对市场上一种新的松贝伪品的比较研究

目的： 比较松贝与新出现伪品的不同特征，为鉴别检验提供依据。方法： 综合性状、聚合酶链式反应-限制性内切酶多样性（PCR-RFLP）法、DNA序列比对、系统进化树以及遗传距离等方法，寻找松贝与新伪品的鉴别要点。结果： 该贝母在性状上与松贝正品还是存在一定差异。按照《中华人民共和国药典》2015年版PCR-RFLP法检验，松贝PCR产物可以被完全酶切，而该伪品的PCR产物只能部分被酶切；测序分析可知，其部分酶切的原因是SmaⅠ酶切位点"CCCGGG"中第2位为C/T多态性位点。在内部转录间隔区（ITS区）266 bp范围内，该伪品与松贝比较有8个突变位点。系统进化树分析，该伪品与川贝母6个基原物种没有聚为一支。遗传距离分析，该伪品与川贝母6个基原物种的距离明显大于6个基原物种之间的距离。结论： 该贝母的基原植物与松贝不同，应与松贝严格区分，以维护消费者经济利益，并为安全用药提供保障。

Objective: To carry out comparative study on Songbei and its new adulterant,and to provide evidence for identification.Methods: Multiple analytical methods[appearance characteristics,polymerase chain reaction-restriction fragment length polymorphisms(PCR-RFLP),DNA sequence alignment,phylogenic analysis and heritage distance analysis]were utilized to identify Songbei and its new adulterant.Results: Some subtle difference in appearance characteristics could be found by carefully observation.The PCR product of the adulterant could be partially cutted according to the PCR-RFLP method in Pharmacopoeia of the People's Republic of China(Chinese Pharmacopoeia)2015,while the PCR product of Songbei could be completely cutted.By sequencing analysis, a "C/T" polymorphism was found on the second position of SmaⅠ restriction site "CCCGGG",which might be the reason why the PCR product was only partially cutted.There were 8 mutation sites within the length of 266 bp on the internal transcribed spacer(ITS)region of the adulterant,compared to that of Songbei.Phylogenic analysis of neighbor-joining(N-J)tree showed that the adulterant failed to be clustered as one branch with the 6 kinds of originating plants of Fritillariae Cirrhosae Bulbus.The heritage distance analysis revealed a much greater distance between the adulterant and the 6 kinds of originating plants of Fritillariae Cirrhosae Bulbus,compared to the distance within the 6 kinds of originating plants.Conclusion: The origin of the adulterant is verified to be different from that of Songbei and should be strictly identified to assure the interest of consumers and the safety medicinal treatment.

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