J Plant Biotechnol (2016) 43:151 156 DOI:http://dx.doi.org/10.5010/JPB.2016.43.2.151 ISSN 1229-2818 (Print) ISSN 2384-1397 (Online) Review Angelica 속식물의종판별을위한연구현황및전망 길진수 박상익 이이 김호방 김성철 김옥태 차선우 정찬식 엄유리 Current status and prospects of the authentication of Angelica species Jinsu Gil Sang ik Park Yi Lee Ho Bang Kim Seong-Cheol Kim Ok-Tae Kim Seon-Woo Cha Chan Sik Jung Yurry Um Received: 19 April 2016 / Revised: 11 May 2016 / Accepted: 11 May 2016 c Korean Society for Plant Biotechnology Abstract Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, natural cosmetics, and pharmaceutical industries. However, names are different from each country and their phylogenetic origins are not clear. These lead consumers to be confused. In particular, when they are morphologically similar and distributed as dried roots, it is extremely difficult to differentiate their origins even by specialists. Recently, molecular markers have been extensively applied to identify the origin of many crops. In this review, we tried to overview the current research achievements for the development of suitable origin identification regarding to the differentiation of Angelica species. Furthermore, more advanced techniques including amplification genome based marker analyses are also discussed for their practical applications in the authentication of particular medicinal plant in Angelica species. Keywords Angelica, Molecular marker, Origin identification S.-C. Kim O.-T. Kim S.-W. Cha C. S. Jung Y. Um ( ) 농촌진흥청국립원예특작과학원인삼특작부약용작물과 (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, Rural Development Administration, Eumseong 27709, Korea) e-mail: urspower@korea.kr J. Gil S.I. Park Y. Lee 충북대학교농업생명환경대학특용식물학과 (Department of Industrial Plant Science and Technology, Chungbuk National University, Cheongju 28644, Korea) H. B. Kim 바이오메딕생명과학연구소 (Life Sciences Research Institute, Biomedic Co., Ltd., Bucheon 14548, Korea) 서론 당귀는산형과에속하는 Angelica 속다년생초본으로써우리나라에서자생하는참당귀 (Angelica gigas), 중국에서자생하는중국당귀 (A. sinensis), 일본에서자생하는일당귀 (A. acutiloba) 가대표적이다. 우리나라에서많이재배되고있는참당귀와일당귀는한약재와쌈채소로각각이용되고있으며 2015 년농림식품부통계자료 (MAFRA, 2015) 에따르면참당귀는 1,345 톤, 일당귀는 274 톤이생산되고있지만수요량에비해생산량이적기때문에참당귀의경우국내생산량과비슷한수준의양을중국으로부터수입하고있는실정이다. 최근에우리나라는세계여러나라와 FTA 를체결하면서국내산농산물과외국산농산물에대한원산지관리, 국내에서개발된품종에대한지적재산권보호및우수품종에대한품질관리를위해서품종판별기술이기초기반기술로의가치를인정받고있다. 하지만수입되는대부분의한약재는가공처리후절편이나타블렛형태로판매되는경우가많아형태에의한품종구분은사실상불가능한실정으로 DNA 수준에서품종판별시스템을구축하는것이가장효율적인방법일것으로평가받고있다 (Chung et al. 2009). 