• 제목/요약/키워드: bulked segregant analysis

검색결과 25건 처리시간 0.022초

Molecular Marker Related to Fruitbody Color of Flammulina velutipes

  • Kong, Won-Sik;You, Chang-Hyun;Yoo, Young-Bok;Kim, Gyu-Hyun;Kim, Kwang-Ho
    • Mycobiology
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    • 제32권1호
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    • pp.6-10
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    • 2004
  • White and brown strains of Flammulina velutipes were inter-crossed. All $F_1$ showed light-brown fruitbody, suggesting that a gene for the brown fruitbody was incompletely dominant against the white one. And backcross experiment showed that more than two genes were involved in color determination. To isolate a molecular marker linked to fruitbody color, a set of primers was designed from a sequence of clones derived by a bulked segregant analysis. These markers showed a specific band which co-segregated with brown fruitbody forming strains.

Cleaved Amplified Polymorphic Sequence and Amplified Fragment Length Polymorphism Markers Linked to the Fertility Restorer Gene in Chili Pepper (Capsicum annuum L.)

  • Kim, Dong Sun;Kim, Dong Hwan;Yoo, Jae Hyoung;Kim, Byung-Dong
    • Molecules and Cells
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    • 제21권1호
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    • pp.135-140
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    • 2006
  • Cytoplasmic male sterility (CMS) in plants, which is due to failure to produce functional pollen, is a maternally inherited trait. Specific nuclear genes that suppress CMS, termed fertility restorer (Rf) genes, have been identified in several plants. In this study, Rfl-inked molecular markers in pepper (Capsicum annuum L.) were detected by bulked segregant analysis of eight amplified fragment length polymorphisms (AFLPs). Only AFRF8 was successfully converted to a cleaved amplified polymorphic sequence (CAPS) marker. This was named AFRF8CAPS and genotype determination using it agreed with that obtained with the original AFRF8. A linkage map with a total size of 54.1 cM was constructed with AFRF8CAPS and the seven AFLP markers using the Kosambi function. The AFRF8CAPS marker was shown to be closest to Rf with a genetic distance of 1.8 cM. These markers will be useful for fast and reliable detection of restorer lines during $F_1$ hybrid seed production and breeding programs in pepper.

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.

Resistance Potential of Bread Wheat Genotypes Against Yellow Rust Disease Under Egyptian Climate

  • Mahmoud, Amer F.;Hassan, Mohamed I.;Amein, Karam A.
    • The Plant Pathology Journal
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    • 제31권4호
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    • pp.402-413
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    • 2015
  • Yellow rust (stripe rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive foliar diseases of wheat in Egypt and worldwide. In order to identify wheat genotypes resistant to yellow rust and develop molecular markers associated with the resistance, fifty F8 recombinant inbred lines (RILs) derived from a cross between resistant and susceptible bread wheat landraces were obtained. Artificial infection of Puccinia striiformis was performed under greenhouse conditions during two growing seasons and relative resistance index (RRI) was calculated. Two Egyptian bread wheat cultivars i.e. Giza-168 (resistant) and Sakha-69 (susceptible) were also evaluated. RRI values of two-year trial showed that 10 RILs responded with RRI value >6 <9 with an average of 7.29, which exceeded the Egyptian bread wheat cultivar Giza-168 (5.58). Thirty three RILs were included among the acceptable range having RRI value >2 <6. However, only 7 RILs showed RRI value <2. Five RILs expressed hypersensitive type of resistance (R) against the pathogen and showed the lowest Average Coefficient of Infection (ACI). Bulked segregant analysis (BSA) with eight simple sequence repeat (SSR), eight sequence-related amplified polymorphism (SRAP) and sixteen random amplified polymorphic DNA (RAPD) markers revealed that three SSR, three SRAP and six RAPD markers were found to be associated with the resistance to yellow rust. However, further molecular analyses would be performed to confirm markers associated with the resistance and suitable for marker-assisted selection. Resistant RILs identified in the study could be efficiently used to improve the resistance to yellow rust in wheat.

Identification of Molecular Markers for Photoblastism in Weedy Rice

  • Lee, Hyun-Sook;Ahn, Sang-Nag;Sasaki, Kazuhiro;Chung, Nam-Jin;Choi, Kwan-Sam;Sato, Tadashi
    • 한국육종학회지
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    • 제42권2호
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    • pp.144-150
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    • 2010
  • The objective of this study was to map gene/QTL for photoblastism in a weedy rice (photoblastic rice: PBR) using DNA markers. Light-induced effect on germination of seeds was compared among three accessions (Oryza sativa L.), PBR, Milyang 23 and Ilpum. Results showed that PBR seeds started to show photoblastism during seed development, different from Ilpum and Milyang 23. Frequency distribution of germination in the F4 lines from crosses between Ilpum and PBR and, Milyang 23 and PBR revealed bimodal distributions suggesting that photoblastism was controlled by a few genes. Bulked segregant analysis using $F_4$ populations derived from the above two crosses was conducted to identify gene/QTL for photoblastism. Two QTL were identified on chromosomes 1 and 12 explaining 11.2 and 12.8% of the phenotypic variance, respectively. Two QTL were further mapped between two SSR markers, RM8260 and RM246 on chromosome 1, and between RM270 and 1103 on chromosome 12. It is noteworthy that two QTL for photoblastism were colocalized with the QTL for seed dormancy reported in the previous QTL studies. The clustering of two genes for photoblastism and dormancy possibly indicates that these regions constitute rice phytochrome gene clusters related to germination. Because PBR has a low degree of dormancy, a pleiotropic effect of a single gene controlling dormancy and photoblastism can be ruled out. The linked markers will provide the foundation for positional cloning of the gene.

