• Journal of Plant Biotechnology
• /
• 제43권2호
• /
• pp.157-163
• /
• 2016

본 연구는 국내 동, 서 및 남해안 바닷가 근처 사구지역에서 자생하는 번행초 55개체를 수집하여 AFLP 마커시스템을 적용하고 이들 유전자원들간의 유전다양성 차이를 알아보기 위하여 실시하였다. 우선 전체 게놈의 특이적 부위를 절단하기 위하여 제한효소로 EcoRI과 MseI 12개 조합을 활용하였고, 그 결과 총 1,279 절편을 확보할 수 있었다. 이 결과는 제한효소 조합당 평균 107개의 절편이 생산된 것으로 이 중 평균 62개(약 58%)가 유전자원간에 다형성을 나타냈다. 이와 같이 유전자원간에 다형성을 보인 게놈 절편을 대상으로 유전다양성을 분석한 결과 조사된 55개체 번행초 유전자원 집단은 29%의 유전적 차이를 보이는 것을 알 수 있었다. 또한 군집분석을 통해 유전적 차이를 보이는 그룹을 분류한 결과 국내 자생 번행초 유전자원은 총 7개의 집단으로 나누어짐을 알 수 있었다. 본 연구에서 국내외 최초의 번행초 유전 다양성 평가 정보는 향 후 품종 육성을 위한 교배친의 선발에 적용하여 다양한 유전적 차이를 보이는 분리집단을 확보하는 데 활용이 가능할 것으로 생각된다.

• Journal of Microbiology
• /
• 제40권4호
• /
• pp.254-259
• /
• 2002

The Phytophthora infestans requires two mating types for sexual reproduction. Amplified fragment length polymorphism (AFLP) was used to specifically detect different mating types of P. infestans. The AFLP primers E+AA (5'-GACTGCGTACCAATTCAA-3') and M+CAA (5'-GATGAGTCCTGAG-TAAC AA-3') detected a fragment that is specific in the A2 mating type of P. infestans. This fragment was cloned and sequenced. Based on the sequence data, PHYB-1 and PHYB-2 primer were designed to detect the A2 mating type of P. infestans. A single 347 bp segment was observed in the A2 mating type of P. infestans, but not in the A1 mating type of P. infestans or other Phytophthora spp. Identification of mating type was performed with phenotype (sexual reproduction) and genotype (CAPs marker) methods. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using mating type-specific primers, a unique band was obtained within annealing temperatures of 57$^{\circ}C$-62$^{\circ}C$ and DNA levels of 10pg-100 ng (data not shown).

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2001년도 추계 학술대회지
• /
• pp.146-147
• /
• 2001

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 1996년도 춘계 학술대회지
• /
• pp.28-29
• /
• 1996

• BMB Reports
• /
• 제36권5호
• /
• pp.427-432
• /
• 2003

Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

• 원예과학기술지
• /
• 제30권1호
• /
• pp.64-72
• /
• 2012

고추 역병은 그 동안 우리나라 고추 생산에 있어서 심각한 수량감소의 피해를 주어 왔으며, 2004년 처음으로 고추 역병 저항성 품종이 보급되기 시작하면서, 국내 건고추 육종에 있어서 역병 저항성의 도입은 필수불가결한 것으로 되었다. 그러므로 저항성 식물체 선발의 정확성 제고 및 육종의 효율 향상을 위하여 역병 저항성과 연관된 분자표지의 개발이 요구된다. 지금까지 고추 역병 저항성 주동 유전자에 연관된 분자표지가 일부 개발되어 있지만, 부분적으로 밖에 사용할 수 없다는 문제점이 있다. 따라서 본 연구에서는 역병 저항성 소재에 상관없이 저항성을 구별해 낼 수 있는 분자표지를 개발하고자 하였다. 이를 위해 '수비초' ${\times}$ 'CM334' 조합의 $F_2$ 분리집단($SF_2$)과 역병 저항성 시판 $F_1$품종('독야청청')을 자가수정한 $F_2$ 분리집단($DCF_2$)을 만들었다. BSA-AFLP 방법으로 총 1,024 프라이머 조합을 사용하여 역병 저항성과 연관된 세 개의 AFLP 분자표지(AFLP1, AFLP2 및 AFLP3)를 선발하였으며, 이를 CAPS 분자표지(M1-CAPS, M2-CAPS 및 M3-CAPS)로 전환하였다. 이 중 M3-CAPS 분자표지를 10개의 역병 저항성 계통, 14개의 이병성 계통, 'CM334'를 화분친으로 사용한 5개의 $F_1$ 조합, 그리고 53개의 시판 $F_1$ 품종에 적용해 본 결과, 역병 저항성 표현형과 M3-CAPS 분자표지의 마커형이 잘 일치하였고, P5-SNAP과 Phyto5.2-SCAR 분자표지보다 높은 실용성을 보이는 결과를 얻었다. 따라서, M3-CAPS 분자표지는 역병 저항성 고추 계통 육성에 많은 도움을 줄 수 있을 것으로 생각한다.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2001년도 춘계 학술대회지
• /
• pp.61-61
• /
• 2001

• 한국작물학회지
• /
• 제48권4호
• /
• pp.297-302
• /
• 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.

• 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.

• Mycobiology
• /
• 제32권3호
• /
• pp.123-127
• /
• 2004

Sweet persimmons have been increasingly cultivated in the southern part of Korea. However, anthracnose disease caused by Colletotrichum species is one of the major hindrances in cultivation and productions. In this study, we used polymerase chain reaction(PCR) to detect Colletotrichum species with the AFLP(amplified fragment length polymorphism) method. In AFLP, we used E3(5'-GACTGCGTACCAATTCTA-3') and M1(5'-GATGAGTCCTGAGTAACAG-3') primer combination and, as a result, 262 bp segment was observed in Colletotrichum species only. Specific PCR primers were designed from the sequence data and used to detect the presence of the fungus in genomic DNA isolated from symptomless sweet persimmon plants. Based on sequence data for specific segments, Co.B1(5'-GAGAGAGTAGAATTGCGCTG-3') and Co.B2(5'-CTACCATTCTTCTA GGTGGG-3') were designed to detect Colletotrichum species. The 220 bp segment was observed in Colletotrichum species only, but not in other fungal and bacterial isolates.