Metabarcoding vs Microscopy: Comparison of Methods To Monitor Phytoplankton Communities

• DOI: 10.1021/acsestwater.3c00176
• Publication Date: 2023-07-21T14:17:56Z
• Authors: Agneta Andersson; Li Zhao; Sonia Brugel; Daniela Figueroa; Siv Huseby
• Keywords: Oceanography, Hydrology and Water Resources; group/class-level consistency; Sampling volume; 14. Life underwater; carbon biomass; Oceanografi, hydrologi och vattenresurser; gene abundance; 6. Clean water; diversity

Phytoplankton are used worldwide to monitor environmental status in aquatic systems. Long-time series of microscopy-analyzed phytoplankton are available from many monitoring stations. The microscopy-method is however time consuming and has short-comings. DNA metabarcoding has been suggested as an alternative method, but the consistency between different methods need further investigation. We performed a comparative study of microscopy and metabarcoding analyzing micro- and nanophytoplankton. For metabarcoding, 25-1000 ml seawater were filtered, DNA extracted and the 18S and 16S rRNA gene amplicons sequenced. For microscopy, based on the Utermöhl method we evaluated the use of three metrics: abundance, biovolume and carbon biomass. At the genus, species, and unidentified taxa level, metabarcoding generally showed higher taxonomic diversity than microscopy, and diversity was already captured at the lowest filtration volume tested, 25 ml. Metabarcoding and microscopy displayed relatively similar distribution pattern at the group level. The results showed that the relative abundances of the 18S rRNA amplicon at the group level best fitted the microscopy carbon biomass metric. The results are promising for implementing DNA metabarcoding as a complement to microscopy in phytoplankton monitoring, especially if databases would be improved and group level indexes could be applied to classify the environmental state of water bodies. Originally included in thesis in manuscript form.