특히국내한약재시장의대부분을차지하는인삼 (Panax ginseng) 에서는 DNA 분석법을위한분자마커개발연구가활발하게진행되고있으며, 종류로는 random amplified polymorphic DNA(RAPD) (Boehm et al. 1999; Kim and Choi 2003), amplified fragment length polymorphism (AFLP) (Kim et al. 2005), restriction fragment length polymorphism (RFLP) (Um et al. 2001), internal transcribed spacer/non-transcribed spacer (ITS/NTS) (Cui et al. 2003; Ngan et al. 1999), simple sequence repeat (SSR) (Bang et al. 2011a, b; Kim et al. 2007; Nguyen et al. 2010) 그리고 sequence This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
152 J Plant Biotechnol (2016) 43:151 156 characterized amplified region (SCAR) (Choi et al. 2007; Choo et al. 2009; Lee et al. 2011) 를들수있다. 또한최근에는 next generation sequencing (NGS) 유전체분석기반기술을이용한유연관계분석결과가보고되었다 (Kim et al. 2015). 그렇기때문에인삼의경우와마찬가지로유전체분석기반을통한약용작물의유전육종시스템도입은기원정립뿐만아니라전통육종과상호조화를이루어계통선발및신품종육성의가속화와체계화가이루어질수있을것으로전망된다. 본리뷰논문에서는 Angelica 속식물의기원정립을위한형태학적연구, 세포분류학적연구및분자생물학적연구에대한국내외연구현황특히, 유전육종기반에관한최근의연구결과에대해정리하고어떻게활용될수있는지서술하고자한다. Angelica 속식물의국내외연구동향 Angelica 속은약 120 여종이포함된비교적큰속으로북반구지역에널리분포한다 (Choi et al. 2005). 특히한국, 중국, 일본을포함한동북아시아지역이 Angelica 속식물들의자생지이며종다양성이높게나타나는것으로알려져있다 (Vasil eva and Pimenov 1991). 한국에서자생하거나한약재로유통되는 Angelica 속식물은 18 종으로 Table 1 에나타내었다. 특히참당귀, 중국당귀, 일당귀가혼용되기도하는데한국에서자생하는것은참당귀와일당귀뿐이며중국당귀는우리나라기후환경이맞지않아재배 가되지않음이보고된바있다 (Yu et al. 2004). 하지만식의약소재로사용되는참당귀, 중국당귀, 일당귀 3 종은형태학적특성이분명히다름에도불구하고한약재에서혼용하여사용되는경우가많기때문에이들을분류하기위한형태학적연구, 세포분류학적연구및분자생물학적연구등이계속적으로보고되어왔다. 형태학적연구 형태학적연구에는참당귀, 중국당귀, 일당귀의잎과엽병및뿌리에대해해부형태적특징을관찰하여당귀기원및국내유통당귀류약재의판별에활용하고자하였다 (Sung et al. 2004). 그결과당귀류약재판별에있어서해부형태적특징을정리하여간단한동정 Key 를적용하여만든검색표가유용하게활용될수있음을보고하였다. 그리고 A. archangelica, A. palustris, A. sylvestris 3 종의뿌리, 줄기, 잎조직에 hand microtome 을이용하여해부학적특징을관찰한결과가보고되었다 (Lobiuc et al. 2012). 또한국내연구자들에의해근적외선분광법, 전자코및엑스선형광법과같은접근방식을통해참당귀, 중국당귀, 일당귀를분류한연구도보고되었다 (Cho et al. 2002). 하지만이들형태학적연구는식물체가성장한이후관찰할수있으며식물체가온전히유지되어야관찰하여분류할수있는기법으로써한약재로가공하여유통되는당귀류에대해서는분류가어렵다는결론을얻을수있다. Table 1 List of Angelica species in Korea Species Angelica sinensis Angelica acutiloba (Siebold & Zucc.) Kitag. Angelica anomala Ave-Lall. Angelica cartilaginomarginata Angelica czernaevia (Fisch. & C.A.Mey.) Kitag. Angelica dahurica (Fisch) Benth. et Hooker f. Angelica decursiva (Miq.) Franch. & Sav. Angelica gigas Nakai Angelica genuflexa Nutt. ex Torr. & A.Gray Angelica polymorpha Maxim. Angelica purpuraefolia Chung Angelica pubescens F.biserrata Angelica japonica A.Gray Angelica tenuissima Nakai Angelica florenti Franch. & Sav. ex Maxim. Angelica jaluana Nakai Korean name 중국당귀일당귀개구릿대처녀바디잔잎바디구릿대바디나물참당귀일천궁궁궁이지리강활중치모당귀갯강활고본두메당근삼수구릿대
