• 제목/요약/키워드: bulked segregant analysis

검색결과 25건 처리시간 0.034초

배추무사마귀병 저항성 유전자와 연관된 DNA 마커개발 (Development of DNA markers linked to resistant gene to Psmodiophora brassicae Woronin in Chinese cabbage)

  • 한영한;우종규;박철호
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
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    • pp.50-50
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    • 2002
  • 배추무사마귀병 저항성 유전 양식을 증명하기 위해서 CR계 F1에서 유래된 F2 세대를 포장시험과 유묘 검정을 실시하였다. F$_2$ 세대의 7 집단은 단인자우성으로 3:1의 분리비를 보였고, 5 집단은 중복 유전자가 관여하는 9:7의 유전 분리비를 보였다. 배추무사마귀병 저항성 유전자와 연관된 DNA 마커를 개발하기 위하여 CR-Saerona F$_2$ 집단을 배추무사마귀병 발병포장에서 재배하여 저항성 평가를 하였다. 220개의 임의의 프라이머를 이용하여 BSA-RAPD (Bulked segregant analysis-Randomly amplified polymorphic DNA)를 수행하였지만 CR-Saerona F2 집단에서 배추무사마귀병 저항성 유전자와 꼭 들어맞는 DNA 마커는 발견되지 않았다. 300개의 임의의 프라이머를 이용하여 CR-Saerona에서 유래된 F$_2$ 세대를 QTL 분석하였다. 저항성 정도는 발병지수에 따라 조사되었고 QTL 분석을 위해 one-way ANOVA 테스트를 하였다. 통계분석 결과 두 프라이머(K16-1, L2-2)가 저항성과의 상관관계를 보여 주었으나 유의성은 인정되지 않았다.

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Identification of Sex-specific Expression Markers in the Giant Tiger Shrimp (Penaeus monodon)

  • Khamnamtong, Bavornlak;Thumrungtanakit, Supaporn;Klinbunga, Sirawut;Aoki, Takashi;Hirono, Ikuo;Menasveta, Piamsak
    • BMB Reports
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    • 제39권1호
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    • pp.37-45
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    • 2006
  • Bulked segregant analysis (BSA) and AFLP were used for isolation of genomic sex determination markers in Penaeus monodon. A total of 256 primer combinations were tested against 6-10 bulked genomic DNA of P. monodon. Five and one candidate female- and male-specific AFLP fragments were identified. Female-specific fragments were cloned and further characterized. SCAR markers derived from FE10M9520, FE10M10725.1, FE10M10725.2 and FE14M16340 provided the positive amplification product in both male and female P. monodon. Further analysis of these markers using SSCP and genome walk analysis indicated that they were not sex-linked. In addition, sex-specific (or differential) expression markers in ovaries and testes of P. monodon were analyzed by RAP-PCR (150 primer combinations). Twenty-one and fourteen RAP-PCR fragments specifically/differentially expressed in ovaries and testes of P. monodon were successfully cloned and sequenced. Expression patterns of 25 transcripts were tested against the first stranded cDNA of ovaries and testes of 3-month-old and broodstock-sized P. monodon (N = 5 and N = 7 - 10 for females and N = 4 and N = 5 - 7 for males, respectively). Five (FI-4, FI-44, FIII-4, FIII-39 and FIII-58) and two (M457-A01 and MII-51) derived RAP-PCR markers revealed female- and male-specific expression patterns in P. monodon. Surprisingly, MII-5 originally found in testes showed a higher expression level in ovaries than did testes of juvenile shrimps but a temporal female-specific pattern in P. monodon adults.

'진귤' (Citrus sunki) 의 웅성가임 연관 SCAR 마커 개발 (Development of a SCAR Marker Linked to Male Fertility Traits in 'Jinkyool' (Citrus sunki))

  • 채치원;;윤수현;박재호;이동훈
    • 생명과학회지
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    • 제21권12호
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    • pp.1659-1665
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    • 2011
  • 감귤류 중 수술이 퇴화되어 웅성불임형질을 나타내는 '청견' 품종에 정상적인 수술의 형태를 가진 웅성가임인 '진귤' 품종을 교배하여 150개체의 $F_1$ 집단을 구축하여 수술이 퇴화되는 개체와 정상인 개체를 분리하였다. 분리된 F1 개체들을 사용하여 SRAP 기법과 집단 분리 분석법(BSA)을 조합하여 웅성 가임 연관 마커 개발에 활용하였다. $F_1$ 집단 내 150개체 중 66개체가 퇴화 수술을 갖고 있으며 웅성 가임성과 웅성 불임성의 분리비는 1:1이며 $x^2$ 값은 2.16(p=0.05)이었다. 197개의 SRAP 프라이머 조합들 중 웅성가임 특이밴드를 형성하는 3개의 SRAP 프라이머 조합(F4/R27, F39/R60, 및 F15/R37)을 선발하였으며, 이 중 F39/R60 프라이머에 특이적으로 증폭하는 DNA단편의 염기서열을 기본으로 하여 새롭게 작성한 양방향 프라이머 조합 중 웅성 가임 계통에서만 약 1.4 Kb의 특이밴드를 증폭하는 프라이머 조합, pMS 33U/pMS 1462L를 선발하여 SCAR 마커를 개발 하였다. 이러한 결과는 개발된 SCAR 마커로 무핵성 계통들의 육종 선발에 효율성을 높일 수 있을 것으로 기대된다.

딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발 (Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria×ananassa Duchesne))

  • 제희정;안재욱;윤혜숙;김민근;류재산;홍광표;이상대;박영훈
    • 원예과학기술지
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    • 제33권5호
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    • pp.722-729
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    • 2015
  • 딸기 흰가루병은 Podosphaera aphanis에 의해 발병되며 수확기에 가장 큰 피해를 주는 병으로 현재 유황, 농약으로 주로 방제 되고 있는 실정이다. 본 연구에서는 딸기 흰가루병 저항성 품종 육성을 위한 흰가루병 저항성 특이마커 개발로 내병성 육종효율을 높이고자 하였다. 흰가루병 저항성 계통 선발을 위한 분자마커를 개발하기 위해 아키히메${\times}$설향 집단을 대상으로 자가수분을 통해 후대 양성 후 병저항성을 검정하였다. 마커분석은 RAPD primer 200 세트 중 OPE10 331bp에서부터 흰가루병 저항성 특이 마커 선발하였다. 흰가루병 저항성 특이밴드만 선발하기 위하여 클로닝 후 유전자정보 분석하여 SP1F/R의 Primer를 제작하였다. 그러나 SP1F/R을 이용하여 PCR한 결과 저항성, 감수성간에 다형성이 확인되지 않아 염기서열을 정렬한 후 SNP, In/del의 다형성 유무를 확인한 결과 6개의 SNP를 확인하였다. 이들 PCR 산물을 해당 사이트와 연관된 제한효소로 절단한 결과 그 중 Eae I(Y/GGCCR)의 절단으로 231bp 위치에서 저항성과 감수성간의 다형성을 확인함으로써 흰가루병 저항성 계통선발을 위한 분자마커를 선발하였다. 이러한 과정을 통해 딸기 흰가루병 저항성 품종 육성을 위한 MAS(marker assisted selection) 체계 확립으로 내병성 육종효율 증진에 기여를 할 수 있을 것으로 기대된다.

Analysis of 'QTL-seq' associated with allelopathic potential in rice

  • Cho, Gi-Won;Choi, Ji-Su;Oh, Young-Taek;Lee, Kyoung-Jin;Chung, Ill-Min
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.102-102
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    • 2017
  • In this study, QTL analysis of allelopathy was conducted. A total of 171 of F8 RILs developed from the cross between Nongan(low allelopathic cultivar) and Sathi(high allelopathic cultivar) were used . the performance of allelopathy were evaluated using 'ECAM(Equal Compartment Agar Method)', where the root length of lettuce cultivated with the RILs were measured. The distribution of the performance was followed as normal distribution. In order to identify the location of QTLs related to allelopathy, QTL-seq with BSA(Bulked-segregant analysis) was performed with 20 highest and 10 lowest RILs. As a result, Two Sliding window coordinate region of candidate QTLs were detected on Chr4 (5,050,001 - 14,800,000, 18,650,001 - 22,500,000), Chr8 (2,550,001 - 8,250,000, 21,150,001 - 26,800,000) and One region on Chr7 (1 - 3,300,000), Chr9 (1 - 13,300,000) respectively.

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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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    • 제26권6호
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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.

Genetic analysis of clubroot resistance in Chinese cabbage using single spore isolate of Plasmodiophora brassicae and development of RAPD marker linked to its resistance gene

  • Cho, Kwang-Soo;Hong, Su-Young;Han, Young-Han;Yoon, Bong-Kyeong;Ryu, Seoung-Ryeol;Woo, Jong-Gyu
    • Journal of Crop Science and Biotechnology
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    • 제11권2호
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    • pp.101-106
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    • 2008
  • To identify inheritance of clubroot disease resistance genes in Chinese cabbage, seedling tests of $BC_1P_1,\;BC_1P_2$, and $F_2$ populations derived from $F_1$ hybrid(var. CR Saerona) using single spore isolate(race 4 identified with William's differential host) from Plasmodiophora brassciae were conducted. Resistance(R) and susceptible(S) plants segregated to 1:0 in backcross to the resistant parent. The $F_2$ population segregated in a 3(R):1(S) ratio. This result implied that the resistance of clubroot disease is controlled by a single dominant gene to the race 4 of P. brassicae in CR Saerona. To develop DNA markers linked to clubroot resistance genes, 185 plants of CR Saerona among $F_2$ populations were used. A total of 300 arbitrary decamer was applied to $F_2$ population using BSARAPD(Bulked segregant analysis-Randomly amplified polymorphic DNA). One RAPD marker linked to clubroot resistance gene in CR Saerona($OPJ_{1100}$) was identified. This marker was 3.1 cM in distance from resistance gene in $F_2$ population. This marker may be useful for a marker-assisted selection(MAS) and gene pyramiding of the clubroot disease resistant gene in Chinese cabbage breeding programs.

