한국산 나문재속 식물에 대한 계통학적 유연관계를 밝히고, 분자계통학적 연구를 통해 나문재속 종간 유연관계를 확인할 수 있는 분자마커를 찾아내기 위해 연구를 수행하였다. 핵 리보솜 DNA ITS와 엽록체 DNA matK, psbA-trnH 그리고 trnL-trnF를 분자마커로 사용하였다. ITS 영역은 칠면초와 해홍나물 그리고 해홍나물과 방석나물을 구분하지 못하였다. psbA-trnH와 trnL-trnF 영역의 염기서열은 칠면초와 방석나물을 구분하지 못하였다. 그러나 4종의 분자마커 영역을 조합하여 분석한 결과 나문재속 식물 5종이 각각 독립적인 계통을 형성하는 것을 확인하였다. 따라서 나문재속 계통관계 분석을 위해서 여러 개의 분자마커 조합이 유용할 것으로 판단된다. 나문재속 내 분류군 간의 계통관계를 명확히 밝히기 위해 차후에 좀 더 많은 생태학적, 형태학적 자료를 조사해야 할 것으로 보인다.

다양한 환경에서 수집한 국내 민들레속 유전자원 수집종의 엽록체 DNA 영역(trnL-trnF와 rps16-trnK) 염기서열을 이용하여 종내․ 간 변이 및 배수성을 구명하여 유전자원 육성의 기초 자료룰 제공하고자 수행하였다. 민들레속 유전자원의 배수성은 털민들레, 서양민들레, 붉은씨서양민들레가 3배체이고, 흰 민들레와 흰노랑민들레는 4배체였다. 염기서열의 길이는 trnLtrnF 영역에서 자생종류인 털민들레, 흰민들레, 흰노랑민들레 가 931 bp에서 935 bp, 서양민들레는 910 bp, 붉은씨서양민들레 는 975 bp로 종간 차이를 나타내었고, 종 특이적 염기서열 88개, 자생종 및 귀화종 특이적 염기서열 41개가 검출되었다. rps16- trnK 영역은 털민들레 882∼883 bp, 흰민들레 875∼881 bp, 흰 노랑민들레는 878∼883 bp 서양민들레 874∼876 bp, 붉은씨서 양민들레는 847∼848 bp로 37개 종특이적 염기서열이 검출되 었다. 염기서열의 유사도는 trnL-trnF 영역에서 0.860∼1.000 사이로 평균 0.949이며, rps16-trnK 영역의 유사도는 0.919∼ 1.000 사이로 평균 0.967이었다. 염기서열을 바탕으로 유연관계를 분석한 결과, trnL-trnF 영역은 크게 자생종류와 귀화종 류로 구분되었으며, 서양민들레와 붉은씨서양민들레는 같은 종 간에 유집되었고, 자생종류는 분리되지 않았으며, rps16-trnK 4개 그룹과 유집되지 않은 5개체로 나뉘었다. 흰노랑민들레는 두 영역 모두 흰민들레와 동일 계통군을 형성하였고, 염기서열 상 두 종간 뚜렷한 차이가 없었다. 유연관계에서 모두 독립적으로 존재한 흰민들레 No. 10 (조계산)과 털민들레 1번(광양)은 민들레 유전자원 육성소재로 활용이 기대된다.

Background : In the herbal medicine market, Angelica gigas, Angelica sinensis, and Angelica acutiloba are all called "Danggui" and used confusingly. We aimed to assess the genetic diversity and relationships among 14 Angelica species collected from different global seed companies. Toward this aim we developed DNA markers to differentiate the Angelica species. Methods and Results : A total of 14 Angelica species, A. gigas, A. acutiloba, A. sinensis, A. pachycarpa, A. hendersonii, A. arguta, A. keiskei, A. atropurpurea, A. dahurica, A. genuflexa, A. tenuissima, A. archangelica, A. taiwaniana, and A. hispanica were collected. The genetic diversity of all 14 species was analyzed by using five chloroplast DNA-based simple sequence repeat (SSR) markers and employing the DNA fragment analysis method. Each primer amplified 3 - 12 bands, with an average of 6.6 bands. Based on the genetic diversity analysis, these species were classified into specific species groups. The cluster dendrogram showed that the similarity coefficients ranged from 0.77 to 1.00. Conclusions : These findings could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Angelica species. The analysis of polymorphic SSRs could provide an important experimental tool for examining a range of issues in plant genetics.

