基于ITS2序列的人参及同属易混品西洋参种子的分子鉴定
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摘要
目的基于DNA条形码技术建立人参与西洋参种子的分子鉴别方法。方法采用生药鉴定方法研究其性状、显微特征,并结合DNA条形码技术,基于中药材DNA条形码数据库,通过ITS2序列比对、遗传距离比较和构建临接(NJ)系统发育树,对人参与西洋参的种子进行鉴别研究。结果人参与西洋参种子的种内遗传距离分别小于种间遗传距离,系统发育树图中各物种均聚为一支,来自9个产地的42份人参及西洋参的种子均为正品,且容易区分。结论基于ITS2序列DNA条形码技术可以快速、准确、有效地鉴别人参与西洋参种质资源。
Objective Traditional identification methods of pharmacognosy is difficult to distinguish the seeds of Panax ginseng and Panax quinquefolius. In order to improve the efficacy and accuracy of identification and provide the scientific foundation for the establishment of Chinese herbal medicine seed quality standards, molecular identification methods of the seeds were established by DNA barcoding technology. Methods The pharmacognostical identification method was used to study the morphological identification and microscopic characters of different seeds. DNA barcodes and Chinese Pharmacopoeia species standard barcode database were employed to identify the seeds by ITS2 sequence comparison, genetic distance comparison and systematic NJ tree construction. Results Intraspecific genetic distances of individuals participating were smaller than interspecific genetic distances. Phylogenetic tree map showed that two species were repectively clustered into one. A total of 42 samples of seeds from Panax ginseng and Panax quinquefolius produced by nine areas were all top-quality and easy to distinguish. Conclusion ITS2 DNA barcodes can identify and differentiate the seeds of P. ginseng and P. quinquefolius germplasm resources quickly, accurately and efficiently.
Objective Traditional identification methods of pharmacognosy is difficult to distinguish the seeds of Panax ginseng and Panax quinquefolius. In order to improve the efficacy and accuracy of identification and provide the scientific foundation for the establishment of Chinese herbal medicine seed quality standards, molecular identification methods of the seeds were established by DNA barcoding technology. Methods The pharmacognostical identification method was used to study the morphological identification and microscopic characters of different seeds. DNA barcodes and Chinese Pharmacopoeia species standard barcode database were employed to identify the seeds by ITS2 sequence comparison, genetic distance comparison and systematic NJ tree construction. Results Intraspecific genetic distances of individuals participating were smaller than interspecific genetic distances. Phylogenetic tree map showed that two species were repectively clustered into one. A total of 42 samples of seeds from Panax ginseng and Panax quinquefolius produced by nine areas were all top-quality and easy to distinguish. Conclusion ITS2 DNA barcodes can identify and differentiate the seeds of P. ginseng and P. quinquefolius germplasm resources quickly, accurately and efficiently.
引文
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[12]孙涛,孔德英,滕少娜,等.基于ITS2条形码的人参属物种鉴定研究[J].湖北农业科学,2016(18):5072-5074.
[13]张宝华.聚焦植物石蜡切片制作[J].吉林省教育学院学报,2013,29(4):153-154.
[14]Mao R,Xia P,He Z,et al.Identification of seeds based on molecular markers and secondary metabolites in Senna obtusifolia and Senna occidentalis[J].Bot Stud,2017,58(1):43-49.
[15]崔占虎,李越,袁庆军,等.黄芪与其混伪品的ITS序列分子鉴定研究[J].中国中药杂志,2012,37(24):1314-1319.
[16]代晓蕾.西洋参种子发育的形态及生理生化研究[D].南京:南京农业大学,2012.
[2]方海兰,夏从龙,段宝忠,等.基于DNA条形码的中药材种子种苗鉴定研究--以重楼为例[J].中药材,2016,39(5):986-990.
[3]李隆云,彭锐,李红莉,等.中药材种子种苗的发展策略[J].重庆中草药研究,2011,35(1):247-252.
[4]王鹏.西洋参与其混淆品人参的比较鉴别[J].中国药业,2015,23(6):90-91.
[5]Lo Y T,Li M,Shaw P C.Identification of constituent herbs in ginseng decoctions by DNA markers[J].Chin Med,2015,10(1):1-8.
[6]凃媛,熊超,师玉华,等.细小种子类毒性药材天仙子的DNA条形码鉴定[J].世界科学技术-中医药现代化,2014,16(11):2337-2342.
[7]罗红婷.人参、大黄等六种中药材显微量化研究[D].广州:广州中医药大学,2014.
[8]向婷婷,周志勇,刘朝奇,等.DNA条形码鉴定药用植物的研究进展[J].时珍国医国药,2015,26(1):181-183.
[9]Yang J,Dong L L,Wei G F,et al.Identification and quality analysis of Panax notoginseng and Panax vietnamensis var.fuscidicus through integrated DNAbarcoding and HPLC[J].Chin Herb Med,2018,10(2):176-182.
[10]陈士林,庞晓慧,姚辉,等.中药DNA条形码鉴定体系及研究方向[J].世界科学技术-中医药现代化,2011,13(5):747-754.
[11]宋晓娜,顾选,刘春生,等.基于trnL-trnF序列的人参和西洋参DNA条形码鉴定研究[J].中国中药杂志,2015,40(10):1914-1918.
[12]孙涛,孔德英,滕少娜,等.基于ITS2条形码的人参属物种鉴定研究[J].湖北农业科学,2016(18):5072-5074.
[13]张宝华.聚焦植物石蜡切片制作[J].吉林省教育学院学报,2013,29(4):153-154.
[14]Mao R,Xia P,He Z,et al.Identification of seeds based on molecular markers and secondary metabolites in Senna obtusifolia and Senna occidentalis[J].Bot Stud,2017,58(1):43-49.
[15]崔占虎,李越,袁庆军,等.黄芪与其混伪品的ITS序列分子鉴定研究[J].中国中药杂志,2012,37(24):1314-1319.
[16]代晓蕾.西洋参种子发育的形态及生理生化研究[D].南京:南京农业大学,2012.
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