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  1. Abstract Phago-mixotrophy, the combination of photoautotrophy and phagotrophy in mixoplankton, organisms that can combine both trophic strategies, have gained increasing attention over the past decade. It is now recognized that a substantial number of protistan plankton species engage in phago-mixotrophy to obtain nutrients for growth and reproduction under a range of environmental conditions. Unfortunately, our current understanding of mixoplankton in aquatic systems significantly lags behind our understanding of zooplankton and phytoplankton, limiting our ability to fully comprehend the role of mixoplankton (and phago-mixotrophy) in the plankton food web and biogeochemical cycling. Here, we put forward five research directions that we believe will lead to major advancement in the field: (i) evolution: understanding mixotrophy in the context of the evolutionary transition from phagotrophy to photoautotrophy; (ii) traits and trade-offs: identifying the key traits and trade-offs constraining mixotrophic metabolisms; (iii) biogeography: large-scale patterns of mixoplankton distribution; (iv) biogeochemistry and trophic transfer: understanding mixoplankton as conduits of nutrients and energy; and (v) in situ methods: improving the identification of in situ mixoplankton and their phago-mixotrophic activity. 
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  2. Variability in community composition is often attributed to underlying differences in physical environments. However, predator–prey interactions can play an equally important role in structuring communities. Although environmental differences select for different species assemblages, less is known about their impacts on trait compositions. We develop a trait-based analysis of plankton communities of the southern California Current System across multiple trophic levels, from bacteria to mesozooplankton, and over a gradient of environmental conditions, from the oligotrophic open ocean to coastal upwelling. Across a factor of four differences in total community biomass, we observe similarities in the size structure along the environmental gradient, with the most pronounced departures from proportional variations in the biomasses found in the largest protists (> 40 lm). Differences in the trait distributions emerge within a small range of size groups: greater biomass contribution of larger autotrophs (> 10 lm) is observed only for the upwelling region. 
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