Discrimination of Kalopanax pictus from its varieties and other ‘Kalopanacis Cortex’ plants by multiplex polymerase chain reaction (PCR)

도의정,Ningjie Yang,Myung-Guk Ji,이금산,주영승,이제현,Byong Seob Ko,오승은 한국유전학회 2011 Genes & Genomics Vol.33 No.6

‘Kalopanacis Cortex’ (KC) is an important medicinal material prescribed in Korea for the treatment of various ailments such as paralysis, arthritis, and rheumatism. In Korea, KC is defined as the dried stem bark of Kalopanax pictus. However, the stem barks of Zanthoxylum ailanthoides and Erythrina plants such as E. variegata have also been described as KC and prescribed for the same diseases. Since the pharmacological activities and contents of compounds isolated from the stem barks of these KC plants are obviously different, KC from K. pictus should be clearly discriminated from other stem bark in the best interest of public health. This study sequenced the internal transcribed spacers (ITS) of 52 samples of the KC or KC plants collected from Korea, China, and Vietnam. On the basis of different nucleotide sequences of specific ITS regions among the KC plants, the primer set KP F2/KC R1 was designed to amplify a 398-bp DNA marker for discriminating K. pictus from its varieties and from other KC plants. This primer set,along with the primer sets ZR F1/ZR R4, KP F1/EV R2, and KPF1/KC R1, was successfully amplified with the DNA markers of the Z. ailanthoides, E. variegata, and K. pictus taxa,respectively. The multiplex polymerase chain reaction method developed here not only discriminates the stem bark of K. pictus from stem barks of other KC plants but also identifies KC plants that supply KC in a single process.

營實과 그 위품의 유전자 감별

도의정,신상문,이금산 대한본초학회 2019 大韓本草學會誌 Vol.34 No.4

Objectives : Rosae Multiflorae Fructus is a traditional medicine derived from the fruit of Rosa multiflora Thunb. a member of the Rosaceae family. Even though it has a single origin, the possibility of adulterants has always existed. In fact, we had discovered suspicious commercial samples of Rosae Multiflorae Fructus, imported from China. Methods : To define the taxonomic origin of Rosae Multiflorae Fructus and its adulterants, DNA barcode analysis of the internal transcribed spacer, trnL-F intergenic spacer, and psbA-trnH sequences was carried out. These DNA barcode sequences from the correct origin of Rosae Multiflorae Fructus were analyzed and compared with those of other samples from genus Rosa used as medicinal herbs. Results : The analyses of the three DNA barcode sequences efficiently distinguished Rosae Multiflorae Fructus from six other species in genus Rosa and also separated each species used in this study. According to the DNA barcoding results, none of the suspicious commercial samples were Rosae Multiflorae Fructus. RMF09 was identified as Rosa acicularis, whereas RMF10 and RMF11 were identified as Rosa davurica and Rosa rugosa, respectively. These results corroborated the existence of adulterants of Rosae Multiflorae Fructus. Conclusions : Our research provides useful information that could be used as a criterion for distinguishing between Rosae Multiflorae Fructus and its adulterants. These results will help in the prevention of adulteration and also suggest effective methods for verifying the origin of commercial herbal medicines derived from genus Rosa.

川續斷과 韓續斷의 외부형태 감별

도의정,전신희,이금산 한약정보연구회 2015 한약정보연구회지 Vol.3 No.2

The authentication of Sokdan, one of the Korean traditional medicines, was performed with comparative study of morphological feature between Dipsaci Radix, roots of Dipsacus asper Wall. ex C.B. Clarke, and Phlomidis Radix, those of Phlomis umbrosa Turcz. In addition to a sensory observation, a stereoscope was used for more improve the accuracies. The surface of Dipsaci Rdix (DR) had more deeper furrows than it of Phlomidis Radix (PR). And more, the surface patterns of PR were more regular and fine than those of DR. In a section, DR did not have a clear distinction between nearby xylem, but PR did. In a distal section of DR, the color of cortex was dark-brown or dark- green and xylem was lemon yellow. On the other hand, PR was generally lemon yellow. It was discriminable for two herbs by observing pattern of furrows and colors of section.