J Plant Biotechnol (2016) 43:151 156 153 세포분류학적연구 Angelica 속의세포분류학적연구는 Angelica 속식물의기본염색체수가 2n=22 로알려지면서시작되었다 (Darlington and Wylie 1955). 이후한국에분포하는 Angelica 속식물인참당귀, 바디나물및일당귀에대한세포유전학적연구가보고되었고 (Koo et al. 2003; To, 1970) 참당귀, 중국당귀, 일당귀, 고본 (A. tenuissima), 바디나물 (A. decursiva), 구릿대 (A. dahurica) 와일천궁 (A. genuflexa) 에대한세포유전학적연구가수행되었다 (Choi et al. 2005). 일본에서는일당귀를비롯한구릿대, 바디나물, 참당귀, 일천궁등에서염색체수와핵형에대한연구가수행된바있으며 (Arano and Saito 1979; Hatano et al. 1975) 중국과러시아에서식하는 Angelica 속식물들에서도핵형에대한보고 (Pan et al. 1985; Vasil eva and Pimenov 1991) 가있다. 선행연구자들의보고에따르면 Angelica 속식물들의넓은분포지와지리적차이로인하여일부종에서세포유전적차이가관찰된다고하였다. 이처럼세포유전학적분류방법으로도모든 Angelica 속식물들을커버할수없으며같은종이라고하여도자생지의환경조건에따라변이가생길수있기때문에기초데이터로써의활용가치는낮다고보여진다. 분자생물학적연구 유전자분석이산지나생육환경에영향을받지않아한약재종감별에자주이용되고있다. 당귀속식물의분자적연구는 Mizukami 등 (1997) 이일당귀에서 5S-rDNA ITS 영역을분석한것에서시작되었다. Chen 등 (2010) 은 753 속 4,800 종에해당하는시료에대하여 ITS2 영역의염기서열을분석한결과 92.7% 에해당하는종의구분이가능하여약용식물의종간구분을위한범용바코드로지정하는것을제시한바있다. 또한 Yao et al. (2010) 도 50,790 종의식물과 12,221 종의동물에대하여 ITS2 영역의염기서열을비교분석한결과 67 ~ 91% 까지종의구분이가능하였으며 ITS2 영역의 secondary structure 를이용하면범용으로사용될수있는식물용 barcode 로의사용가능성을제시하였다. 이후연구자들은참당귀, 중국당귀, 일당귀의동정을위해 5S-rRNA spacer domains 의염기서열을제시한바있다 (Lee and Rasmussen 2000; Zhao et al. 2003). He 등 (2011) 도 Angelica 속에속하는다양한종에대하여 ITS 지역의염기서열을비교하여계통수를제작하여유연관계를분석하였다. 특히이연구를통하여중국의시중에서말린뿌리상태로유통되어구분이어려운당귀시료들을시중에서수집하여조사한결과일부위품의판별이가능하였다. Kim 등 (2012) 도 ITS 지역을이용하여분석한결 과참당귀와일당귀가중국당귀보다유연관계가가까운것으로조사되었으며, 한 중 일 3 국에서각기다른기원종으로규정하고있는당귀류의구별을위해시중에서유통중인당귀를종별로구입하여미각패턴과 ITS DNA 의염기서열을비교한결과일치하는결과를얻었다고보고하였다. 또한연구자들은중국당귀와혼합된재료들사이에서중국당귀를선발하는실험을수행하였고 ITS 분석으로이를효과적으로분리해낼수있다고하였다 (Feng et al. 2010). Choi 등은 ITS 부위의염기서열과 AFLP 분석으로 Angelica 속한약재감별을시도하였으며 (Choi et al. 2004), RAPD 를이용하여참당귀내에서내추대성과추대성품종을구분할수있었으며건재약재로사용되는참당귀와중국당귀그리고일당귀를판별할수있는 primer 를선발할수있었다 (Bang et al. 2002). 참당귀, 중국당귀, 일당귀를구별할수있는내부형태적특성과 5 개의 RAPD 프라이머를선발하였으며유연관계를분석한결과, 중국당귀와일당귀가근연관계에있음을보고하였다 (Lee et al. 2000). Mei 등 (2015) 도지역수집종인중국당귀와일당귀에대하여 RAPD 와 ISSR 을이용하여분석한결과를보고하였다. 또한중치모당귀 (A. pubescens) 와독활 (Aralia continentalis) 을분류하기위하여내부형태적특성과 RAPD 분석을병행하여연구한보고 (Lee et al. 2001) 가있다. 이처럼최근에는형태학적, 세포학적및분자생물학적판별기법을병행하여기원을판별하는연구가다수보고되고있다. Liao 등 (2013) 은핵 rdna 의 ITS 영역, EST 염기서열및 cpdna (rps16 intron, rps16-trnk, rpl32-trnl, trnl-trnt) 와종자구조형태적특성조사결과를이용하여 4 개의 Angelica 속식물 -A. hirsutiflora, A. oncosepala, A. paeoniifolia, A. sinensis- 에대하여분류한결과를보고하였다. 그리고 Gao 등 (2001) 은지역간차이가있는중국당귀 18 개를수집하여 RAPD 분석을통해변이를보이는밴드패턴을확인한바있다. 최근 Lu 등 (2015) 은 NGS 기법을통하여중국당귀에서 SSR 마커를개발하여 8 개종에대해분석하였다. 중국내에서중국당귀는 Female ginseng 이라고하여중국내한약재들사이에인삼과같이중요한자원으로가치를인정받고있으며분자생물학적기초연구가활발하게이루어지고있다. 하지만참당귀는국내에서중요한한약재로사용함에도불구하고연구가미흡한실정이다. 따라서참당귀의전체유전체에골고루분포하며, 이를분류동정하기위한다양한분자마커개발이계속적으로수행되어야한다. 결론 현재우리나라는약용작물생산의안정화, 재배기술의표준화, 품종의다양화, 한약재원료의혼 오용방지등을
