• The Plant Pathology Journal
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• 제22권1호
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

• Fisheries and Aquatic Sciences
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• 제6권1호
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• pp.13-19
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• 2003

Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

• 한국작물학회지
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• 제46권4호
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• pp.341-343
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• 2001

Molecular markers have become fundamental tools for crop genome study. The objective of this study was to construct a genetic linkage map for cowpea with PCR-based molecular markers. Five hundred and twenty random RAPD primers were screened for parental polymorphism. Ninety RAPD markers from sixty primers was segregated in 75 F2 mapping population derived from the cross of local cultivars GSC01 and GSC02. 70 RAPD markers were found to be genetically linked and formed 11 linkage groups. Linkage map spanned 474.1 cM across all 11 linkage groups. There are six linkage groups of 40 cM or more, and five smaller linkage groups range from 4.9 to 24.8 cM. The average linkage distance between pairs of markers among all linkage groups was 6.87 cM. The number of markers per linkage group ranged from 2 to 32. The longest group 1 spans 190.6 cM, while the length of shortest group 11 is 4.9 cM. This map is further needed to be saturated with the various markers such as RFLP, AFLP, SSR and more various populations and primers. In addition, morphological markers and biochemical markers should be united to construct a comprehensive linkage map.

• Animal cells and systems
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• 제5권4호
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• pp.333-339
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• 2001

Common carp (Cyprinus carpio) and its aquaculture breed Israeli carp samples were obtained from two separate aquaculture facilities under the similar raising conditions during two years in the Kunsan National University, Korea. Genomic DNA was isolated from the common carp and Israeli carp for identification of genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA using arbitrary primers. The arbitrary primer No.21 (ACTTCGCCAC) yielded the highest number of fragments with the average of 15.0 among the primers used in Israeli carp. A tota1 of 294 polymorphic products in common carp and 336 in Israeli carp were observed by random primers. The average number of polymorphic products generated by random RAPD primer No. 2 (GTAGAC-CCGT) showed 8.0 in Israeli carp. On average, each random RAPD primer produced 5.4 amplified polymorphic products in common carp and 6.2 in Israeli carp. An average genetic similarity (BS value) was 0.44$\pm$0.05 within the common carp and 0.32$\pm$0.04 within the Israeli carp. The degree of similarity frequency (BS) between two carps was 0.67 as generated by the primer No. 19 (GACGGATCAG). The average level of bandsharing was 0.57$\pm$0.03 between the two carps. Accordingly, the two carp populations were genetically a little distant. The electrophoretic analysis of PCR-RAPD products showed middle levels of variation between the two carp populations. This result implies that the genetic diversity among intra-population may be higher when compared with that between the two carps. The RAPD polymorphism generated by these random primers might be used as a genetic marker for populations or lines identification in important aquacultural carp.

• Plant Biotechnology Reports
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• 제4권1호
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• pp.1-7
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• 2010

Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

• 아시안잔디학회지
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• 제23권2호
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• pp.307-316
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• 2009

크리핑벤트그래스는 골프코스의 퍼팅그린에서 주로 사용하고 있는 한지형잔디 초종이다. 크리핑벤트그래스 품종들은 형태적인 특성이나 유전적인 다양성이 협소하여 품종간 구분이 매우 어렵다. 따라서 이들 품종간 유전적인 다양성이나 골프코스 그린의 인터씨딩율에 대한 조사를 위해 SCAR marker를 개발하고자 본 연구를 수행하였다. penncross, penn A-4, crenshaw, L-93, CY-2, T-1 공시품종들에 대한 SCAR marker 개발을 위해 5개 RAPD primer에 대한 품종 간 구분이 가능한 특이적 band를 선발하였다. 선발한 특이적 band의 염기서열을 분석하여 품종별 SCAR primer를 제작하였다. 각 품종의 SCAR primer별로 특이적 band가 나타나는지에 대한 여부를 검정한 결과, penncross, penn A-4, crenshaw 품종의 SCAR primer는 6개 공시품종에서 동일한 크기의 band가 나타나 SCAR marker로써 활용이 어려웠다. 하지만 CY-2의 CY850F/R primer는 850bp, T-1의 T700F/R primer는 700bp, L-93의 L2900F/R는 2.9kb에서 특이적 band가 나타나 3개 품종별 SCAR marker로서 활용이 가능하였다. 따라서 본 연구에서 개발한 3개 품종별 SCAR marker를 활용하여 크리핑벤트그래스의 품종별 유전적 다양 및 골프코스 그린의 인터씨딩율 조사에 활용이 가능할 것으로 기대된다.

• 한국약용작물학회지
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• 제15권1호
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• pp.1-5
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• 2007

청도라지와 백도라지를 DNA 수준에서 구분하기 위하여, RAPD 분석을 바탕으로 SCAR primer (SPgR1, SPgR2)를 제작하여 이를 적용한 실험 결과는 다음과 같다. 총 24개의 임의 프라이머를 적용하여 6개의 청도라지와 백도라지 특이적인 프라이머를 선발하였고, 이들 프라이머를 이용하여 PCR을 수행한 결과 총 18개의 다형성을 나타내는 DNA fragment를 확보하였다. 확보된 DNA fragment 중 2개에 대한 염기서열 분석을 수행한 결과, 청도라지 특이적인 DNA 밴드는 887 bp의 염기서열로 구성되어 있었고 백도라지는 863 bp의 염기서열로 구성되어 있었다. 이들 염기서열을 바탕으로 두개의 SCAR primer를 제작하였고, 이중 백도라지 특이적인 SPgR2를 이용하여 PCR 한 결과, 청도라지에서는 약 500 bp 크기에서 두 개의 특이적인 밴드가 형성되었으며, 백도라지의 경우 한 개의 특이적인 밴드만 관찰되어 청도라지와 백도라지를 구분할 수 있었다. 결론적으로 본 실험을 통하여 개발된 SCAR 마커는 청도라지와 백도라지를 구분하기에 유용함을 확인하였다.