Background: In the herbal medicine market, Angelica gigas, Angelica sinensis, and Angelica acutiloba are all called "Danggui" and used confusingly. We aimed to assess the genetic diversity and relationships among 14 Angelica species collected from different global seed companies. Toward this aim we developed DNA markers to differentiate the Angelica species. Methods and Results: A total of 14 Angelica species, A. gigas, A. acutiloba, A. sinensis, A. pachycarpa, A. hendersonii, A. arguta, A. keiskei, A. atropurpurea, A. dahurica, A. genuflexa, A. tenuissima, A. archangelica, A. taiwaniana, and A. hispanica were collected. The genetic diversity of all 14 species was analyzed by using five chloroplast DNA-based simple sequence repeat (SSR) markers and employing the DNA fragment analysis method. Each primer amplified 3 - 12 bands, with an average of 6.6 bands. Based on the genetic diversity analysis, these species were classified into specific species groups. The cluster dendrogram showed that the similarity coefficients ranged from 0.77 to 1.00. Conclusions: These findings could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Angelica species. The analysis of polymorphic SSRs could provide an important experimental tool for examining a range of issues in plant genetics.

Background : Plants belonging to 5 species of the genus Eleutherococcus are currently distributed in the Korean peninsula. The traditional medicine ‘Ogapi’, derived from Eleutherococcus sessiliflorus and other related species, and ‘Gasiogapi’, derived from Eleutherococcus senticosus, are frequently mixed up and marketed. Therefore, accurated identification of their origins in urgently required. Methods and Results : Candidate genes from nuclear ribosomal DNA (nrDNA) and chloroplast DNA (cpDNA) of Eleutherococcus plants were analyzed. Whereas the nrDNA-internal transcribed spacer (ITS) regions were useful in elucidating the phylogenetic relationships among the plants, the cpDNA regions were not as effective. Therefore, a combined analysis with nrDNA-ITS was performed. Various combinations of nrDNA and matK were effective for discriminating among the plants. However, the matK and rpoC1 combination was ineffective for discriminating among some species. Based on these results, it was found that OG1, OG4, OG5, OG7, GS1, GS2, and GS3 were derived from E. sessiliflorus. In particular, it was confirmed that GS1, GS2, and GS3 were not derived from E. senticosus. However, more samples need to be analyzed because identification of the origins of OG2, OG3, OG6 and GS4 was not possible. Conclusion : The ITS2, ITS5a, and matK combination was the most effective in identifying the phylogenetic relationship among Eleutherococcus plants and traditional medicines based on Eleutherococcus.

Polygonatum is a genus placed in the family Liliaceae, distributed throughout the Northern Hemisphere and 16 of the species are grown naturally in Korea. In oriental medicine, the rhizomes of Polygonatum have been used as two different medicines, Okjuk (Polygonati odorati Rhizoma) and Hwangjeong (Polygonati Rhizoma). However, it is difficult to identify the morphological and chemical differences between the medicinal groups and thus easy to confuse the one with the other. Therefore, a clear classification standard needs to be established so as to be able to discriminate between them. In the study, the morphological characteristics of the plants, Polygonatum spp., were examined. Then, the differences in SNPs among the DNA sequences of 7 of the Polygonatum spp. and 1 of the Disporum spp. were analyzed by DNA barcoding with rpoC1, rpoB2, matK, and psbA-trnH of the cpDNA region. In the results, three regions, rpoC1, rpoB2, and matK were useful for discriminating the species, P. stenophyllum and P. sibiricum. Furthermore, it was possible to discriminate the individual germplasm within the species by using the combination of the results obtained from rpoB2, rpoC1, and matK.

