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DNA条形码技术在毒品原植物大麻鉴定中的应用(PDF)

《广西植物》[ISSN:1000-3142/CN:45-1134/Q]

期数:
2014年04期
页码:
552-556
栏目:
系统演化与生物多样性
出版日期:
2014-07-20

文章信息/Info

Title:
Application of DNA barcoding in cannabic identification
文章编号:
1000-3142(2014)04-0552-05
作者:
宋炳轲1 杨雪莹2 倪萍娅3 裴 黎2* 张 颖2 徐小玉2
1. 中国人民公安大学, 北京 100038; 2. 公安部物证鉴定中心, 北京 100038; 3. 杭州市公安局上城区分局, 杭州 311200
Author(s):
SONG Bing-Ke1 YANG Xue-Ying2 NI Ping-Ya3 PEI Li2* ZHANG Ying2 XU Xiao-Yu2
1. Chinese People’s Public Security University, Beijing 100038, China; 2. Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China; 3. ShangCheng District Branch of Hangzhou Public Security Bureau, Hangzhou 311200, China
关键词:
DNA条形码 大麻 种属鉴定
Keywords:
DNA barcoding cannabis species identification
分类号:
-
DOI:
10.3969/j.issn.1000-3142.2014.04.024
文献标识码:
A
摘要:
准确鉴定毒品原植物大麻的种属及品种具有重要的理论和实践意义。为了探讨DNA条形码技术用于毒品原植物大麻种属鉴定及品种鉴定的可行性,该研究以60份大麻原植物(分别采自内蒙、黑龙江、陕西延安、陕西榆林4个地区的栽培大麻雌雄各6株及新疆玛纳斯地区的野生大麻雌雄各6株)为材料,通过从其叶片中提取的DNA为模版,利用核糖体DNA基因间隔区的通用引物ITS2和叶绿体DNA的通用引物psbA-trnH进行PCR扩增,对扩增片段进行双向测序,将测序结果进行人工矫正和比对。结果显示:所有大麻样本的ITS2扩增片段序列没有变异完全一致,但psbA-trnH扩增片段变异较大共检测出8种cpDNA单倍型,用MEGE5.1软件计算种间遗传距离,并构建NJ系统聚类树可以有效把这五个地区的大麻样本区别开来,因此证明DNA条形码技术在毒品原植物大麻的种属鉴定方面具有可行性,但其用于大麻的种属鉴定的准确性、可靠性及在其来源地鉴定及品种鉴定中的可能性还有待进一步深入地研究。
Abstract:
To study the feasibility of DNA barcoding on cannabic species identification,DNA were extracted from sixty cannabis plants’ leaves(cultivated cannabis from Inner Mongolia,Heilongjiang,Shanxi Yan’an and Yulin,wild cannabis from Xinjiang Manas,six male and female cannabis from each region),used ribosomal DNA intergenic region ITS2 universal primers and cpDNA psbA-trnH primer for PCR amplification,then sequenced in both directions,the results compared to manual correction and finally for Blast comparison. ITS2’amplified sequences of all samples were completely consistent,but psbA-trnH’amplified sequences varied greatly,which were detected eight kinds of cpDNA haplotypes. MEGE 5.1 software was used to calculate the genetic distance between species,and build NJ phylogenetic trees which could effectively separate the five regions of cannabic samples. So DNA barcoding on cannabic species identification was proved to have viability,but the accuracy,reliability and possibilities of identification of its origin and species need further research.