154 J Plant Biotechnol (2016) 43:151 156 위해서과학적이며체계적인육종프로그램의도입이절실하다. 예를들어비교적세대진전과교배집단작성이용이한채소작물들의경우, 이미유전체기반의육종프로그램이활성화되는시점에있다. 이는최근염기서열해독장비및기술의발달로고품질의표준유전체서열이대량으로생산됨으로써염기서열기반의다양한분자마커들이발굴되고있기때문이기도하다. 따라서채소작물에서는이들을활용한다양한작물에서고밀도유전자연관지도가일부작성되었거나작성될것이며, 또한빠른시일내에분자마커기반의유전자연관지도와염기서열기반의물리지도가통합되어다양하게활용할수있을것이다. 이처럼다양한작물의통합유전자지도를활용한다면식물학적생육특성및품질관련유전자들에대한연관분석등이비교적용이하게이루어질것이다. 더욱이표준유전체서열을기반으로다양한품종및교배집단들에대한유전체해독, 비교유전체분석, genotyping by sequencing(gbs) 등을통해품종및계통특이적마커발굴및형질연관마커발굴, 유전자기능연구등이이루어질것으로전망된다. 선행연구자들은이미표준유전체기반의마커를개발하여녹두 (mungbean), 파 (bunching onion) 육종에활용하고있다 (Gupta et al. 2014; Tsukazaki et al. 2006; Yang et al. 2015). 현재국내에서는다부처유전체사업을토대로다양한작목에서유전체, 전사체및대사체분석이이루어지고있으며, 이로인해유전자기능및유전자연관지도작성의기초기반을마련할수있을것이다. 유전체및전사체분석을통해확보한대규모 single nucleotide polymorphism (SNP), insertion and deletion (InDel) 및 SSR 등의다형성분자마커를이용한고밀도연관및물리지도작성이이루어지고있고, 궁극적으로통합지도작성이이루어지게될것이다. 따라서가까운미래에참당귀특이주요농업형질연관유전자의정확도높고효율적인분자표지선발육종 (marker-assisted selection breeding system, MAS) 이이루어질것이다. 결론적으로참당귀와중국당귀에대한식별및분류체계가확립되어야하고당귀이외의약용작물도수입량에의존적인품목이있기때문에기원정립과판별에대한활발한연구가계속적으로진행되어야할것이다. 적요 약용식물자원은천연화장품소재, 제약산업그리고인간의건강을위한기능성식품의개발로확대된이후중요한자산이되고있다. 그러나세계적으로약용식물의명칭을각각다르게표기하고있으며계통발생학적기원이명확하지않다는단점을지니고있다. 이때문에소 비자들은매우큰혼란을겪고있으며특히형태학적으로유사한식물의말린뿌리로유통될때에는전문가들도그들의기원을구별하기가매우어렵다. 이러한이유때문에이처럼광범위하고다양한작물의기원을식별하기위해분자표지기법을적용하여활용되고있다. 이리뷰에서본연구자들은 Angelica 종의분화에관한적합한 기원정립 을위한현재의연구성과들을정리했다. 결론적으로 Angelica 종의식별을위해개발된분자적연구에대해설명하고약용작물의유전체정보를활용하여그들의기원정립및판별에대해논의할것이다. 사사 본논문은농촌진흥청연구사업 (PJ01102201) 의지원에의해수행되었습니다. References Arano H, Saito H (1979) Cytological studies in family Umbelliferae. Ⅳ. Karyotypes in genus Angelica 2. La Kromosomo Ⅱ-15-16:417-426 Chung JW, Lee GA, Lee SS, Bang KH, Park CB, Park YJ (2009) Cultivar discrimination of Korean and Chinese boxthorn (Lycium chinense Mill. and Lycium barbarum L.) using SSR markers. Korean J Medicinal Crop Sci 17:445-451 Bang KH, Chung JW, Kim YC, Lee JW, Jo IH, Seo AY, Kim OT, Hyun DY, Kim DH, Cha SW (2011a) Development of SSR markers for identification of Korean ginseng (Panax ginseng C. A. Meyer.) cultivars. Korean J Medicinal Crop Sci 19: 185-190 Bang KH, Jo IH, Chung JW, Kim YC, Lee JW, Seo AY, Park JH, Kim OT, Hyun DY, Kim DH and Cha SW (2011b) Analysis of genetic polymorphism of Korean ginseng cultivars and foreign accessions using SSR markers. Korean J Medicinal Crop Sci 19:347-353 Bang KH, Yu HS, Koo DH, Cho JH, Park HW, Seong NS, Park SI, Kim HS (2002) Selection of RAPD marker to discriminate the bolting-resistant varieties and commercial dried medicinal materials of Angelica species. Korean J Medicinal Crop Sci 10:46-50 Boehm CL, Harrison HC, Jung G, Nienhuis J (1999) Organization of American and Asian ginseng