基于叶绿体基因组SNP鉴定不同化学型的广藿香  

Identification of the Different Chemotypes of Pogostemon cablin Based on Chloroplast Genome SNP

作　　者：陆亚茹 彭晓祺 吴文如[1] 行冰楠 邹何元 LU Ya-ru;PENG Xiao-qi;WU Wen-ru;XING Bing-nan;ZOU He-yuan(School of Pharmaceutial Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《中药材》2024年第9期2192-2197,共6页Journal of Chinese Medicinal Materials

基　　金：中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”(2060302);广州中医药大学大学生创新创业训练计划项目(202210572232);广东省科学技术厅2020年度广东省科技特派员项目(KTP20200137)。

摘　　要：目的:基于叶绿体基因组单核苷酸多态性(SNP)位点的变异,建立一种区分不同化学型广藿香的分子鉴定方法。方法:根据不同化学型广藿香叶绿体基因组ycf3-trnS_GGA基因存在插入/缺失一个碱基的差异,设计扩增此段序列的引物,对不同广藿香样品进行PCR扩增和双脱氧测序分析。同步利用气相色谱法测定广藿香样品中广藿香酮和百秋李醇的含量并计算酮醇比值。结果:广藿香鲜品、药材及含广藿香的中成药均可扩增出196 bp条带,醇型广藿香在该序列的130 bp处为A碱基,酮型广藿香在该位点缺失A碱基,样品化学型的分子鉴定结果与气相色谱法验证的结果一致。结论:所建立的基于广藿香叶绿体基因组SNP位点的分子鉴定方法可有效区分不同化学型的广藿香,该方法准确可靠,具有推广应用的潜力。Objective:To establish a molecular identification method for different chemotypes of Pogostemon cablin based on the variation of single nucleotide polymorphism(SNP)in chloroplast genome.Methods:According to the difference of one base insertion/de-letion in ycf3-trnS_GGA gene of chloroplast genome of different chemotypes of Pogostemon cablin,primers were designed to amplify this sequence,and different samples of Pogostemon cablin were amplified by PCR and confirmed by dideoxy sequencing.The contents of pat-choulenone and patchouli alcohol in Pogostemon cablin were determined by gas chromatography and the ratio of them was calculated.Results:A band of 196 bp could be amplified from fresh,medicinal and proprietary Chinese medicines containing Pogostemon cablin.Alcohol type Pogostemon cablin was A 130 bp of the sequence,the alcohol type Pogostemon cablin was A base,while the ketone type lacked an A base at this site,and ketone type Pogostemon cablin was missing A base at this site.The molecular identification results of sample chemotypes was consistent with the results of gas chromatography.Conclusion:The established molecular identification method based on the SNP site of chloroplast genome of Pogostemon cablin can effectively distinguish different chemotypes of Pogostemon cablin,and this method is accurate,reliable,and has the potential to be widely applied.

关 键 词：广藿香 化学型 叶绿体基因组 单核苷酸多态性 气相色谱 

分 类 号：R282.71[医药卫生—中药学]