Metabarcoding reveals potentially mixotrophic flagellates and picophytoplankton as key groups of phytoplankton in the Elbe estuary.

In estuaries, phytoplankton are faced with strong environmental forcing (e.g. high turbidity, salinity gradients). Taxa that appear under such conditions may play a critical role in maintaining food webs and biological carbon pumping, but knowledge about estuarine biota remains limited. This is also the case in the Elbe estuary where the lower 70 km of the water body are largely unexplored. In the present study, we investigated the phytoplankton composition in the Elbe estuary via metabarcoding. Our aim was to identify key taxa in the unmonitored reaches of this ecosystem and compare our results from the monitored area with available microscopy data. Phytoplankton communities followed distinct seasonal and spatial patterns. Community composition was similar across methods. Contributions of key classes and genera were correlated to each other (p < 0.05) when obtained from reads and biovolume (R2 = 0.59 and 0.33, respectively). Centric diatoms (e.g. Stephanodiscus) were the dominant group - comprising on average 55 % of the reads and 66-69 % of the biovolume. However, results from metabarcoding imply that microscopy underestimates the prevalence of picophytoplankton and flagellates with a potential for mixotrophy (e.g. cryptophytes). This might be due to their small size and sensitivity to fixation agents. We argue that mixotrophic flagellates are ecologically relevant in the mid to lower estuary, where, e.g., high turbidity render living conditions rather unfavorable, and skills such as phagotrophy provide fundamental advantages. Nevertheless, further findings - e.g. important taxa missing from the metabarcoding dataset - emphasize potential limitations of this method and quantitative biases can result from varying numbers of gene copies in different taxa. Further research should address these methodological issues but also shed light on the causal relationship of taxa with the environmental conditions, also with respect to active mixotrophic behavior.

Seasonality, rather than estuarine gradient or particle suspension/sinking dynamics, determines estuarine carbon distributions.

Estuaries are important components of the global carbon cycle; exchanging carbon between aquatic, atmospheric, and terrestrial environments, representing important loci for blue carbon storage and greenhouse gas emissions. However, how estuarine gradients affect sinking/suspended particles, and dissolved organic matter dynamic interactions remains unexplored. We fractionated suspended/sinking particles to assess and characterise carbon fate differences. We investigated bacterial colonisation (SYBR Green I) and exopolymer concentrations (TEP/CSP) with microscopy staining techniques. C/H/N and dry weight analysis identified particle composition differences. Meanwhile, nutrient and carbon analysis, and excitation and emission matrix evaluations with a subsequent parallel factor (PARAFAC) analysis characterised dissolved organic matter. The lack of clear salinity driven patterns in our study are presumably due to strong mixing forces and high particle heterogeneity along the estuary, with only density differences between suspended and sinking particles. Elbe estuary particles' organic portion is made up of marine-like (sinking) and terrestrial-like (suspended) signatures. Salinity did not have a significant role in microbial degradation and carbon composition, although brackish estuary portions were more biologically active. Indicative of increased degradation rates, leading to decreased greenhouse gas emissions, which are especially relevant for estuaries, with their disproportionate greenhouse gas emissions. Bacterial colonisation decreased seawards, indicative of decreased degradation, and shifts in microbial community composition and functions. Our findings span diverse strands of research, concerning steady carbon contributions from both marine and terrestrial sources, carbon aromaticity, humification index, and bioavailability. Their integration highlights the importance of the Elbe estuary as a model system, providing robust information for future policy decisions affecting dissolved and particulate matter dynamics within the Elbe Estuary.