7 g.4937240C>G marker showed an association with IMF content (p<0.05). The SSC9 g.5496647_5496662insdel marker showed a significant association with IMF content and arachidonic levels (p<0.05). The SSC10 g.71225134G>A marker revealed an association with palmitoleic and ${\omega}9$ FA levels (p<0.05), while the SSC17 g.61976696G>T marker showed a significant association with IMF content and FA levels of palmitoleic, eicosenoic, arachidonic, monounsaturated fatty acids, and ${\omega}9$ FA levels. However, no significant association of SSC8 g.47338181G>A was observed with any IMF and FA levels in this study. Conclusion: Four SNP markers (SSC7 g.4937240C>G, SSC9 g.5496647_5496662insdel, SSC10 g.71225134G>A, and SSC17 g.61976696G>T) were found to be associated with IMF and/or FA content traits in commercially crossbred pigs. These findings provide evidence of the novel SNP markers as being potentially useful for selecting pigs with the desirable IMF content and FA composition.

저염 발효오이로부터 16S rDNA-PCR과 RFLP분석을 통한 부패균의 신속한 확인 (16S rDNA-PCR and RFLP Analysis for rapid identification of Spoilage Bacteria from low Salt Cucumber Brine)

  • 김재호;장혜영
    • KSBB Journal
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    • 제19권1호
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    • pp.72-77
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    • 2004
  • 건강에 대한 관심의 증대로 인한 저염식품의 개발 필요성에 맞추어 오이발효에 염의 농도를 낮추게 되면 정상적 일차발효 후에 이차발효가 진행됨으로써 결국 부패하게 된다. 장기간에 걸쳐 다양한 균의 복합적 작용으로 진행되는 저염 발효오이의 부패 과정을 이해하기 위하여 이에 관련된 균을 분리 동정하였다. 부패발효액을 무기배양하여 균을 분리하고 이들의 16s rRNA 유전자 부분을 universal primer를 이용한 PCR로서 증폭하여 서열분석 하였다. 분석된 800 염기 길이의 서열 전체를 그대로 이용하여 NCBI의 BLAST로서 유사종을 찾고 RDP의 Sequence Aligner와 Sequence Match에서 재확인하여 분리된 균을 3속 8종으로 동정하였다. Database 내의 표준균 서열을 기반으로 한 제한효소 지도와 동정된 균의 PCR 생성묵의 제한효소 처리결과(RFLP)를 비교하여 동정 결과를 실험으로 검정하였다. 동정과정에서 sequencing 결과 전체를 이용하는 점과 RDP를 통한 확인과 RFLP를 이용한 검정은 동정 결과에 대한 신뢰도를 한층 증가시켰다. 또 분리된 8종은 개별적 특징의 조사나 적절한 조합을 이룰 때의 상호 의존도 등을 조사할 수 있게 함으로써 여러 균에 의한 복합적 과정인 부패를 순차적 혹은 요인별로 나누어 살펴보는 연구를 가능하게 한다.

Genetic Diversity and Population Genetic Structure of Cephalotaxus koreana in South Korea

  • Hong, Kyung Nak;Kim, Young Mi;Park, Yu Jin;Lee, Jei Wan
    • 한국자원식물학회지
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    • 제27권6호
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    • pp.660-670
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    • 2014
  • The Korean plum yew (Cephalotaxus koreana Nakai) is a shade-tolerant, coniferous shrub. The seeds have been used as a folk medicine in Korea, and an alkaloid extract (HTT) is known to have anticancer properties. We estimated the genetic diversity of 429 trees in 16 populations in South Korea using 194 polymorphic amplicons from seven combinations of AFLP primer-restriction enzymes. The average number of effective alleles and the percentage of polymorphic loci were 1.37 and 79.4%, respectively. Shannon's diversity index and the expected heterozygosity were 0.344 and 0.244, respectively. We divided 16 populations into four groups on the UPGMA dendrogram and the PCA biplot. The first two principal components explained 84% of the total genetic variation. Genetic differentiation between populations explained 14% of total genetic variation, and the remaining 86% came from difference between individuals within populations, as determined by an analysis of molecular variance (AMOVA). However, the genetic differentiation did not correlate with the geographic distance between populations from the Mantel test. The Bayesian statistics, which are comparable to Wright's $F_{ST}$ and Nei's $G_{ST}$, were ${\theta}^I=0.406$ and ${\theta}^{II}=0.172$, respectively. The population genetic diversity was slightly lower, and the strength of genetic differentiation was much weaker, than the average of those plants having similar life histories, as assessed using arbitrary marker systems. We discuss strategies for the genetic conservation of the plum yew in Korea.

