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木香、川木香、土木香、青木香和红木香药材的ITS2条形码分子鉴定

Keywords: DNA条形码,物种鉴定,ITS2序列,菊科,马兜铃科,五味子科

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Abstract:

通过分析木香、川木香、土木香、青木香、红木香药材的ITS2(internaltranscribedspacer2)条形码序列,探讨木香类药材鉴定新方法。对60份样品提取基因组DNA,PCR扩增ITS2序列并进行双向测序,所得序列经CodonCodeAligner拼接后,采用MEGA5.0软件进行序列比对,计算种内和种间遗传距离(K2P),构建邻接树(neighbor-joingtree,NJTree)。结果表明,无论是菊科的木香、川木香、土木香,还是马兜铃科的青木香和五味子科的红木香,ITS2序列种内变异均小于种间变异,ITS2序列所有单倍型比对后长度为272bp,变异位点达到162个;遗传距离显示物种种内平均K2P远远小于种间平均K2P;NJ树结果显示木香、川木香、土木香、青木香和红木香药材可明显区分,且能鉴别川木香的2个基原物种。因此,ITS2序列可用于鉴定木香、川木香、土木香、青木香、红木香药材,为临床安全用药提供依据。

References

[1]  中国药典.一部[S]. 2000:154.
[2]  王绪颖,贾晓斌,陈彦.木香类药材的研究进展[J]. 中药材,2010,33(1):153.
[3]  孙守祥.木香药材历史沿革中的基原变迁与分化[J].中药材,2008,31(7): 1093.
[4]  王小英.木香对大鼠实验性急性胃粘膜损伤的影响[J].中医研究,2004,17(2): 21.
[5]  韩坚,林煌权,钟志勇,等.木香超临界提取物抗实验性胃溃疡的研究[J].中药材,2005,28(11): 1017.
[6]  侯鹏飞,陈文星,赵新慧,等.木香挥发性成分气质联用分析及其抑制血小板聚集作用的研究[J].中国实验方剂学杂志,2008,14(7): 26.
[7]  沈映君. 中药药理学[M]. 北京: 人民卫生出版社, 2000: 553.
[8]  Lee M G, Lee K T, Chi S G, et al. Costunolide induces apoptosis by ROS mediated mitochondrial permeability transition and cytochrome C release[J]. Biol Pharm Bull, 2001, 24(3): 303.
[9]  王永兵,许华,王强,等. RP-HPLC法测定川木香中木香烃内酯和去氢木香内酯的含量[J].西北药学,2000,15(6): 250.
[10]  孟青,郭晓玲,詹靖,等.木香CO2超临界萃取物中木香烃内酯、去氢木香内酯的鉴别与含量测定[J].广东药学院学报,2001,17(4): 284.
[11]  刘圆,彭镰心,刘超,等.木香、川木香、土木香的高效液相色谱指纹图谱鉴别[J].药物分析杂志,2006,26(11): 1574.
[12]  胡晓炜, 徐爱仁. 三种木香的气相色谱法鉴别与系统聚类分析[J]. 中国现代应用药学杂志, 2005, 22(3): 249.
[13]  Shum K C, Chen F, Li S L, et al. Authentication of Radix Aucklandiae and its substitutes by GC-MS and hierarchical clustering analysis [J]. J Sep Sci, 2007, 30: 3233.
[14]  Chen S M,Fan M Y, Tseng C C, et al. Pharmacokinetics and nephrotoxicity of aristolochic acid in rabbits[J]. Toxicon, 2007, 50:180.
[15]  Wen Y J, Su T, Tang J W, et al. Cytotoxicity of phenanthrenes extracted from Aristolochia contorta in human proximal tubular epithelial cell line[J]. Nephron Exp Nephrol, 2006, 103: e95.
[16]  Li D Z, Liu J Q, Chen Z D, et al. Plant DNA barcoding in China [J]. J Syst Evol, 2011, 49: 165.
[17]  Kress W J, Wurdack K J, Zimmer E A, et al. Use of DNA barcodes to identify flowering plants[J]. Proc Natl Acad Sci USA, 2005, 102: 836.
[18]  Song J Y,Yao H,Li Y,et al. Authentication of the family Polygonaceae in Chinese pharmacopoeia by DNA barcoding technique[J].J Ethnopharmacol, 2009,124( 3): 434.
[19]  辛天怡,姚辉,罗焜,等.羌活药材ITS/ITS2 条形码鉴定及其稳定性与准确性研究[J].药学学报,2012,47(8): 1098.
[20]  韩建萍,李美妮,罗焜,等.DNA条形码鉴定洋金花及其伪品[J]. 药学学报,2011, 46 (11): 1408.
[21]  陈士林,郭宝林,张贵君,等.中药鉴定学新技术新方法研究进展[J].中国中药杂志,2012,37(8): 1043.
[22]  陈士林. 中药DNA 条形码分子鉴定[M]. 北京:人民卫生出版社,2012:4.
[23]  陈士林,姚辉,韩建萍,等.中药材 DNA 条形码分子鉴定指导原则[J].中国中药杂志,2013,38(2):141.
[24]  Chen S L, Yao H, Han J P, et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS ONE, 2010, 5: e8613.
[25]  Yao H, Song J Y, Liu C, et al. Use of ITS2 region as the universal DNA barcode for plants and animals[J]. PLoS ONE, 2010, 5: e13102.
[26]  Keller A, Schleicher T, Schultz J, et al. 5.8S-28S rRNA interaction and HMM-based ITS2 annotation[J]. Gene, 2009, 430: 50.
[27]  中国药典.一部[S]. 2010.
[28]  Tamura K, Peterson D, Peterson N, et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011, 28: 2731.
[29]  Chen F, Chan H Y E, Wong K L, et al. Authentication of Saussurea lappa, an endangered medicinal material, by ITS DNA and 5S rRNA sequencing[J]. Planta Med, 2008,74: 889.
[30]  Ma X C, Xie C X, Guan M, et al. High levels of genetic diversity within one population of Rheum tanguticum on the Qinghai-Tibet Plateau have implications for germplasm conservatio[J]. Pharm Crops, 2014,5:1.
[31]  Song J Y, Shi L C, Li D Z, et al. Extensive pyrosequencing reveals frequent intra-genomic variations of internal transcribed spacer regions of nuclear ribosomal DNA[J]. PLoS ONE, 2012, 7(8): e43971.

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