We performed phylogenetic analyses of a total of 21 acessions covering 5 species in the Korean Trigonotis and one outgroup species using nuclear ribosomal ITS and chloroplast rbcL, matK, ndhF sequences. Outgroup were chosen from the closely related genus Lithospermum zollingeri. Both parsimony and Bayesian Inference methods were used to reconstruct the evolutionary history of the group. The evidence collected indicated that phylogenetic relationships among Korean Trigonotis species are unresolved based on nuclear marker (ITS), as the same as based on separated chloroplast sequences. While the phylogenetic relationships of Korean Trigonotis species almost clearly were resolved in combined chloroplast sequences. Thus, the members of Trigonotis coreana can be distinguished to the members of Trigonotis peduncularis in combined cpDNA sequences and Trigonotis nakaii was treated as a synonymed to Trigonotis radicans var. sericea. In addition, the MP and BI analysis showed Trigonotis icumae as sister of the remained Korean Trigonotis species based on combined molecular markers (BI: PP = 1).

Next generation sequencing (NGS) approaches can also be useful tool for characterization of organelle genomes. We generated chloroplast (CP) genome sequences of two Korean ginseng cultivars, Chunpoong and Yunpoong, based on reference-guided assembly using whole genome NGS data. We used 0.5x of P. ginseng genome NGS reads to assemble CP genome. Of the NGS reads used, about 6% were mapped to the reference CP genome with mean coverage of 94x due to high copy number of CP genome in plant cell. CP genomes of the two cultivars were predicted to be 156,248 bp and 156,355 bp in length and showed about 0.1% differences at nucleotide level, compared to reference CP genome sequenced from P. ginseng (Acc.no. NC_006290), whereas difference between CP genomes of the two cultivars is very rare. In this study, we developed the molecular marker to perform taxon identification and also to elucidate phylogenetic relationship among Korean ginseng cultivars. Now, we are analyzing the CP genomes of other P. ginseng cultivars together with other Panax species including American ginseng and Panax related species.

There is a considerable difference in morphological traits between Bokbunja cultivated in Korea (KCB) and Korea native Rubus coreanus, contrary to the conviction that the cultivated Bokbunja is the domestication of R. coreanus. To infer the phylogenetic relationship of KCB with other Rubus species, we compared the chloroplast DNA spacers of KCB with those of several Rubus species including black raspberry, R. occidentalis. The three chloroplast DNA spacers, atpB~rbcL, trnL~trnF, and trnT~trnL, were amplified using the specific primer pairs and converted to Single Strand Conformational Polymorphism (SSCP) markers. The SSCP makers of the chloroplast DNA spacers showed a considerable variation both within and among Rubus species. In the phylogenetic tree generated by the SSCP markers, KCB accessions were located in the same clade with R. occidentalis, but R. coreanus accessions in the different clade. Also, in the phylogenetic tree by the nucleotide sequences of the chloroplast DNA spacer trnL~trnF, KCB located in the same clade with R. occidentalis but not with R. coreanus. These results suggest that the three KCB accessions share higher similarity with R. occidentalis than with R. coreanus in the three chloroplast DNA spacers.

지모 (Anemarrhena asphodeloides)는 탁월한 해열작용과 진정작용을 갖는 한약재로 한국, 중국, 일본에서 널리 이용되어 왔다. 본 연구에서는 먼저 국내 연구소에서 형태학적 분류 결과 지모로 확인된 3종의 식물체를 수집하여 엽록체 DNA의 trnL-F 염기서열을 분석하였다. 분석 결과, 국내 연구기관에서 보관중인 지모 식물체들이 모두 동일한 trnL-F의 염기서열을 보여서, 형태학적 분류와 계통유전학적 분류가 동일함을 확인하였다. 최초로 얻어진 지모 trnL-F 염기서열은 NCBI database에 등록하였다. 다음으로 국내 한약재 시장과 중국 한약재 시장에서 유통 중인 지모 한약재를 다량 구입하여 trnL-F의 염기서열을 분석하였다. 그 결과, 유통 중인 지모 한약재들이 모두 기원식물과 동일한 TrnL-F의 염기서열을 보여서 지모 약재의 경우 진품이 유통되고 있음을 알 수 있었다. TrnL-F의 염기서열로 계통수를 작성한 결과 지모는 아스파라거스목 (Asparagales), 용설란과 (agavaceae)에 속한 것으로 보여 졌다. 엽록체 rbcL 유전자 염기서열로 얻은 계통수와 비교한 결과 trnL-F 계통수와 rbcL 계통수가 비슷한 결과를 보여주어서 분자유전학적 분류에 두 유전자가 상호보완적으로 이용될 수 있음을 확인하였다.