Variation in trn-L/trn-V and trn-F/trn-T spacer regions of cpDNA in Abies koreana Wilson and A. nephrolepis Traut./Maxim

  • Kormutak, A.;Hong, Y.-P.;Kwon, H.-Y.;Kim, C.-S.
    • 한국산림과학회지
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    • 제96권2호
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    • pp.131-137
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    • 2007
  • The first evidence has been provided about the variation within trnL-trnV and trnF-trnT spacer regions of cpDNAs in Korean fir and Manchurian fir, revealed by PCR-RFLP analysis. Four cpDNA haplotypes have accordingly been recognized by being analyzed using the trnL-trnV/Tru11 primer-enzyme combination and 3 haplotypes using the trnF-trnT/TagI combination, which exhibited inter and intraspecific variation. A total of 6 cpDNA haplotypes were recognized by pooling the PCR-RFLP variants observed in both combinations. Haplotypes 2 and 3 were common for both species investigated, whereas haplotypes 1, 4, and 5 were detected only in Korean fir and haplotype 6 was detected only in Manchurian fir. Although haplotypes 2 and 3 were common in both species, haplotype 2 was major haplotype for Korean fir and haplotype 3 was one of the 2 major haplotypes for Manchurian fir. Restricted occurrence of haplotype 4 in Mt. Halla and haplotype 5 in Mt. Jiri of the Korean fir may represent the existence of geographic isolation by the sea between them. Diagnostic potential of individual haplotypes in discriminating between the two species as well as between their populations is discussed.

Studies on the Construction of Mutant Diversity Pool (MDP) lines, and their Genomic Characterization in Soybean

  • Dong-Gun Kim;Sang Hoon Kim;Chang-Hyu Bae;Soon-Jae Kwon
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2021년도 춘계학술대회
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    • pp.9-9
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    • 2021
  • Mutation breeding is useful for improving agronomic characteristics of various crops. In this study, we constructed soybean Mutant Diversity Pool (MDP) from 1,695 gamma-irradiated mutants through two selection phases over M1 to M12 generations; we selected 523 mutant lines exhibiting at least 30% superior agricultural characteristics, and, second, we eliminated redundant morphological phenotypes in the M12 generation. Finally, we constructed 208 MDP lines and investigated 11 agronomic traits. We then assessed the genetic diversity and inter-relationships of these MDP lines using target region amplification polymorphism (TRAP) markers. Among the different TRAP primer combinations, polymorphism levels and PIC values averaged 59.71% and 0.15, respectively. Dendrogram and population structure analyses divided the MDP lines into four major groups. According to an analysis of AMOVA, the percentage of inter-population variation among mutants was 11.320 (20.6%), whereas mutant inter-population variation ranged from 0.231 (0.4%) to 14.324 (26.1%). Overall, the genetic similarity of each cultivar and its mutants were higher than within other mutant populations. In an analysis of the genome-wide association study (GWAS) using based on the genotyping-by-sequencing (GBS), we detected 66 SNPs located on 13 different chromosomes were found to be highly associated with four agronomic traits: days of flowering (33 SNPs), flower color (16 SNPs), node number (6 SNPs), and seed coat color (11 SNPs). These results are consistent with those previously reported for other genetic resource populations, including natural accessions and recombinant inbred line. Our observations suggest that genomic changes in mutant individuals induced by gamma rays occurred at the same loci as those of natural soybean population. This study has demonstrated that the integration of GBS and GWAS can serve as a powerful complementary approach to gamma-ray mutation for the dissection of complex traits in soybean.

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당근 종모 형질 관련 cDNA Library 작성 및 EST 분석 (Construction of cDNA Library and EST Analysis Related to Seed-hair Characteristics in Carrot)

  • 오규동;심은조;전상진;박영두
    • 원예과학기술지
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    • 제31권6호
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    • pp.782-789
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    • 2013
  • 당근(Daucus carota L. var. sativa)은 세계적으로 널리 이용되는 작물 중 이며, 영양학적으로도 중요한 작물이다. 하지만, 종자 표피세포에서 생성되는 종자모는 발아를 억제하고 흡수를 저해하여 육묘에 어려움을 야기한다. 이러한 어려움을 타파하기 위해 당근 종자는 기계적인 제모작업을 거쳐 상품화 되고 있다. 이 과정에서 생산상의 여러 가지 단점들이 발생하며, 이를 보완하기 위해 단모종자 당근 품종의 육종이 필요하다. 따라서 본 연구는 단모종자 표현형 CT-ATR 615 OP 666-13개체와 장모종자 표현형 CT-ATR 615 OP 671-9개체 및 단모종자 표현형 CT-SMR 616 OP 659-1 개체와 장모종자 표현형 CT-SMR 616 OP 677-14개체 등 두 조합의 종자 cDNA library를 작성 후 EST 염기서열의 비교분석을 통해 당근 종자모 형질관련 연구에 이용하고자 하였다. 첫째로 EST 염기서열의 BlastX 결과를 바탕으로 각각의 EST를 FunCat 기능별 category로 분류하였다. 그 결과 Metabolism category와 protein folding 및 stabilization, protein binding, C-compound binding category에서 2조합 모두 동일한 유의적인 차이를 확인하였다. 두 번째로 EST 염기서열의 GO data를 바탕으로 seed trichome differentiation 및 cellulose biosynthetic process에 관련된 EST를 선발하였다. 이러한 FunCat category에서의 차이점과 GO data 분석을 통해 확인된 후보 EST 들이 당근 종자모 형성에 많은 영향을 미치는 것으로 생각된다. 마지막으로 분석된 개체 별 EST 염기서열을 바탕으로 33개의 SNP site, 741개의 SSR site를 확인하였다. 확인된 SNP 및 SSR site는 당근 종자모 형성에 관련된 분자마커 개발에 이용할 수 있음은 물론 당근의 여러 형질에 대한 연구에 활용 가능할 것으로 기대된다.