DNA条形码技术在毒品原植物大麻鉴定中的应用

Application of DNA barcoding in cannabic identification

关键词:  DNA条形码  大麻  种属鉴定

Key words:  DNA barcoding  cannabis  species identification

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.