中药材2025，Vol.48Issue(12)：2993-2997,5.DOI:10.13863/j.issn1001-4454.2025.12.009
基于地鳖线粒体基因组差异鉴定土鳖虫的性别
Gender Identification of Eupolyphaga Steleophaga Based on Mitochondrial Genome Differences of Eupolyphaga sinensis
摘要
Abstract
Objective:To establish a rapid and accurate molecular gender identification method for the mixed use of female and male Eudolyphaca sinensis individuals in the medicinal materials of Eupolyphaga Steleophaga,providing a technical support for the qual-ity control of the medicinal materials.Methods:The complete mitochondrial genome sequences of adult female and male Eupolyphaga sinensis were determined respectively by using Illumina high-throughput sequencing technology.The mitochondrial genomic difference loci between female and male individuals were compared and analyzed,and sequencing verification and enzyme digestion site analysis were conducted.Based on specific enzyme digestion sites,a PCR-RFLP molecular identification method was established.Results:Three mitochondrial genome sequences of female and male Eupolyphaga sinensis were successfully obtained,with a total length of 15 551 bp.There were three stable base difference sites(sites 4 113,4 413,8 262)between female and male individuals.Two pairs of universal primers designed based on the fragment where the difference site was located could effectively amplify the target sequence.Further,the PCR-RFLP identification system was successfully established by using the EcoT14 I enzyme digestion site.This method could complete the gender identification of Eupolyphaga sinensis samples within 2.5 hours.Conclusion:The PCR-RFLP gender identification method established in this study is highly accurate,convenient and fast,providing feasible technical solution for the quality control and standard formulation of the gender of Eupolyphaga Steleophaga medicinal materials.关键词
土鳖虫/地鳖/线粒体基因组/性别鉴定/聚合酶链式反应-限制性片段长度多态性Key words
Eupolyphaga Steleophaga/Eupolyphaga sinensis Walker/Mitochondrial genome/Gender identification/Polymerase chain reaction-restriction fragment length polymorphism分类
医药卫生引用本文复制引用
邹何元,梁莹莹,吴文如,江珮瑜,张雨薇,来慧丽..基于地鳖线粒体基因组差异鉴定土鳖虫的性别[J].中药材,2025,48(12):2993-2997,5.基金项目
国家中医药管理局张丹雁全国老药工传承工作室建设项目(国中医药人教函[2024]255号) （国中医药人教函[2024]255号）
2024年广东食品药品职业学院科研项目(2024ZR27) （2024ZR27）
广州中医药大学大学生创新创业训练计划项目(202410572308) （202410572308）
2024年广东省研究生教育创新计划项目(2024ANLK_018) （2024ANLK_018）
