Environmental monitoring using next generation sequencing: rapid identification of macroinvertebrate bioindicator species

摘要: Invertebrate communities are central to many environmental monitoring programs. In freshwater ecosystems, aquatic macroinvertebrates collected, identified and then used infer ecosystem condition. Yet the key step of species identification is often not taken, as it requires a high level taxonomic expertise, which lacking in most organizations, or cannot be they morphologically cryptic represent little known groups. Identifying using DNA sequences can overcome these issues; with power next generation sequencing (NGS), for routine becomes feasible. this study, we test if NGS identify from field-collected samples an important bioindicator group, Chironomidae. We show that Cytochrome oxidase I (COI) B (CytB) provide accurate barcodes chironomid species. develop analysis pipeline identifying megablast searches quality generated 454 pyrosequencing against comprehensive reference libraries Sanger-sequenced voucher specimens. find COI successfully up 96% samples, but increased 99% when combined CytB sequences. Accurate depends on having at least five species; below expected were detected. Incorrect incorporation some multiplex identifiers (MID’s) tag was likely cause, errors could detected MID tags forward reverse primers. also found strong quantitative relationship between number individuals showing may possible estimate abundance data. Next two genes successful However, detecting data sets critical include control establish thresholds The approach developed here lead species-level diagnostic ecosystems.