Differentiating Coptis chinensis from Coptis japonica and other Coptis species used in Coptidis Rhizoma based on partial trnL-F intergenic spacer sequences

도의정,오승은,이미영,고병섭 한국유전학회 2014 Genes & Genomics Vol.36 No.3

Dried rhizomes of Coptis species are utilized as‘‘Coptidis Rhizoma’’ (CR), an important herbal medicinalmaterial in traditional Chinese medicine. Almost all CRstraded in the Korean herbal medicine market originate fromCoptis chinensis (‘‘Chun Hwang-Lyun’’ in Korean medicalterminology). Other minor CRs originate from Coptisjaponica (‘‘Il Hwang-Lyun’’). Although there is an obviousdiscrepancy in the price of traded CRs in the herbal marketdepending on the Coptis species, CRs originating from C. chinensis and C. japonica are often confused. Furthermore,the CR traded as ‘‘Chun Hwang-Lyun’’ is occasionallymixed with rhizomes of Coptis deltoidea and/or Coptisomeiensis. Therefore, we sought to discriminate C. chinensisfrom C. japonica, as well as C. deltoidea and C. omeiensis, by using nucleotide sequence differences in thepartial trnL-F intergenic spacer. We developed an efficientreal-time polymerase chain reaction (PCR)-based discriminationassay to separate samples of C. chinensis fromthose of C. japonica without the need to separate the DNAmarkers by using gel electrophoresis. In addition, wedeveloped a multiplex PCR method with which we wereable to discriminate samples of C. chinensis from those ofC. deltoidea and C. omeiensis by amplifying the 153-bpDNA marker in C. chinensis in a single PCR process.

천연보존제를 위한 특정 한방처방의 추출용매에 따른 항균활성 비교

도의정,백승호,이금산 한약정보연구회 2015 한약정보연구회지 Vol.3 No.1

This study was conducted to investigate the new candidates of natural preservatives, modified by based on the Korean medical prescription. We created 4 candidate extracts with different solvents. The crude water extract JBTP01 and JBTP02 showed weak antibacterial activities. Otherwise, 70% EtOH extract JBTP03 and MeOH extract JBTP04 showed wide antibacterial spectrum. Both were shown strong antibacterial activity to E. coli. JBTP03 was also effected bacteria C. albicans. Compared the antibacterial activities with methylparaben, commonly used syntetic antiseptic, and 3 natural preservatives in market. JBTP03 and JBTP04 showed similar modality of antibacterial activity spectrum. In other words, JBTP03 and JBTP04 had strong possibility to be new natural preservatives. Further study need to be done to use JBTP03 and JBTP04 for natural preservatives in cosmetics.

밀몽화(密蒙花)와 결향화(結香花)의 외부형태감별

도의정,이금산,정승일 한약정보연구회 2013 한약정보연구회지 Vol.1 No.-

Objectives: For the authentication of the Milmong-wha, one of korean herbal medicines, it was made a comparative study between Buddlejae Flos (BF), flower bud or inflorescence of Buddleja officinalis Maxim., and Edgeworthiae Flos (EF), those of Edgeworthia chrysantha Lindl. Methods: In addition to a sensory observation, a stereoscope was used for more accuracies. Results: BF was yellowish-brown and irregularly truncated conical, whereas EF was stramineous and regularly hemispherical. The flower bud of BF was densely stellate tomentose with some glandular hairs, while the that of EF was densely white silkily hairy. Conclusions: The type of hairs and general appearance are discriminable criteria between two herbs, BF and EF.

DNA Barcode를 이용한 Cinnamon (Cinnamomum spp.) 감별 및 국내 유통 현황 모니터링

도의정,오승은,이금산 한국약용작물학회 2015 한국약용작물학술대회 발표집 Vol.2015 No.10

백수오(白首烏)와 이엽우피소(耳葉牛皮消)의 내부형태 감별기준

도의정 ( Eui-jeong Doh ),김정훈 ( Jung-hoon Kim ),최고야 ( Goya Choi ),이승호 ( Seung-ho Lee ),송호준 ( Ho-joon Song ),주영승 ( Young-sung Ju ),이금산 ( Guem-san Lee ) 대한본초학회 2015 大韓本草學會誌 Vol.30 No.4

