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引用本文:陈金萍,周春花,欧阳珊,黄晓晨,吴小平.鄱阳湖流域蚌类环境DNA宏条形码引物的筛选验证.湖泊科学,2021,33(4):1254-1264. DOI:10.18307/2021.0425
Chen Jinping,Zhou Chunhua,Ouyang Shan,Huang Xiaochen,Wu Xiaoping.Universal primers screening and verification for environmental DNA metabarcoding of freshwater mussels in the Lake Poyang Basin. J. Lake Sci.2021,33(4):1254-1264. DOI:10.18307/2021.0425
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鄱阳湖流域蚌类环境DNA宏条形码引物的筛选验证
陈金萍, 周春花, 欧阳珊, 黄晓晨, 吴小平
南昌大学生命科学学院, 南昌 330031
摘要:
环境DNA技术是一种非侵入、高灵敏、高效率且对环境无破坏性,对生物体无损伤的调查工具.为了筛选适合于蚌类环境DNA生物多样性研究的宏条形码引物,本研究通过对鄱阳湖流域24种常见蚌的基因组DNA进行普通扩增和高通量测序筛选了11对引物(设计了9对通用引物及从相关文献中引用了2对引物),结果显示引物cyt b和16S rRNA具有良好的扩增效果和高辨别度.进一步用环境样本(n=6)并结合传统采样技术对这2对引物进行验证,结果表明:使用引物16S rRNA共注释到蚌科物种6属8种,蚌科物种序列占总序列数的26.69%;而使用引物cyt b共注释到蚌科物种4属6种,蚌科物种序列占总序列数的6.60%.引物16S rRNA更适合用于蚌类环境DNA生物多样性研究的宏条形码引物.在生物多样性监测中环境DNA技术可以作为传统方法的有效补充,且同时使用多对引物,可增加可信度和检测率.
关键词:  环境DNA  生物多样性  蚌类  分子标记  鄱阳湖流域
DOI:10.18307/2021.0425
分类号:
基金项目:国家重点研发计划“蓝色粮仓”重点专项(2018YFD0900801)资助.
Universal primers screening and verification for environmental DNA metabarcoding of freshwater mussels in the Lake Poyang Basin
Chen Jinping, Zhou Chunhua, Ouyang Shan, Huang Xiaochen, Wu Xiaoping
College of Life Science, Nanchang University, Nanchang 330031, P. R. China
Abstract:
Environmental DNA (eDNA) technology is a non-invasive, highly sensitive, efficient and non-destructive to the environment, no damage to the organism investigation tool. To select suitable metabarcoding primers for freshwater mussel's environmental DNA biodiversity research, 11 pairs of primers (with 9 pairs of universal primers designed and 2 pairs cited from relevant literature) were screened by general amplification and high-throughput sequencing of genomic DNA of 24 common mussels in the Lake Poyang Basin. The results showed that the primer cyt b and 16S rRNA had good amplification effect and high discrimination. The two pairs of primers were further verified with environmental samples (n=6) and traditional sampling. It was found that 8 mussels species belonging to 6 genera were annotated by 16S rRNA which accounted for 26.69% of the total sequences, and 5 mussels species belonging to 4 genera were annotated by cyt b which accounted for 6.60% of the total sequences. The primer 16S rRNA was more suitable used for metabarcoding primer for the research of environmental DNA biodiversity in freshwater mussels. Environmental DNA technology can be used as an effective supplement to traditional methods in biodiversity monitoring. The reliability and detection rate can be increased by using multiple pairs of primers at the same time.
Key words:  Environmental DNA  biodiversity  freshwater mussel  molecular markers  Lake Poyang Basin
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