• Title/Summary/Keyword: AFLP marker

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Development of a SCAR Marker Linked to Ph-3 in Solanum ssp.

  • Park, Pue Hee;Chae, Young;Kim, Hyun-Ran;Chung, Kyeong-Ho;Oh, Dae-Geun;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.139-143
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    • 2010
  • Late blight caused by Phytophthora infestans is historically a serious epidemic disease in potato and tomato cultivations. Accession L3708 (Solanum pimpinellifolium), a new source for late blight resistance was identified in AVRDC, and carries the resistance gene, Ph-3, incompatible to P. infestans race 3. The AFLP markers linked to Ph-3 were previously developed from the L3708 accession (Chunwongse et al. 2002). To facilitate tomato breeding with the Ph-3 gene, an attempt was made to convert AFLP markers to sequence-characterized amplified region (SCAR) markers. Among 6 AFLP markers, only one AFLP marker, L87, was successfully converted to SCAR marker. The resistance-specific 230 bp AFLP fragment was cloned and sequenced, and the PCR primer amplifying a 123 bp fragment was designed. This SCAR marker could discriminate resistant and susceptible individuals with high stringency. The developed SCAR marker could be used for the marker assisted-selection in tomato breeding programs.

Identification of Korean Native Goat Meat using Amplified Fragment Length Polymorphism (AFLP) DNA Markers (Amplified Fragment Length Polymorphism (AFLP) DNA Marker를 이용한 한국 재래흑염소육 감별)

  • 정의룡
    • Food Science of Animal Resources
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    • v.22 no.4
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    • pp.301-309
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    • 2002
  • This study was carried out to develop the breed-specific DNA markers for breed identification of Korean native goat meat using amplified fragment length polymorphism (AFLP)-PCR techniques. The genomic DNAs of Korean native goat, imported black goat and four dairy goat breeds(Saanen, Alpine, Nubian and Toggenburg) were extracted from muscle tissues or blood. Genomic DNA was digested with a particular combination of two restriction enzymes with 4 base(Mse I and Taq I) and 6 base(EcoR I and Hind III) recognition sites, ligated to restriction specific adapters and amplified using the selective primer combinations. In AFLP profiles of polyacrylamide gels, the number of scorable bands produced per primer combination varied from 36 to 74, with an average of 55.5. A total of 555 bands were produced, 149(26.8%) bands of which were polymorphic. Among the ten primer combinations, two bands with 2.01 and 1.26 kb in M13/H13 primer and one band with 1.65 kb in E35/H14 primer were found to be breed-specific AFLP markers in Korean native goat when DNA bands were compared among the goat breeds. In the E35/H14 primer combination, 2.19, 2.03, 0.96 and 0.87 kb bands detected in imported black goat, 2.13 kb band in Saanen breed and 2.08 kb band in Nubian breed were observed as breed-specific bands showing differences between goat breeds, respectively. The E35/H14 primer combination produced four DNA bands distinguished between Korean native goat and Saanen breed. The is study suggested that the breed specific AFLP bands could be used as DNA markers for the identification of Korean native goat meat from imported black goat and dairy goat meats.

Diversity and Inheritance of AFLP Markers in Wild and Cultivated Soybeans (AFLP marker를 이용한 콩의 유전적 다양성과 유전분리 분석)

  • 김용호;윤홍태
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.265-271
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    • 2004
  • Genetic variation is the basis of crop improvement. Limited genetic diversity in a crop species may restrict the amount of genetic improvement that can be achieved through plant breeding. Soybean is one of the world's most important crops. A potential source of genetic variability for the cultivated soybean is the wild species G. soja Sieb. & Zucc. Amplified fragment length polymorphism (AFLP) analysis is a PCR-based technique, which can detect a 10-fold greater nubmer of loci than other DNA marker analysis. Twenty cultivated soybeans and two-hundred wild soybeans were used to determine genetic vatiations by AFLPs and evaluate the usefulness of AFLPs as DNA markers. Six-hundred and ten fragments were detected with an average of 56 AFLP fragments produced per primer in a total of 11 AFLP primer pairs. The number of polymorphic loci detected per primer ranged from 7 to 20 and the polymorphism was greater in wild than in cultivated soybean. F$_2$ segregation analysis of four AFLP fragments in combination of Hwaeomputkong ${\times}$ PI 417479 indicated that they segregate as stable Mendelian loci with 3 : 1. This results strongly suggest that the AFLP analysis is a good technique for the detection of genetic polymorphism in a wide plant species.

