본 연구는 국내 유통 방풍(S. divaricata, P. japonicum, G. littoralis) 한약재의 원산지 판별을 위해 수행 되었다. 이를 위 해, 형태적 특징비교 뿐만 아니라 엽록체(nrDNA-ITS2) 및 핵 DNA 바코드 유전자(cpDNA-matK, psbA-trnH, rpoB2와 rpoC1)를 이용한 염기서열 분석을 수행하였다. 방풍류 3종의 형태적 특징을 비교한 결과, 잎 모양과 거치의 형태에서 가장 큰 차이를 보인 반면, 건조약재의 경우 육안으로 구별하기에 어려움이 있었다. DNA 수준에서의 차이를 비교하기 위해 DNA 바코드 후보 유전자들을 이용한 방풍류 3종의 식물자원 에 대한 염기서열 분석을 수행한 후 판별 마커 개발 가능성이 있는 ITS2와 matK, psbA-trnH 세 primer를 선발하였다. 이 를 국내 유통 한약재에 적용한 경우 방풍은 S. divaricata와, 식방풍은 P. japonicum과, 해방풍은 G. littoralis와 동일한 것 으로 확인되었다. 중국산 방풍은 S. divaricata와 동일종으로 바르게 표기되어 유통되나, 국산 방풍은 P. japonicum과 동일 종으로 식방풍(갯기름나물)의 뿌리를 건조시켜 방풍으로 혼용 유통됨을 확인하였다. ITS2 구간의 염기서열 분석 결과, 식방 풍의 경우 두 그룹으로 나눠져 동일 종 내에 유전적 변이가 있음이 확인되었다. 따라서 본 연구결과 방풍류 3종 판별을 위 해 nrDNA-ITS2와 cpDNA-matK, psbA-trnH의 사용이 유용 할 것이며, 차후 방풍류 한약재의 판별을 위한 마커 개발 연 구의 기초 자료로서 활용될 수 있을 것으로 사료된다.

Background : Compositae is one of the largest plant families which has high probability of diversity and mutation. Though the various researches about the Compositae are ongoing, it is incomplete and need to be conducted the molecular genetic researches to back up the previous studies. This research was performed to identify the genetic divergence of Compositae plants based on the DNA barcoding for cpDNA-matK and rbcL regions. Methods and Results : For this studies, the genetic sequence analysis (SNP/InDel) and phylogenetic analysis were conducted by using Neighbor-Joining algorithm as targeting the nineteen specimens from 7 species which received a IT number along with NCBI Genbank database (http://ncbi.nlm.nih.gov) sequences. The result of matK sequence analysis, 68 SNP and 2 InDel regions (at nt 527-538bp and 695-706bp positions) were confirmed. Also 10 SNPs were found in rbcL region. The genetic divergence showed 0.000-0.059% in matK regions, and the mean was 0.024%. The highest distance were observed between Ligualria fischeri and the group composed with Aster tataricus and Solidago virgauria (2 and 3). The sequence divergence for rbcL regions showed 0.000-0.018%, and the mean was 0.005%. The highest sequence distance in rbcL region were observed between L. fischeri group and S. virgauria (HE574593). In result of phylogenetic analysis in matK region, the most species formed independent clade. A. tataricus in Aster genus and two samples of S. virguaria in Solidago genus were formed one same clade. S. virguaria(1) and A. spathulifolius(2) has been separated into independently for the plants belonging to same genus, respectively. A. spathulifolius showed differences with NCBI data. The rbcL formed one same clade except L. fischeri and Synurus deltoides. Conclusion : This study indicates that matK is more valuable than rbcL for the distinction among the species of Compositae. This results are expected to be used for the establishment of the classification system of Compositae as well as for the studies in the development of an authentication marker.

Background : Ixeris dentata is the perennial herbaceous plant in the Ixeris genera within the Compositae family. The whole plant has been used traditionally as herbal medicine. It is widely distributed in South Korea and the genetic difference among the plants harvested from different regions may differ due to the disparity in cultivation climate. Therefore, this research was performed to discriminate the I. dentata that are collected from four locations in South Korea based on sequence analysis of nrDNA-ITS region. Methods and Results : Genomic DNA was extracted from I. dentata obtained from Goesan, Dangjin, Yangpyeong, and Chuncheon. I. stonolifera was included for a comparison of genetic distance with I. dentata. PCR amplification was performed by using an universal barcode nrDNA-ITS primer for DNA barcoding. After sequencing, the data was aligned using ClustalW multiple alignment tool in BioEdit version 7.2.5 software. SNP and phylogenetic analysis were conducted with the MEGA7 program. Phylogenetic analysis was presented with Neighbor-joining method using the K2P model. Statistical significance are evaluated using bootstrap (1,000 replicates). PCR products were amplified with about 800bp length for the ITS1-4 sequences of all samples. Nineteen SNPs were detected within a 578bp fragment of the aligned sequences. The mean GC content was 51.88% for ITS1-4 sequences of them. The interspecific genetic distance between I. dentata and I. stonolifera was 0.029%. The highest region-specific distance was confirmed to 0.010% between the plants from Dangjin with Goesan and Chuncheon group. Meanwhile, the mean intraspecific distance among the plants from Yangpyeong was 0.002%. Conclusion : In the phylogenetic analysis, the plants from Goesan and Chuncheon were placed within the same clade, while the plants from Dangjin formed independent clade. On the other hand, the plants from Yangpyeong were not grouped into one clade followed by the intraspecies variation. In conclusion, the data from the research will be useful for the regional identification of Dangjin, even if it is required to perform some additional researches.

