Carew Melissa E, Coleman Rhys A, Hoffmann Ary A
School of BioSciences, The University of Melbourne, Melbourne, Victoria, Australia.
Applied Research, Melbourne Water Corporation, Melbourne, Victoria, Australia.
PeerJ. 2018 Jun 13;6:e4980. doi: 10.7717/peerj.4980. eCollection 2018.
High throughput DNA sequencing of bulk invertebrate samples or metabarcoding is becoming increasingly used to provide profiles of biological communities for environmental monitoring. As metabarcoding becomes more widely applied, new reference DNA barcodes linked to individual specimens identified by taxonomists are needed. This can be achieved through using DNA extraction methods that are not only suitable for metabarcoding but also for building reference DNA barcode libraries.
In this study, we test the suitability of a rapid non-destructive DNA extraction method for metabarcoding of freshwater invertebrate samples.
This method resulted in detection of taxa from many taxonomic groups, comparable to results obtained with two other tissue-based extraction methods. Most taxa could also be successfully used for subsequent individual-based DNA barcoding and taxonomic identification. The method was successfully applied to field-collected invertebrate samples stored for taxonomic studies in 70% ethanol at room temperature, a commonly used storage method for freshwater samples.
With further refinement and testing, non-destructive extraction has the potential to rapidly characterise species biodiversity in invertebrate samples, while preserving specimens for taxonomic investigation.
对大量无脊椎动物样本进行高通量DNA测序或代谢条形码分析正越来越多地用于为环境监测提供生物群落概况。随着代谢条形码分析的应用越来越广泛,需要与分类学家鉴定的单个标本相关联的新的参考DNA条形码。这可以通过使用不仅适用于代谢条形码分析而且适用于构建参考DNA条形码文库的DNA提取方法来实现。
在本研究中,我们测试了一种快速无损DNA提取方法对淡水无脊椎动物样本进行代谢条形码分析的适用性。
该方法检测到了许多分类群中的分类单元,与另外两种基于组织的提取方法所获得的结果相当。大多数分类单元也可以成功用于后续基于个体的DNA条形码分析和分类鉴定。该方法已成功应用于室温下保存在70%乙醇中用于分类学研究的野外采集的无脊椎动物样本,这是淡水样本常用的保存方法。
经过进一步改进和测试,无损提取有可能快速表征无脊椎动物样本中的物种生物多样性,同时保留标本用于分类学研究。