摘要
目的:基于败酱类药材基原植物的叶绿体基因组序列分析开发特定的DNA条形码,准确鉴别败酱草及其近缘种.方法:采用Illumina高通量测序技术对败酱科败酱属植物白花败酱、黄花败酱和异叶败酱的叶绿体基因组进行测序;采用生物信息学软件对基因组进行组装注释、特征分析、序列比较和系统发育分析;基于叶绿体基因组高突变区构建特异性DNA条形码进行验证.结果:3 种败酱属植物叶绿体基因组呈四分体结构,全长分别为 159 585、158 919、158 919 bp,编码的基因数分别为 130、132、132,包含 8 个rRNA基因和 37 个tRNA基因,蛋白质编码基因数分别为 85、87、87.ccsA、ndhG、rpl23、ndhF序列作为特异性DNA条形码成功扩增 16 份样品,黄花败酱、白花败酱和少蕊败酱聚类在同一支,异叶败酱和糙叶败酱聚类在另一支.通过ccsA、ndhG、ndhF序列可进一步鉴别糙叶败酱和异叶败酱.结论:ccsA、ndhG、rpl23、ndhF序列可作为补充特异性DNA条形码区分败酱草及其近缘种.
Abstract
Objective:To develop specific DNA barcodes based on the chloroplast genome sequence analysis of the original plants of Patrinia medicinal materials to identify Patrniae Herba accurately and its related species.Methods:The chloroplast genomes of three species of the Patrinia plants,namely P.villosa,P.scabiosaefolia,and P.heterophylla,were sequenced using Illumina high-throughput sequencing technology.Bioinformatics software was employed for genome assembly,annotation,feature analysis,sequence comparison,and phylogenetic analysis.Specific DNA barcodes were constructed based on the highly variable regions of the chloroplast genome and were validated.Results:The chloroplast genomes of the three Patrinia plants exhibited a quadripartite structure,with total lengths of 159 585 bp,158 919 bp,and 158 919 bp,respectively.They encoded 130,132,and 132 genes,including 8 rRNA genes and 37 tRNA genes.The number of protein-coding genes was 85,87,and 87,respectively.The ccsA,ndhG,rpl23,and ndhF sequences could be used as specific DNA barcodes,successfully amplifying 16 samples.P.scabiosaefolia,P.villosa,and P.monandra clustered together in the same branch,while P.heterophylla and P.scabra clustered in another branch.The ccsA,ndhG,and ndhF sequences further distinguished P.heterophylla from P.scabra.Conclusion:The sequences of ccsA,ndhG,rpl23,and ndhF can be used as supplementary specific DNA barcodes to distinguish Patriniae Herba and its related species.
基金项目
国家药品监督管理局中药质量研究与评价重点实验室开放课题(AHYJ-KFKT-202103)
维普
万方数据