germplasm using randomly amplified polymorphic DNA (RAPD) markers. J Am Soc Hortic Sci 124:252-256 Chen S, Yao H, Han J, Liu C, Song J, Shi L, Zhu Y, Ma X, Gao T, Pang X, Luo K, Li Y, Li X, Jia X, Lin Y, Leon C (2010) Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS One 5:1-8, e8613 Cho CH, Kim SJ, Kim HJ (2002) Comparative studies on the discrimination of Angelicae Gigantis Radix by near-infrared
J Plant Biotechnol (2016) 43:151 156 155 spectroscopy, electronic nose and X-ray fluorescence spectrometry. Yakhak Hoeji 46:161-167 Choi HW, Koo DH, Lee WK, Kim SY, Sung JS, Seong NS, Suh YB and Bang JW (2005) Cytogenetic analysis of seven Angelica species. Korean J Medicinal Crop Sci 13:118-121 Choi HW, Song H, Koo DH, Bang JW, Hur Y (2007) Molecular and cytological characterization of species-specific repetitive DNA sequences for Angelica acutiloba. Korean J Genetic 29: 503-511 Choi HY, Choi YJ, Lee JH, Ham Ih (2004) Sequencing analysis on the ITS region and AFLP analysis to identify dried medicinal Angelica species. Kor J Herbology 19:91-9 Choo BK, Moon BC, Ji Y, Kim BB, Choi G, Yoon T, Kim HK (2009) Development of SCAR markers for the discrimination of three species of medicinal plants, Angelica decursiva (Peucedanum decursivum), Peucedanum praeruptorum and Anthricus sylvestris, based on the internal transcribed spacer (ITS) sequence and random amplified polymorphic DNA (RAPD). Biol Pharm Bull 32:24-30 Cui XM, Lo CK, Yip Kl, Dong TTX, Tsim KWK (2003) Authentification of Panax notoginseng by 5S-rRNA spacer domain and random amplified polymorphic DNA(RAPD) analysis. Planta Med 69:584-586 Darlington CD, Wylie AP (1955) Chromesome atlas of flowering plants. The McMillan Co. New York, USA. pp.210 Feng T, Liu S, He XJ (2010) Molecular authentication of the traditional Chinese medicinal plant Angelica sinensis based on internal transcribed spacer of nrdna. Electron J Biotechn 13:9-10 Gao WY, Qin EQ, Xiao XH, Yu HM, Gao GY, Chen SB, Zhao Y, Yang S (2001) Analysis on genuineness of Angelica sinensis by RAPD. Chinese Tradition Herbal Drugs 10:926-929 Gupta SK, Bansal R, Gopalakrishna T (2014) Development and characterization of genic SSR markers for mungbean (Vigan radiate (L.) Wilczek). Euphytica 195:245-258 Hatano K, Nishioka I, Iwasa S (1975) Cytogenetical suties of Umbelliferous plants. Ⅲ. The karyotype analysis of Angelica species in Japan. Jpn J Pharmacol 29:10-21 He Y, Hou P, Fan G, Song Z, Liu H, Li Y, Zhang Y (2011) Internal transcribed spacers (ITS) identification of Angelica anomala Lallem Chuanbaizhi (in Chinese) cultivars