Identification of AFLP Marker Linked to a SCN Resistant Gene in Soybean

  • Ko, Mi-Suk;Kim, Myung-Sik;Han, Soung-Jin;Chung, Jong-Il;Kang, Jin-Ho
    • Plant Resources
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    • v.5 no.3
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    • pp.169-175
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    • 2002
  • The soybean cyst nematode (Heterodera glycines Inchinoe; SCN) is a devastating pest of soybean and is responsible for significant losses in yield. The use of resistant cultivars is the effective method to reduce or eliminate SCN damage. The objective of this research is to identify AFLP markers linked to the SCN resistant genes. Bulked genomic DNA was made from resistant and susceptible genotypes to SCN and a total of 19 primer combinations were used. About 31 fragments were detected per primer combination. The banding patterns were readily distinguished in resistant and susceptible bulked genotypes. Polymorphic fragments were detected between resistant and susceptible bulked genotypes in the primer combination of CGT/GGC, CAG/GTG and CTC/GAG. In primer combinations of CGT/GGC and CAG/GTG, bulked resistant genotype produced a polymorphic bands. However, in primer of CTC/GAG, bulked susceptible genotype produced a polymorphic fragments. Three AFLP markers identified as a polymorphic fragments between bulked genomic DNA were mapped in 85 F2 population. Among them, only two markers, CGT/GGC and CTC/GAG, was linked and was mapped. Broad application of AFLP marker would be possible for improving resistant cultivars to SCN.

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Discrepancies between Mitochondrial DNA and AFLP Genetic Variation among Lineages of Sea Slaters Ligia in the East Asian Region

  • Kang, Seunghyun;Jung, Jongwoo
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.4
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    • pp.347-353
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    • 2020
  • Although sea slaters Ligia have a significant role in rocky shore habitats, their taxonomic entities have not been clearly understood. In this study, we investigated whether genetic variation inferred from a nuclear genetic marker, namely amplified fragment length polymorphism (AFLP), would conform to that of a mitochondrial DNA marker. Using both the mitochondrial DNA marker and the AFLP marker amplified by the six selective primer sets, we analyzed 95 Ligia individuals from eight locations from East Asia. The direct sequencing of mitochondrial 16S rRNA gene revealed three distinct genetic lineages, with 9.8-11.7 Kimura 2-parameter genetic distance. However, the results of AFLP genotyping analysis with 691 loci did not support those of mitochondrial DNA, and revealed an unexpectedly high proportion of shared polymorphisms among lineages. The inconsistency between the two different genetic markers may be explained by difference in DNA evolutionary history, for example inheritance patterns, effective population size, and mutation rate. The other factor is a possible genomic island of speciation, in that most of the genomic parts are shared among lineages, and only a few genomic regions have diverged.

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
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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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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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.

An AFLP-based Linkage Map of Japanese Red Pine (Pinus densiflora) Using Haploid DNA Samples of Megagametophytes from a Single Maternal Tree

  • Kim, Yong-Yul;Choi, Hyung-Soon;Kang, Bum-Yong
    • Molecules and Cells
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    • v.20 no.2
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    • pp.201-209
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    • 2005
  • We have constructed an AFLP-based linkage map of Japanese red pine (Pinus densiflora Siebold et Zucc.) using haploid DNA samples of 96 megagametophytes from a single maternal tree, selection clone Kyungbuk 4. Twenty-eight primer pairs generated a total of 5,780 AFLP fragments. Five hundreds and thirteen fragments were verified as genetic markers with two alleles by their Mendelian segregation. At the linkage criteria LOD 4.0 and maximum recombination fraction 0.25(${\theta}$), a total of 152 markers constituted 25 framework maps for 19 major linkage groups. The maps spanned a total length of 2,341 cM with an average framework marker spacing of 18.4 cM. The estimated genome size was 2,662 cM. With an assumption of equal marker density, 82.2% of the estimated genome would be within 10 cM of one of the 230 linked markers, and 68.1% would be within 10 cM of one of the 152 framework markers. We evaluated map completeness in terms of LOD value, marker density, genome length, and map coverage. The resulting map will provide crucial information for future genomic studies of the Japanese red pine, in particular for QTL mapping of economically important breeding target traits.