• Molecules and Cells
• /
• 제21권1호
• /
• pp.135-140
• /
• 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.

• 한국작물학회지
• /
• 제49권5호
• /
• pp.429-433
• /
• 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.

• The Plant Pathology Journal
• /
• 제31권4호
• /
• pp.402-413
• /
• 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.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2017년도 9th Asian Crop Science Association conference
• /
• pp.102-102
• /
• 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.

• Molecules and Cells
• /
• 제26권6호
• /
• pp.548-553
• /
• 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.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
• /
• pp.50-50
• /
• 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.

• 한국버섯학회지
• /
• 제13권1호
• /
• pp.79-83
• /
• 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.

• 한국버섯학회지
• /
• 제12권3호
• /
• pp.226-231
• /
• 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.

• BMB Reports
• /
• 제39권1호
• /
• pp.37-45
• /
• 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.

• 생명과학회지
• /
• 제21권12호
• /
• pp.1659-1665
• /
• 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.