• 한국산림과학회지
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• 제87권3호
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• pp.422-428
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• 1998

구상나무 개체목으로부터 채취한 48개의 배유조직을 이용해서 PCR 방법에 의해 생성된 inter-simple sequence repeats(I-SSR) 표지자를 분석했다. 예비실험에서 6개의 배유조직을 이용해서 35개의 primer를 검색했으며, 그들 중에서 PCR 반응이 가장 잘되는 19개 primer를 선정해서 48개 배유조직을 이용한 본 실험에 사용했다. 카이자승 검정 결과, 19개 primer에 의해 증폭된 51개의 증폭산물이 5% 유의 수준에서 멘델의 분리비(1:1)에 따라 차대에 유전됨을 확인할 수 있었다. 멘델 유전자좌로 확인된 51개 표지자들의 게놈내 분포양상을 확인하기 위해서 연관분석을 수행한 결과, 51개 유전자좌들이 상호간에 서로 연관되어있지 않은 것으로 확인되어 이들이 전체 게놈상에 고르게 분포하고 있음을 확인할 수 있었다. 본 연구에서 관찰된 51개 유전자좌들이 게놈상에 고르게 분포하고 있다는 특성 때문에 게놈상의 특정부위에 편중되지 않은 유전정보를 얻을 수 있다는 장점이 있다. 즉, 기존의 RAPD 표지자들 중 상당수가 독립적인 연관군을 형성하는 것으로 알려져 있기 때문에 이들 연관군이 위치한 특정 부위의 DNA를 증폭하여 분석하는 RAPD 표지자에 비해서 I-SSR 표지자들이 유전 다양성을 추정하는데 더 유용한 표지자로 활용될 수 있을 것으로 생각되며, 이들 표지자들이 독립적인 진화의 과정을 겪을 것으로 기대되기 때문에 cladistic 방법에 의해 진화적 유연관계를 추정하는데 더 적합한 표지자로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.751-757
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• 2012

In order to develop specific genetic markers and determine the genetic diversity of Bangladeshi native cattle (Pabna, Red Chittagong) and exotic breeds (Sahiwal), randomly amplified polymorphic DNA (RAPD) analysis was performed using 12 primers. Genomic DNA was extracted from 20 cattle (local and exotic) blood samples and extracted DNA was observed by gel electrophoresis. Among the random primers three were matched and found to be polymorphic. Genetic relations between cattle's were determined by RAPD polymorphisms from a total of 66.67%. Statistical analysis of the data, estimating the genetic distances between cattle and sketching the cluster trees were estimated by using MEGA 5.05 software. Comparatively highest genetic distance (0.834) was found between RCC-82 and SL-623. The lowest genetic distance (0.031) was observed between M-1222 and M-5730. The genetic diversity of Red Chittagong and Sahiwal cattle was relatively higher for a prescribed breed. Adequate diversity in performance and adaptability can be exploited from the study results for actual improvement accruing to conservation and development of indigenous cattle resources.

• 한국자원식물학회지
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• 제22권2호
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• pp.152-158
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• 2009

갈대속 식물 두 종 달뿌리풀과 갈대의 종간 유연관계분석 및 수환경변화에 따른 종내 지역별 유연관계를 조사하기위해 RAPD 분석을 수행하였다. 총 9개의 oligoprimer를 이용한 효소중합반응에서 300 bp에서 1,900 bp 사이의 구간에서 142개의 유효한 polymorphic band를 확인하였다. Nei-Li의 genetic distance를 이용한 분석결과에 의하면, 비유사도지수가 달뿌리풀 개체군 종내에서는 0.012에서 0.061, 갈대 개체군 종내에서는 0.033에서 0.095의 낮은 상이성을 나타내어 종내 지역집단간에는 높은 유전적 유연관계를 보였으며, 달뿌리풀 개체군과 갈대 개체군 종간에서는 0.043에서 0.132의 상이성을 나타내어 달뿌리풀과 갈대 사이의 비유사도지수의 큰 차이는 보이지 않았으나 유전적으로 거리가 있는 것으로 나타났다. RAPD 결과 이 두 종 사이에 뚜렷한 차이를 구별할 수 있는 genetic marker를 확인하였고, UPGMA 유집분석에 의하면 두 종은 비슷한 수환경끼리 유연관계가 가까운 것으로 나타났고 다른 수환경끼리는 유연관계가 먼 것으로 나타났다. RAPD 분석법은 갈대속 두 종의 분류학적 혼동 해결과 동시에, 수환경변화에 따른 종내 지역별 유전적 유연관계 확인에 매우 유용한 방법인 것으로 나타났다.