Objectives: The root of Cyanchum wifordii (CW) had been used as herbal medicine, ``Baeksuo``, in Korea. However, the root of Cynanchum auriculatum (CA) had also been mis-used as ``Baeksuo`` in herbal markets, due to its morphological similarity with CW. This study aimed to compare discriminative features and establish the identification-keys between two herbal medicines using microscopic methods. Methods: Microscopic preparations including fixation, rapid dehydration, paraffin infiltration, paraffin embedding, sectioning, mounting, and staining, were performed and the microscopic features between CW and CA samples were compared. Results: The starch grain of parenchymal cells, the amounts of calcium oxalate crystals, cell sizes, and the shape of phloem were not different between CW and CA samples. However, the clusters of stone cells which were conglomerated at outer cortex were apparently observed in CA sample (4~8 layers of stone cells), but not in CW sample (2~4 layers of stone cells, or not). Xylem of CW consisted of several vessels and fiber bundles which were formed as linear shapes and discontinuous radial lines, while that of CA, in which radial lines was not observed, were shaped as a wedge and scattered near cambium with comparatively few fiber bundles enclosed vessels. Especially, in the xylem of CA, fiber bundles located towards cambium were relatively more developed than those located outward cambium. Conclusions: The identification-keys established in this study would be helpful to discriminate microscopic features between CW and CA.

Multiplex PCR을 이용한 4 종류 목향(木香)의 감별

도의정 ( Eui Jeong Doh ),이금산 ( Guem San Lee ),주영승 ( Young Sung Ju ),오승은 ( Seung Eun Oh ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.3

Objectives : Aucklandiae Radix (Muxiang) one of important herbal medicines in oriental medicine, is defined as the dried root of Aucklandia lappa (Asteraceae). Owing to the similarities in the morphology and name, Inulae Radix (Tu-Muxiang) and Vladimiriae Radix (Chuan-Muxiang) as well as Aristolochiae Radix (Qing-Muxiang) originated from other medicinal plants are often used as substitutes and/or adulterants of Aucklandiae Radix. Therefore, a reliable authentication of these herbal medicines is necessarily for the public health and prevention of misuse. Methods : 32 samples of medicinal plants supplying Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix were collected in Korea and China. The ITS (Internal transcribed spacer) nucleotide sequences of samples were determined. The PCR primers to amply DNA marker of each herbal medicine were designed basing on the specific ITS regions showing differences in the sequences among medicinal plants. Results : Primer set Al R/IS F designed in this work amplified 220 bp PCR product only in samples of Aucklandiae Radix. In contrast, primer set Ih F/IS R, Vs R/IS F, and AcR F1/Ac R amplified 250 bp product, 356 bp prouct, and 516 bp product respectively to identify Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. Conclusions : The primers designed basing on the nucleotide sequences of ITS regions appearing differenced in the sequences among medicinal plants amplified the DNA markers for the identification of Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. These herbal medicines were more efficiently identified by multiplex PCR method using all primers in a single PCR process.

대한민국약전에 수재된 식물성 한약재의 학명에 대한 재고

도의정 ( Eui Jeong Doh ),이금산 ( Guem San Lee ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.3

Objectives: A precise and simple system of nomenclature was required to avoid error, ambiguity or confusion. Although medicinal plants must be produced or distributed based on a pharmacopoeia described origin including scientific name, the Korean Pharmacopoeia tenth edition (KP 10) had many names against the nomenclature. Therefore, this study aimed at searching correct scientific names for 241 plants in KP10. Methods: Authoritative databases - The Plant List, International Plant Name Index, YList, Tropicos, eFloras, World Checklist of Selected Plant Families, The Global Compositae Checklist, The International Legume Database and Information Service, et al. - and previously performed researches, floras were cross-checked. Results: The arrangement of this list was designed for four cases, errors including illegitimate, nomenclatural synonyms, recommended names and decision reserved names. Consideration about the scientific names produced nine correct names for ten misspellings and illegitimate, and thirty-six correct names for forty-one nomenclatural synonyms. These results should be reflected in the next of KP10. Separately. ten recommended names were also suggested for taxonomic synonyms which had been used indiscriminately due to diverse taxonomic opinions, In addition to those, decision reserved names were suggested for thirteen species which had been corridor of uncertainty, Then again, there was need to study about authorship, because KP 10 did not keep recommendations for author citations. Conclusions : Correction of scientific names for some medicinal plants which violated the International Code of Nomenclature would be useful to improve the accuracy of a Pharmacopoeia as the criterional materials.