collected in Sichuan and their molecular phylogenetic analysis with other Angelica L. species. J Med Plants Res 5:3653-3659 Kim BB, Jeong JH, Jung SJ, Yun DW, Yoon ES, Choi YE (2005) Authentification of Korean Panax ginseng from Chinese Panax ginseng and Panax quinquefolius by AFLP analysis. J Plant Biotechnol 7:81-86 Kim CK, Choi HK (2003) Genetic diversity and relationship in Korean ginseng (P. schinseng) based on RAPD analysis. Korean J Genetic 25:181-188 Kim J, Jo BH, Lee KL, Yoon ES, Ryu GH, Chung KW (2007) Identification of new microsatellite markers in Panax ginseng. Mol Cells 24:60-68 Kim K, Lee SC, Lee J, Lee HO, Joh HJ, Kim NH, Park HS, Yang TJ (2015) Comprehensive survey of genetic diversity in chloroplast genomes and 45S nrdnas within Panax ginseng species. PloS ONE 10:e0117159 Kim YH, Choi G, Lee HW, Lee GH, Chae SW, Kim YH, Lee MY (2012) Comparison of Angelica species roots using taste sensor and DNA sequencing analysis. Kor J Herbology 27:37-42 Koo DH, Kim SY, Bang KH, Seong NS, Bang JW (2003) Cytogenetic analysis of Angelica plants using feulgen staining and multicolor fluorescence in situ hybridization. Kor J Plant Biotechnol 30:123-127 Lee MY, Im SH, Ju YS, Han KS, Jeong GJ, An DG, Kang HC, Ko BS (2000) Discrimination of the three Angelica species using the RAPDs and internal root structure. Korean J Medicinal Crop Sci Sci 8(3):243-249 Lee MY, Ju YS, Kim HJ, Ko BS (2001) Discrimination of Aralia continentalis root by the random amplified polymorphic DNA analysis and morphological characteristics. Kor J Orient Med 7:145-152 Lee SB, Rasmussen SK (2000) Molecular markers in some medicinal plants of the Apiaceae family. Euphytica 114:87-91 Lee JW, Kim YC, Jo IH, Seo A, Lee JH, Kim OT, Hyun DY, Cha SW, Bang KH, Cho JH (2011) Development of an ISSRderived SCAR marker in Korean ginseng cultivars (Panax ginseng CA Meyer). J Ginseng Res 35:52-59 Liao C, Downie SR, Li Q, Yu Y, He X, Zhou B (2013) New insights into the phylogeny of Angelica and its allies (Apiaceae) with emphasis on east asian species, inferred from nrdna, cpdna, and Morphological Evidence. Syst Bot 38:266-281 Lobiuc A, Zamfirache MM, Ivănescu L (2012) Comparative anatomical investigations on some species of the genus Angelica L. Contributii Botanice 47 Lu Y, Cheng T, Zhu T, Jiang D, Zhou S, Jin L, Yuan Q, Huang L (2015) Isolation and characterization of 18 polymorphic microsatellite markers for the Female Ginseng Angelica sinensis (Apiaceae) and cross-species amplification. Biochem Syst Ecol 61:488-492 Mei Z, Zhang C, Khan A, Zhu Y, Tania M, Luo P, Fu J (2015) Efficiency of improved RAPD and ISSR markers in assessing genetic diversity and relationships in Angelica sinensis (Oliv.) Diels varieties of China. Electron J Biotechn 18:96-102 Ministry of Agriculture, Food and Rural Affairs(MAFRA) (2015) Production performance of industrial crops. Ministry of Agriculture, Food and Rural Affairs. Sejong, Korea Mizukami H, Hao BS, Tanaka T (1997) Nucleotide sequence of 5S-rDNA intergenic spacer region on Angelica acutiloba. Nat Med 51:376-378 Ngan F, Shaw P, But P, Wang J (1999) Molecular authentication of Panax species. Phytochemistry. 50:787-791 Nguyen VD, Nirala R, Choi SR, Uhm TS, Yang TJ, Ahn IO, Lim YP (2010) Development and characterization of new microsatellite markers in Panax ginseng C. A. Meyer from BAC end sequences. Conserv Genet 11:1223-1225 Pan ZH, Chin HC, Wu ZJ, Yuan CQ (1985) The karyotype of Angelica dahurica and their taxonomical significance (Umbelliferae). Acta Phytotax Sin 23:185-187
156 J Plant Biotechnol (2016) 43:151 156 Sung JS, Bang KW, Park CH, Park CG, Yu HS, Park HW, Seong NS (2004) Discrimination of Angelica radix based on anatomical characters. Korean J Medicinal Crop Sci 12:67-72 To CA (1970) Cytotaxonomical study of genus Angelica. J Kor Res Inst Better Living 5:57-61 Tsukazaki H, Fukuoka H, Song YS, Yamashita K, Wako T, Kojima A (2006) Considerable heterogeneity in commercial F1 varieties of bunching onion (Allium wstulosum) and the proposal of a breeding scheme for conferring genetic traceability using SSR markers. Breeding Sci 56:321-326 Um JY, Chung HS, Kim MS, Na HJ, Kwon HJ, Kim JJ, Lee KM, Lee SJ, Lim JP, Do KR, Hwang WJ, Lyu YS, An NH, Kim HM (2001) Molecular authentication of Panax ginseng species by RAPD analysis and PCR-RFLP. Biol Pharm Bull 24:872-875 Vasil eva MG, Pimenov MG (1991) Karyotaxomomical analysis in the genus Angelica (Umbelliferae). Plant Syst Evol 177:117-138 Yang L, Wen C, Zh ao H, Kiu Q, Yang J, Liu L, Wang Y (2015) Development of polymorphic genic SSR Markers by transcriptome sequencing in the welsh onion (Allium fistulosum L.). Appl Sci 5:1050-1063 Yao H, Song J, Liu C, Luo K, Han J, Li Y, Pang X, Xu H, Zhu Y, Xiao P, Chen S (2010) Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS ONE 5:e13102 Yu HS, Park CH, Park CG, Kim YG, Park HW, Seong NS (2004) Growth characteristics and tield of the three species of genus Angelica. Korean J Medicinal Crop Sci 12:43-46 Zhao KJ, Dong TTX, Tu PF, Song ZH, Lo CK, Tsim KWK (2003) Molecular genetic and chemical assessment of radix Angelica (Danggui) in China. J Agr Food Chem 51:2576-25