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Development of a Sequence Characteristic Amplified Region Marker linked to the L4 Locus Conferring Broad Spectrum Resistance to Tobamoviruses in Pepper Plants

  • Kim, Hyun Jung;Han, Jung-Heon;Yoo, Jae Hyoung;Cho, Hwa Jin;Kim, Byung-Dong
    • Molecules and Cells
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    • 제25권2호
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    • pp.205-210
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    • 2008
  • To develop molecular markers linked to the $L^4$ locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic $BC_4F_2$ generation for the $L^4$ locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a $BC_{10}F_2$ derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 $F_2$ T102 individuals showed that they were each within 2.5 cM of the $L^4$ locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the $L^4$ locus in T102 and 0.9 cM in another $BC_{10}F_2$ population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time.

Genetic analysis of photoblastism and mesocotyl elongation

  • Lee, Hyun-Sook;Kang, Ju-Won;Jeon, Yun-A;Ahn, Sang-Nag
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.26-26
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    • 2017
  • Seed germination stimulated by light is said to be photoblastism. Photoblastism has not been reported in cereal crops, especially in the rice, but Korean weedy rice was reported to have photoblastism and longer mesocotyl than cultivar. Photoblastic weedy rice (PBR) was used to identify QTLs for photoblastism and mesocotyl length. In previous works, QTLs for photoblastism, pbr1 and pbr12 were identified on chromosomes 1 and 12 using 124 F4 lines from a cross between Ilpum and PBR using bulked segregant analysis. Two QTLs for mesocotyl elongation, qMel-1 and qMel-3 were mapped on chromosomes 1 and 3 120 F8 lines from the same cross. Of interest, the RM8260-RM246 region of pbr1 overlapped with a region of qMel-1. To know whether these two QTLs are functionally related, 110 F3 lines were developed from a cross between Ilpum and CR7124. CR7124 having photoblastism and long mesocotyl was selected from 120 F8 lines. 95 F3 lines were measured for germination rate in a light and dark condition and mesocotyl length. Mesocotyl length and germination rate in the dark condition in F3 lines showed significant correlation (r = 0.7, P < 0.0001). 95 $F_3$ lines were genotyped with RM7419 on chromosome 1. ANOVA showed that RM7419 was tightly linked to QTLs for photoblastism as well as mesocotyl length on chromosome 1 (P < 0.0001) indicating the tight linkage of two QTLs. Fine mapping of the two QTL is underway to analyze their functional relationship.

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Identification of Coupling and Repulsion Phase DNA Marker Associated With an Allele of a Gene Conferring Host Plant Resistance to Pigeonpea sterility mosaic virus (PPSMV) in Pigeonpea (Cajanus cajan L. Millsp.)

  • Daspute, Abhijit;Fakrudin, B.
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.33-40
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    • 2015
  • Pigeonpea Sterility Mosaic Disease (PSMD) is an important foliar disease caused by Pigeonpea sterility mosaic virus (PPSMV) which is transmitted by eriophyid mites (Aceria cajani Channabasavanna). In present study, a F2 mapping population comprising 325 individuals was developed by crossing PSMD susceptible genotype (Gullyal white) and PSMD resistant genotype (BSMR 736). We identified a set of 32 out of 300 short decamer random DNA markers that showed polymorphism between Gullyal white and BSMR 736 parents. Among them, eleven DNA markers showed polymorphism including coupling and repulsion phase type of polymorphism across the parents. Bulked Segregant Analysis (BSA), revealed that the DNA marker, IABTPPN7, produced a single coupling phase marker (IABTPPN $7_{414}$) and a repulsion phase marker (IABTPPN $7_{983}$) co-segregating with PSMD reaction. Screening of 325 F2 population using IABTPPN7 revealed that the repulsion phase marker, IABTPPN $7_{983}$, was co-segregating with the PSMD responsive SV1 at a distance of 23.9 cM for Bidar PPSMV isolate. On the other hand, the coupling phase marker IABTPPN $7_{414}$ did not show any linkage with PSMD resistance. Additionally, single marker analysis both IABTPPN $7_{983}$ (P<0.0001) and IABTPPN $7_{414}$ (P<0.0001) recorded a significant association with the PSMD resistance and explained a phenotypic variance of 31 and 36% respectively in $F_2$ population. The repulsion phase marker, IABTPPN7983, could be of use in Marker-Assisted Selection (MAS) in the PPSMV resistance breeding programmes of pigeonpea.