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Development of species specific RAPD markers using populus cpDNA
Jae Soon Lee(李載順)Eun Woon Noh(盧銀雲),Suk Sung Jang(張錫成),Suk Koo Lee(李錫求),Eui Rae Noh(盧義來),Don Koo Lee(李敦求) 한국육종학회 1994 한국육종학회지 Vol.26 No.4
Chloroplast DNAs(cpDNA) from eight different species of Populus(P. alba, P. glandulosa, P. alba×P. glandulosa, P. davidiana, P. maximowiczii, P. nigra×P. maximowiczii, P. nigra, and P. Koreana×P. nigra), Salix pseudo-lasiogyne and Nicotiana tabacum were compared by PCR amplification. The cpDNAs were amplified at relatively low annealing temperature so that many bands could be observed. Amplification of a spacer between 16S γRNA gene and 23S γRNA gene located in inverted repeats in cpDNA resulted in a major band in size of 2.3 Kb and several minor bands of various sizes. A band specific to Populus was found when the amplification was done at lower annealing temperature. Amplification with primers deduced from γpoC1 and γpoC2 gene produced the major target band as well as several additional (minor) bands that could be used for the differentiation of each Populus species. The minor bands appeared repeatedly even when different thermocycles were employed. With the amplification profile, it was possible to differentiate Populus species.
Genetic relationship among acer species revealed by RAPD and PCR-RFLP
Jae Woo Hwang(黃在禹),Eun Woon Noh(盧銀雲),Jae Soon Lee(李載順),Sook Kyung Park(朴淑京) 한국육종학회 1996 한국육종학회지 Vol.28 No.4
Ten different species of Acer including A. triflorum, A. saccharinum, A. pseudo-sieboldianum, A paimatum var. sanguineum, A. mono, A. ruburm, A. negundo, A. palmatum, A ginnala, and A. tegmentosum were compared with regard to their DNA variation based on RAPD profiles and PCR-RFLP. For RAPD study, only five species(A. triflorum, A. saccharinum, A pseudo-sieboldianum, A. palmatum var. sanguineum, and A. mono) consisting of more than 5 individuals were compared. For PCR-RFLP, 6 species that produced the target bands with the sequence specific primers were considered. Forty four loci were generated by RAPD with 4 arbitrary primers. Similarity between individuals was estimated by calculating the frequency of shared bands out of total bands. Cluster analysis based on the similarity produced two groups. While the first group was exclusively composed of A. pseudo-sieboldianum, the second comprised the remaining 4 species. When the primers specific to the gene sequences rpl2 and psbA in chloroplast DNA were used, A pseudo-sieboldianum again appeared to have different amplification pattern from the rest of Acerspecies examined. Analysis of the amplified psbA-rpl2 region(1.7 kb) by PCR-RFLP revealed that the species in the genus could be readily differentiated from each other by the fragmentation profile. We found that although RAPD worked on Acer species, it costed too much time and labor for the optimization of PCR conditions. On the other hand, the PCR-RFLP technique is very simple and straight forwarded to get reproducible results since it uses very stringent reaction conditions.
Isolation of nod F gene from Frankia spp. symbiotic with alnus species
Min Il Choi(崔敏逸),Don Koo Lee(李敦求),Eun Woon Noh(盧銀雲),Jae Soon Lee(李載順) 한국육종학회 1995 한국육종학회지 Vol.27 No.1
Frankia spp. are actinomycetes known to have symbiotic relationship with many actinorhizal woody species including Alnus and Eleagnus spp. Although they are known to infect those woody species by the similar mechanisms which Rhizobium and Bradirhizobium colonize legume species, most of the mechanisms concerning the nodule formation have remained unknown. In this study, we examined whether Frankia spp. have the same nod F gene as that of Rhizobium by amplification of the gene with the primers deduced from Rhizobium sequence. The amplification product and subsequent southern hybridization with nod F gene from Rhizobium meliloti confirmed that Frankia also carries the gene(or DNA segment) comparable to nod F gene in Rhizobium.