参考文献/References

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Yang SJ(杨淑娟),Zhang YC(章英才),Zheng GQ(郑国琦). 2010. Comparative study on dissected structures of normal fruit and cracking ones of Lingwu Long-jujube(灵武长枣正常果与裂果解剖结构的比较研究)[J]. Northern Hortic(北方园艺),(22):15-18
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(灵武长枣多糖积累分布特征研究)[J]. Northern Hortic(北方园艺),(21):7-11Chen SL,Yao H,Han JP,et al. 2010. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS one,5(1):8 613
Kress WJ,Ericks DL. 2012. DNA Barcodes: Methods and Protocols[M]. Berlin:Springer,858:3-8
Li DZ,Liu JQ,Chen ZD,et al. 2011. Plant DNA barcoding in China[J]. J Syst & Evol,49(3):165-168
Li SJ(李仕金),Xin PY(辛培尧),Zhou J(周军),et al. 2008. Progresses on rsearch of sexual differentiation of Cannabis sativa L.(大麻性别研究进展)[J]. Plant Fiber Sci Chin(中国麻业学),30(2):110-113
Lu YX(卢延旭),Dong P(董鹏),Cui XG(崔晓光),et al. 2007. Difference between industrial hemp and marijuana hemp and industrial hempcs use value(工业大麻与毒品大麻的区别及其可利用价值)[J]. Chin Pharmacol Bull(中国药理学通报),23(8):1 112-1 114
Liu Y(刘莹),Zhang AS(张安世),Zhao LX(赵利新). 2013. The phylogenetic relationship of some bryophytes based on trnH-PsbA gene(基于trnH-PsbA序列的部分苔藓植物亲缘关系分析)[J]. Hubei Agric Sci(湖北农业科学),52(20):5 057-5 059
Naciri Y,Caetano S,Salamin N. 2012. Plant DNA barcodes and the influence of gene flow[J]. Mol Ecol Resour,12(4):575-580
Ren BQ(任保青),Chen ZD(陈之端). 2010. DNA barcoding plant life(植物DNA条形码技术)[J]. Chin Bull Bot(植物学报),45(1):1-12
Schoch CL,Seifert KA,Huhndorf S,et al. 2012. Nuclear ribosomal internal transcribed spacer(ITS)region as a universal DNA barcode marker for Fungi[J]. PNAS,109(16):6 241-6 246
Schaal BA,Hayworth DA,Olsen KM,et al. 1998. Phylogeographic studies in plants:problems and prospects[J]. Mol Ecol,7(4):465-474
Xie XM(谢幸媚),Wu KL(吴坤龙),Li JY(李俊亿),et al. 2000. The application and evaluation of plant DNA testing in criminal investigations(植物DNA鉴定在犯罪侦查上之应用与评估)[J]. Foren Sci(刑事科学),49:1-7
Yao H,Song J,Liu C,et al. 2010. Use of ITS2 Region as the universal DNA Barcode for plants and animals[J]. PLoSone,5:13 102
Zhu YJ(朱英杰),Chen SL(陈士林),Yao H(姚辉),et al. 2010. DNA barcoding the medicinal plants of the genus Paris(重楼属药用植物 DNA 条形码鉴定研究)[J]. Acta Pharm Sin(药学学报),45(3):376-382Maxim.(狗枣猕猴桃果实发育的解剖学观察)[J]. Chin Bull Bot(植物学通报),19(4):469-476
Chen YP(陈亚萍),Zhang YC(章英才),Jing HX(景红霞),et al. 2013. The characteristic of photosynthesis and the growth of fruit of Ziziphus jujuba Mill cv. lingwuchangzao(灵武长枣光合特性及果实生长发育特征)[J]. Northern Hortic(北方园艺),(19):1-4
Liu GC(刘国成),Ma HY(马怀宇),Lv DG(吕德国),et al. 2012. Research on the change of ‘Hanfu’apple fruit anatomic structure and quality during storage(“寒富”苹果贮藏期果实解剖结构及品质变化研究)[J]. Northern Hortic(北方园艺),(15):1-4
Wang XL(王勋陵). 1974. A preliminary study of the developmental anatomy of Chinese jujube fruit(枣果实发育解剖学的初步观察)[J]. Acta Bot Sin(植物学报),16(2):161-169
Xie MH(颉敏华),Zhang HM(张永茂),Li SQ(李守强),et al. 2008. Post harest fresh-keeping storage characteristics of‘lingwuchangzao(Ziziphus jujuba Mill. )(灵武长枣采后保鲜贮藏特性研究)[J]. Acta Bot Bor-Occ Sin(西北植物学报),28(5):1 031-1 035
Yang J(杨军),Zhang YC(章英才),Su WD(苏伟东),et al. 2011. Isolation and assaying of polysaccharide in Ziziphus jujuba Mill cv. lingwuchangzao(灵武长枣多糖含量的测定研究)[J]. Northern Hortic(北方园艺),(14):35-37
Yang SJ(杨淑娟),Zhang YC(章英才),Zheng GQ(郑国琦). 2010. Comparative study on dissected structures of normal fruit and cracking ones of Lingwu Long-jujube(灵武长枣正常果与裂果解剖结构的比较研究)[J]. Northern Hortic(北方园艺),(22):15-18
Yang SJ(杨淑娟),Zheng GQ(郑国琦),Zhang YC(章英才). 2011. Ultracytochemical localization of Ca2+ in the normal fruits and cracking ones of Lingwu Long-jujube(灵武长枣正常果及裂果中Ca2+的细胞化学定位研究)[J]. Acta Bot Bor-Occ Sin(西北植物学报),31(1):0084-0088
Yu JF(喻菊芳),Wei TJ(魏天军),Chen WJ(陈卫军). 2008. The research of germplasm resources and the study on variety selection of Ziziphus jujuba Mill cv. lingwuchangzao(灵武长枣种质资源调查和品种选优研究)[J]. Chin Fruits(中国果树),(1):56-57
Yu JF(喻菊芳),Zhu LC(朱连成),Wei TJ(魏天军). 2004. Ningxia(Lingwu)Long-jujube textual research(宁夏(灵武)长枣考证)[J]. Ningxia J Agric For Sci Technol(宁夏农林科技),(5):31-32
Zhang YC(章英才),Su WD(苏伟东),Yang J(杨军). 2012. The store localization characteristics of polysaccharide in Ziziphus jujuba Mill cv. lingwuchangzao(灵武长枣多糖积累分布特征研究)[J]. Northern Hortic(北方园艺),(21):7-11

备注/Memo

备注/Memo:
收稿日期: 2014-01-19
修回日期: 2014-03-08
基金项目: 公安部物证鉴定中心基本科研业务费专项(2010JB002)
作者简介: 宋炳轲(1987-),女,河南禹州市人,硕士研究生,主要从事法医遗传学的研究,(E-mail)sbk1987@126.com。 *通讯作者: 裴黎,主任法医师,硕士生导师,主要从事分子生物学与法医遗传学研究,(E-mail)peili128@sina.com。
更新日期/Last Update: 2014-07-20