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.

This study was conducted to evaluate the allelopathic effect of aqueous extract of Ganghwa domestic mugwort (Artemisia spp.) on vegetables and its related allelochemicals. When the receptor vegetables, such as Chinese cabbage, lettuce, and red radish, were treated with aqueous extract obtained from Sajabalssuk (A. sp*I), Ssajuarissuk (A. sp*II) or Ssajarissuk (A. sp*III), their germination rate, leaf number, plant height, and root length were restricted with increasing concentration of aqueous extract. Allelopathic effect was the highest in radish, than lettuce and Chinese cabbage in order. The growth of topplant were more inhibited then root growth observing in restriction of plant height, root length, and chlorophyll contents. The plant height, the root length of red radish were 53.3 and 61.2% and their fresh weights were 19.8 and 26.4% compared to those of controls, respectively. A. sp*III showed the highest allelopathic effect among the donor plants. In HPLC analysis, 7 phenol compounds were identified in A. sp*I and A. sp*II, and, in A. sp*III, and hydroxybenzoic acid and phenylacetic acid were further identified as allelochemicals. It is considered that their plant growths were variously inhibited by the amounts and types of allelochemicals in aqueous extracts. To increase the productivity of farm land after cultivation of mugwort, these results can be useful to select the following field crops.

This study was conducted to identify allelopathic effect of Ganghwa domestic Artemisia spp., named Sajabalssuk and Ssajuarissuk, for various receptor plants including clover (Trifolium repens L.), alfalfa (Medicago sativa L.), lawn grass (Zoysia japonica Steud.), dandelion (Taraxacum platycarpum Dahlst.), and dahurianpatrinia (Patrinia scabiosaefolia Fisch. ex Trevir). Receptor plants were treated with the aqueous and essential oil extract of Artemisia plants. In consequence, their allelopathic effects were evaluated by measuring seed germination rates, seedling growth, and dry weights of the receptor plants. The seed germination and seedling growth of the receptor plants were inhibited by all treatments of both aqueous and essential oil extracts of the Artemisia plants, and, in addition, the inhibitory effects were increased according to the higher concentration. Among the donor plants, A. sp.*Ⅲ showed most effective allelopathic effect. Comparing the alleopathic effect among the receptor plants, seed germination was most inhibited in lawn grass while inhibitory effect of seedling growth was comparatively higher in dandelion. Although inhibitory effects were comparatively lower, the allelopathic effects of Artemisia plants were identified in clover and alfalfa since the seedling growth of these plants were inhibited more than 70%. Thus, in result, Ganghwa domestic Artemisia spp. could be possibly used for weed control since natural products of the plants showed inhibitory effects on seed germination and seedling growth of various receptor plants.

본 연구는 최근 한방자원, 사료자원, 바이오에너지 자원 등 다양하게 이용되는 국내 자생 억새(Miscanthus sinensis)의 대량생산 및 신품종 개발을 위한 조직배양체계 확립을 위해 수행되었다. 이를 위해 억새 완숙종자로부터의 캘러스 유도와 재분화를 위한 식물생장조절제의 적정농도를 규명하였다. 억새의 성숙종자유래 배발생 캘러스 유도를 위해 2,4-D, IBA, NAA를 1~10 mg·L-1의 농도로 단용 처리한 결과, 5 mg·L-1 2,4-D 처리에서 가장 높은 85.3%의 캘러스 유도율과 캘러스의 증식을 보였으며 조직배양 과정 중 갈변화율도 가장 낮았다. 또한, 캘러스의 재분화를 위해 옥신인 NAA와 Kinetin, 2-iP, 또는 BAP 등의 사이토키닌을 혼용 처리한 결과, 각각 19.0%~59.0%, 23.0%~67.3%, 14.7%~83.7%의 재분화율을 보여 NAA와 BAP의 혼용 처리구가 NAA와 Kinetin 또는 2-iP와 혼용 처리구보다 식물체 재분화에 효과적이었다. 특히 3 mg·L-1 NAA와 5 mg·L-1 BAP 혼용 처리된 배지에서의 재분화율이 83.7%로 가장 높게 나타났으며, 캘러스 당 재분화 식물체 개수도 5.5개로 동일농도의 2-iP 또는 Kinetin 혼용 처리 시 2.1 및 2.0개보다 많았다. 본 연구결과 억새 성숙 종자로부터의 배발생 캘러스 유도에는 5 mg·L-1 2,4-D가 그리고 캘러스의 재분화에는 3 mg·L-1 NAA와 5 mg·L-1 BAP 혼용 처리가 가장 효율적이었다. 본 연구를 통해 확립된 조직배양체계는 억새의 대량생산 및 신품종 개발에 유용할 것으로 판단된다.