베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발 (Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan)

  • 김수철;김혜수;조용운;류재산;조수정
    • 한국버섯학회지
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    • 제13권1호
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    • pp.79-83
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    • 2015
  • 본 연구는 큰느타리버섯의 베타글루칸 고함유 형질에 관련된 SCAR marker를 개발하기 위해 수행되었다. operon사의 OPA(20개), OPB(20개), OPL(20개), OPP(20개), OPR(20개), OPS(20개) 등 총 120개 primer를 random primer(10 mer)로 사용하여 대립 계통 9종과 베타글루칸 고함유 계통 9 종을 대상으로 RAPD를 이용한 bulked segregant analysis를 실시하여 OP-R03 primer로부터 대립 계통에는 나타나지 않고 베타글루칸 고함유 계통에만 나타나는 특이적인 RAPD 밴드를 얻었다. OP-R03 primer를 이용한 RAPD 결과, 약 91 bp 부근에서 베타글루칸 고함유 계통에 특이적인 DNA 밴드가 관찰되었으며 이 DNA 밴드의 염기서열 말단을 근거로 SCAR 마커로 사용할 specific primer인 OP-R03-1-F와 OP-R03-1-R를 디자인하였다. SCAR 마커 OP-R03-1-F/-1-R primer를 이용하여 PCR을 수행한 결과에서도 91 bp 부근에서 대립 계통과 구별되는 DNA 밴드가 베타글루칸 고함유 계통에서 확인되었으며 random primer인 OP-R03 primer를 이용하여 PCR을 수행했을 때보다 재현성이 높고 진한 DNA 밴드임을 확인할 수 있었다.

큰느타리버섯의 고온적응성 형질에 관련된 SCAR Marker 개발 (Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains adaptable to high-temperature)

  • 김수철;김혜수;박소연;류재산;조수정
    • 한국버섯학회지
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    • 제12권3호
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    • pp.226-231
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    • 2014
  • 본 연구는 큰느타리버섯의 고온적응성 형질에 관련된 SCAR marker를 개발하기 위해 수행되었다. operon 사의 OPA(20개), OPB(20개), OPL(20개), OPP(20개), OPR(20개), OPS(20개) 등 총 120개 primer를 random primer(10 mer)로 사용하여 대립 계통 7종과 고온성 계통 7 종을 대상으로 RAPD를 이용한 bulked segregant analysis를 실시하여 OP-A06 primer로부터 대립 계통에는 나타나지 않고 고온성 계통에만 나타나는 특이적인 RAPD 밴드를 얻었다. OP-A06 primer를 이용한 RAPD 결과, 약 385 bp 부근에서 고온성 계통에 특이적인 DNA 밴드가 관찰되었으며 이 DNA 밴드의 염기서열 말단을 근거로 SCAR 마커로 사용할 specific primer인 OP-A06-1-F와 OP-A06-1-R를 디자인하였다. SCAR 마커 OP-A06-1-F/-1-R primer를 이용하여 PCR을 수행한 결과에서도 385 bp 부근에서 대립 계통과 구별되는 DNA 밴드가 고온성 계통에서 확인되었으며 random primer인 OP-A06 primer를 이용하여 PCR을 수행했을 때보다 재현성이 높고 진한 DNA 밴드임을 확인할 수 있었다.

Detection of Individuals Restoring Fertility by DNA Fragment Converted into STS (sequence tagged site) in Red pepper

  • Lee, Jang-Soo;Lee, Keun-Hyang;Kim, Yeo-Song;Kang, Kwon-Kyoo;Nou, Ill-Sup;Hirata, Yutaka
    • Plant Resources
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    • 제7권2호
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    • pp.136-140
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    • 2004
  • Co-segregation of male fertility with DNA markers selected by RAPD analysis as being potentially linked to the restorer gene (Rf) for Cytoplasmic male sterility (CMS) was analyzed using segregating F2 population. One RAPD marker directly linked to the Rf locus was identified. Amplification of OPT-02/570 using the STS primers generated a monomorphic band of each fertile plants randomly selected F2 progenies. From these results, this specific marker would be strongly linked to be restoring gene. The use of STS marker is effective in overcoming the reliability of the RAPD phenotype and improving their utility for MAS, co-dominant STS markers are especially very useful.

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Construction of a Genetic Linkage Map of Shiitake Mushroom Lentinula Edodes Strain L-54

  • Hoi-Shan, Kwan;Hai-Lou, Xu
    • BMB Reports
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    • 제35권5호
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    • pp.465-471
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    • 2002
  • From fruiting bodies of L. edodes strain L-54, single-spore isolates (SSIs) were collected. Two parental types of L-54 were regenerated via monokaryotization. By means of random-amplified polymorphic DNA (RAPD), DNA samples from L-54, its two parental types, and 32 SSIs were amplified with arbitrary primers. Dedikaryotization was demonstrated, and 91 RAPD-based molecular markers were generated. RAPD markers that were segregated at a 1:1 ratio were used to construct a linkage map of L. edodes. This RAPD-linkage map greatly enhanced the mapping of other inheritable and stable markers [such as those that are linked to a phenotype (the mating type), a known gene (priA) and a sequenced DNA fragment (MAT)] with the aid of mating tests, bulked-segregant analysis, and PCR-single-strand conformational polymorphism. These markers comprised a genetic map of L. edodes with 14 linkage groups and a total length of 622.4 cM.