药学学报, 2013, 48(12): 1850-1855
引用本文:
孙稚颖, 庞晓慧. 基于DNA条形码技术探讨板蓝根及大青叶基原物种问题[J]. 药学学报, 2013, 48(12): 1850-1855.
SUN Zhi-ying, PANG Xiao-hui. Discussion on the botanical origin of Isatidis Radix and Isatidis Folium based on DNA barcoding[J]. Acta Pharmaceutica Sinica, 2013, 48(12): 1850-1855.

基于DNA条形码技术探讨板蓝根及大青叶基原物种问题
孙稚颖1, 庞晓慧2
1. 山东中医药大学药学院, 山东 济南 250355;
2. 中国医学科学院、北京协和医学院药用植物研究所, 濒危药材繁育国家工程实验室, 北京 100193
摘要:
本文对我国板蓝根及大青叶原植物的物种进行了探讨,以澄清分类上的混乱现象。利用PCR直接测序法对采集的全国22份样本,测定了核基因ITS2区和叶绿体matK基因片段,所得序列经CodonCode Aligner拼接后,用软件MEGA4.0进行相关数据分析,并构建NJ(邻接)树。 结果表明,所研究样本ITS2片段长度为191 bp,测序峰图显示不同研究样本间具有多个单核苷酸多态性(SNP)位点,部分样本具有杂合位点;从构建的NJ树图可以看出,基于核ITS2片段,所研究样本被分成两大支,其中一支与欧洲菘蓝相聚。基于叶绿体matK序列,所研究样本也被明显分成两支。因此本研究支持板蓝根及大青叶的原植物为菘蓝(Isatis indigotica Fortune),与欧洲菘蓝(Isatis tinctoria L.)是独立的两个物种,不支持Flora of China中将二者合并的观点。
关键词:    菘蓝      欧洲菘蓝      DNA条形码      分类     
Discussion on the botanical origin of Isatidis Radix and Isatidis Folium based on DNA barcoding
SUN Zhi-ying1, PANG Xiao-hui2
1. College of Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan 250355, China;
2. National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
Abstract:
This paper aimed to investigate the botanical origins of Isatidis Radix and Isatidis Folium, and clarify the confusion of its classification. The second internal transcribed spacer (ITS2) of ribosomal DNA, the chloroplast matK gene of 22 samples from some major production areas were amplified and sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. Phylogenetic study was performed using MEGA 4.0 software in accordance with the Kimura 2-Parameter (K2P) model, and the phylogenetic tree was constructed using the neighbor-joining methods. The results showed that the length of ITS2 sequence of the botanical origins of Isatidis Radix and Isatidis Folium was 191 bp. The sequence showed that some samples had several SNP sites, and some samples had heterozygosis sites. In the NJ tree, based on ITS2 sequence, the studied samples were separated into two groups, and one of them was gathered with Isatis tinctoria L. The studied samples also were divided into two groups obviously based on the chloroplast matK gene. In conclusion, our results support that the botanical origins of Isatidis Radix and Isatidis Folium are Isatis indigotica Fortune, and Isatis indigotica and Isatis tinctoria are two distinct species. This study doesn't support the opinion about the combination of these two species in Flora of China.
Key words:    Isatis indigotica    Isatis tinctoria    DNA barcodes    classification   
收稿日期: 2013-06-10
基金项目: 国家高技术研究发展计划(863计划)(2012AA021602);教育部长江学者和创新团队发展计划(IRT1150);中央级公益性科研院所基本科研业务费专项(YZ-12-08).
通讯作者: 庞晓慧
Email: xhpang@implad.ac.cn
相关功能
PDF(2341KB) Free
打印本文
0
作者相关文章
孙稚颖  在本刊中的所有文章
庞晓慧  在本刊中的所有文章

参考文献:
[1] Chinese Pharmacopoeia Commission. The Pharmacopoeia of the People's Republic of China. PartⅠ (中华人民共和国药典 第一部)[S]. Beijing: China Medical Science Press, 2010: 20, 191.
[2] Zhou TY, Lu LL, Yang G, et al. Brassicaceae[M]//In Wu ZY, Raven PH. Flora of China (Vol. 8). Beijing: Science Press; St. Louis: the Missouri Botanical Garden Press, 2001.
[3] Lou ZC, Qin B. Species Systematization and Quality Evaluation of Commonly Used Chinese Traditional Drugs (North-China Edition, Vol. 1) (常用中药材品种整理和质量研究 (北方编第1册) )[M]. Beijing: Beijing Medical University Press, 1995: 253-303.
[4] Xiao JH, Xiao H, Huang DW. DNA barcoding: new approach of biological taxonomy[J]. Acta Zool Sin (动物学报), 2004, 50: 852-855.
[5] Ren BQ, Chen ZD. DNA barcoding plant life[J]. Chin Bull Bot (植物学报), 2010, 45: 1-12.
