甘肃中医药大学学报2026，Vol.43Issue(1)：41-48,8.DOI:10.16841/j.issn1003-8450.2026.01.06
基于DNA条形码鉴别半夏及其混伪品
DNA barcode-based authentication of Pinellia ternata and its adulterants
摘要
Abstract
Objective To establish an accurate identification method for Pinellia ternata and its common adul-terants based on the internal transcribed spacer 2(ITS2)sequence and its secondary structure.Methods 19 bat-ches of samples,including Pinellia ternata,Pinellia pedatisecta,Pinellia cordata,Typhonium flagelliforme,and Ari-saema heterophyllum,were collected.Genomic DNA was extracted,amplified with universal ITS primers,and bidi-rectionally sequenced.11 ITS2 sequences of related species were retrieved from NCBI.Self-generated and database sequences were integrated to calculate intra-and inter-species Kimura 2-parameter(K2P)genetic distances using MEGA 11.0,followed by Neighbor-Joining(NJ)tree construction.ITS2 secondary structures were predicted via online tools.Results Intra-specific genetic distances ranged from 0-0.011,while inter-specific distances spanned 0.055-0.339,demonstrating distinct barcoding gaps.Within the studied taxa,intra-generic genetic distances consis-tently measured below 0.150,whereas inter-generic distances uniformly exceeded 0.150,supporting 0.150 as an em-pirical genus-level threshold.The NJ tree revealed monophyletic clustering for all species except Typhonium gigan-teum.and Typhonium flagelliforme,which failed to form a cohesive clade,and Arisaema heterophyllum,which branched independently,indicating taxonomic inconsistencies.ITS2 secondary structures exhibited a conserved"central loop with four helical arms"across species except Arisaema heterophyllum,with genus-specific variations in loop morphology and helix arrangements enabling reliable differentiation of closely related taxa.Conclusion ITS2 sequence coupled with secondary structure analysis provides a robust tool for authenticating Pinellia ternata and its adulterants while exposing phylogenetic incongruences in traditional classification.This approach is recommended as a complementary DNA barcode for quality control of Pinellia-based medicinal products.关键词
半夏/DNA条形码/内转录间隔区序列/二级结构/分子鉴定/混伪品Key words
Pinellia ternata/DNA barcoding/ITS2 sequence/secondary structure/molecular authentication/adulterants分类
医药卫生引用本文复制引用
孙一帆,徐义菁,郭雯雅,吴亚楠,王欢,王孟虎,魏忠侠,张露平..基于DNA条形码鉴别半夏及其混伪品[J].甘肃中医药大学学报,2026,43(1):41-48,8.基金项目
安徽省高等学校科学研究项目(2023AH053216) （2023AH053216）
安徽省高校优秀拔尖人才培育资助项目(YQYB2024085) （YQYB2024085）
亳州市重点研发计划项目(bzzc2023030,bzzc2024057) （bzzc2023030,bzzc2024057）
亳州学院质量工程项目(2024XJXM041). （2024XJXM041）
