RAPD和ISSR分子标记检测大麻的遗传多样性初探

目的利用随机扩增多态性DNA和简单序列重复区间扩增分子标记检测大麻遗传多样性,并探讨其在法医学中的应用价值。方法收集中国4省6个地区的100株大麻叶子样品,采用CTAB法提取基因组DNA,设计选择11个RAPD引物和13个ISSR引物,采用6%中性聚丙烯酰胺凝胶电泳-硝酸银染色法进行检测,根据出现的条带数目和片段大小等分析大麻的多样性。结果 11条RAPD引物扩增出的片段在200bp以上共52条,其中具有多态性的27条;ISSR引物扩增出126条,其中具有多态性的73条;多态性条带比率分别为51.9%和57.9%,其差异不具有统计学意义(P>0.05)。结论 RAPD和ISSR两种方法均可用于大麻遗传多样性分析,对检测毒品原植物的种类和来源地具有一定的应用前景。

Investigation of genetic diversity of hemp cultivars using RAPD and ISSR molecular markers

Objective To investigate the genetic diversity of hemp cultivars from six districts of four provinces in China by using randomly amplified polymorphic DNA(RAPD) and inter-simple sequence repeat(ISSR) molecular markers,and evaluate its application value.Methods The genomic DNA of 100 hemp cultivars was extracted by hexadecyltrimethy ammonium bromide method(CTAB).We designed 23 RAPD and 30 ISSR primers and the amplified products were detected by 6% polyacrylamide gel electrophoresis followed by silver-stain method.Genetic diversity of hemp cultivars was analyzed according to the number of bands and fragments size.Results 11 RAPD primers generated 52 bands which were over 200bp,of which 27 were polymorphic fragments.Also,13 ISSR primers generated 126 bands,of which 73 were polymorphic fragments.The polymorphism rate was 51.9% and 57.9% respectively,and there was no statistically significant(P>0.05).Conclusion Both RAPD and ISSR methods can be used for analysis of genetic diversity of hemp,which will have certain application prospect in identifying types and sources of drug plants.