[6] Hebert PDN, Ratnasingham S, de Waard JR. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species[J]. Proc Biol Sci, 2003, 270: S96-99.
[7] Schindel DE, Miller SE. DNA barcoding: a useful tool for taxonomists[J]. Nature, 2005, 435: 17.
[8] Chen SL, Song JY, Yao H, et al. Strategy and key technique of identification of Chinese herbal medicine using DNA barcoding[J]. Chin J Nat Med (中国天然药物), 2009, 7: 322-327.
[9] Chen SL, Yao H, Song JY, et al. Use of DNA barcoding to identify Chinese medicinal materials[J]. World Sci Technol Mod Tradit Chin Med (世界科学技术-中医药现代化), 2007, 9: 7-12.
[10] Ning SP, Yan HF, Gang H, et al. Current advances of DNA barcoding study in plants[J]. Biodiv Sci (生物多样性), 2008, 16: 417-425.
[11] Chen SL, Yao H, Han JP, et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS One, 2010, 5: e8613.
[12] Pang XH, Song JY, Zhu YJ, et al. Applying plant DNA barcodes for Rosaceae species identification[J]. Cladistics, 2011, 27: 165-170.
[13] Yao H, Song JY, Liu C, et al. Use of ITS2 region as the universal DNA barcode for plants and animals[J]. PLoS One, 2010, 5: e13102.
[14] Gao T, Yao H, Song JY, et al. Evaluating the feasibility of using candidate DNA barcodes in discriminating species of the large Asteraceae family[J]. BMC Evol Biol, 2010, 10: 324.
[15] Song JY, Shi LC, Li DZ, et al. Extensive pyrosequencing reveals frequent intra-genomic variations of internal transcribed spacer regions of nuclear ribosomal DNA[J]. PLoS One, 2012, 7: e43971.
[16] Chen SL. Molecular Identification of Traditional Chinese Medicine Based on DNA Barcoding (中药DNA条形码分子鉴定)[M]. Beijing: People's Medical Publishing House, 2012.
[17] Kress WJ, Wurdack KJ, Zimmer EA, et al. Use of DNA barcodes to identify flowering plants[J]. Proc Natl Acad Sci USA, 2005, 102: 8369-8374.
[18] Lahaye R, van der Bank M, Bogarin D, et al. DNA barcoding the floras of biodiversity hotspots[J]. Proc Natl Acad Sci USA, 2008, 105: 2923-2928.
[19] Sass C, Little DP, Stevenson DW, et al. DNA barcoding in the Cycadales: testing the potential of proposed barcoding markers for species identification of Cycads[J]. PLoS One, 2007, 2: e1154.
[20] Zhu YJ, Chen SL, Song JY, et al. DNA barcoding the medicinal plants of the genus Paris[J]. Acta Pharm Sin (药学学报), 2010, 45: 376-382.
[21] Keller A, Schleicher T, Schultz J, et al. 5.8S-28S rRNA interaction and HMM-based ITS2 annotation[J]. Gene, 2009, 430: 50-57.
[22] Wang XP, Zhou TY. Scientific research on Isatis indigotica[J]. Chin Tradit Herb Drugs (中草药), 1982, 13: 37-38.
[23] Qiao CZ, Cui X. Study on the identification of Isatis indigotica and Isatis tinctoria[J]. Acta Phytotaxon Sin (植物分类学报), 1984, 22: 237.
[24] Zhou Z, Ren DQ, Peng WQ. Preliminary report on the introduction and cultivation of Isatis indigotica and Isatis tinctoria[J]. J Chin Med Mater (中药材), 1994, 17: 6-8.
[25] Zhang LP, Wang B, Ren XS, et al. Investigation of germplasm resources of Radix Isatidis produced at Bozhou in Anhui province[J]. China J Chin Mater Med (中国中药杂志), 2004, 29: 1127-1130.
[26] Mohn T, Hamburger M. Glucosinolate pattern in Isatis tinctoria and I. indigotica seeds[J]. Planta Med, 2008, 74: 885-888.
[27] Spataro G, Taviani P, Negri V. Genetic variation and population structure in a Eurasian collection of Isatis tinctoria L.[J]. Gen Res Crop Evol, 2007, 54: 573-584.
[28] Gilbert (nee Stoker) G, Garton S, Karam A, et al. A high degree of genetic diversity is revealed in Isatis spp. (dyer's woad) by amplified fragment length polymorphism (AFLP)[J]. Theor Appl Genet, 2002, 104: 1150-1156.
[29] Cao H, Cai JN, Liu YP, et al. Correlative analysis between geographical distribution and nucleotide sequence of chloroplast matK gene of Cnidium monnieri fruit in China[J]. Acta Pharm Sin (药学学报), 2001, 36: 373-376.