鉴别烟草属植物的特异PCR引物研究

Study on specific PCR primers for identification of Nicotiana plants

【目的】为科学、高效鉴别检测涉案制烟原料中是否含有烟草材料。【方法】利用公共数据库中烟碱代谢途径相关基因序列和茄科基因组数据(含烟草未公开数据)开发分子标记,通过对3个烟草亚属11个组的91份烟草材料和17份非烟草材料进行实验验证,获得烟草属特异性扩增分子标记。【结果】从209对新开发的引物中筛选出2对稳定、可靠且具有烟草属特异性的标记,在91份烟草材料中均有目的扩增片段,而在17份非烟草材料中无扩增产物。【结论】上述2对烟草属特异性标记实现了烟草与非烟草材料的快速、有效鉴别检测,且方法简便易行、操作性强,可应用于烟草行业打假缉私工作中涉案制烟原料的鉴别检验。

To scientifically and efficiently identify and detect the tobacco materials contained in the tobacco raw materials involved in the case, molecular markers were developed using the sequence data of genes related to nicotine metabolism pathway and genomic data of Solanaceae(including unpublished tobacco genomic data) in public database in this study. The Nicotiana-specific markers were obtained by experimentally verifying 17 non-tobacco materials and 91 tobacco materials belonging to 11 sections of 3 sub-genus. The results of the study showed that 2 pairs of stable, reliable and Nicotiana-specific primers were screened out from 209 newly developed markers, which could clearly amplify the target length fragments in 91 Nicotiana materials, while there was no PCR product amplification in 17 non-Nicotiana materials. The method is simple and easy to operate, so it can be applied to the authenticity determination of tobacco materials in the anti-counterfeiting and anti-smuggling work of tobacco industry.