Mixotrophs are increasingly recognized for their wide distribution in aquatic ecosystems and significant contributions to biogeochemical cycling. Many taxa within the phyla Chrysophyta, Cryptophyta, and Haptophyta are capable of phago-mixotrophy, however, phagotrophy in the Chlorophyta remains controversial due to insufficient research and solid evidence. In this study, we identified a new strain, Picochlorum sp. GLMF1 (Trebouxiophyceae), using 18S rRNA gene analysis and morphological observations. It displayed multi-cell division through autosporulation (two-or four-cell daughters) and has two unequal flagella that have never been reported in the genus Picochlorum. By using multiple methods, including 3D bioimaging analysis, acidic food vacuolelike compartment staining, and prey reduction calculation, we discovered and confirmed bacterivory in Picochlorum, which provided strong evidence for phago-mixotrophy in this green alga. In addition, we found that Picochlorum sp. GLMF1 cannot grow under complete darkness or prey-depleted conditions, suggesting that both light and bacteria are indispensable for this strain, and its mixotrophic nutrition mode is obligate. Like other phagophototrophs, Picochlorum sp. GLMF1 is capable of regulating their growth and ingestion rates according to light intensity and inorganic nutrient concentration. The confirmation of mixotrophy in this Picochlorum strain advances our understanding of the trophic roles of green algae, as well as the photosynthetic picoeukaryotes, in marine microbial food webs.

The silica frustules of diatom are regarded as an important physical defense against predation. While the biogenic silica content of diatoms is a major factor controlling the ingestion rate of copepods, how it affects the three‐dimensional structure and the mechanical properties of the frustules, and consequently copepod grazing, has not been studied. In this study, the diatoms Thalassiosira weissflogii and Amphora coffeaeformis were grown under different light intensities to manipulate their cellular biogenic silica contents. This manipulation generated diatom cells with distinct nano‐structures and mechanical strengths. The diatoms were then fed to the copepod Paracalanus pravus to test the effect of biogenic silica contents on copepod feeding. We demonstrated that the copepods preferred the low‐silica cells of T. weissflogii, but showed a preference for the high‐silica cells of A. coffeaeformis. Atomic force microscopy and X‐ray photoelectron spectroscopy analyses revealed that the chemical composition and mechanical properties of diatom frustules varied with silica contents. High silica containing T. weissflogii had higher frustule elasticity and hardness. In contrast, A. coffeaeformis containing more biogenic silica had lower frustule elasticity but silica level had little effect on the hardness of the frustule. Our results indicate that the nano‐structure and chemical properties of the frustules determine the mechanical strength of the diatom frustules. This nanoscale study improves our understanding of the relationship between diatom biogenic silica content and copepod grazing in marine planktonic food webs.

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