Small photosynthetic eukaryotes are key primary producers in marine waters. In recent years, their diversity has been studied by the analysis of 18S rRNA gene sequences directly amplified and cloned from filtered natural samples. However, these clone libraries are often dominated by nonphotosynthetic organisms and few sequences from autotrophs are recovered. In the present paper, we developed a new approach based on flow cytometry. Photosynthetic pico-, nano- and phycoerythrin-containing (PE-) eukaryotes from the coastal English Channel were sorted based on their size and pigment fluorescence. 18S rRNA gene libraries were constructed from the DNA of sorted cells. We addressed methodological issues linked to the relatively low concentration of these cells. This novel approach confirmed that, in the English Channel, pico-eukaryotes are dominated by three genera Micromonas, Ostreococcus and Bathycoccus, while PE-eukaryotes are mainly cryptophytes from clade 4. It also revealed that nano-eukaryotes are dominated by haptophytes with important contributions from small diatoms and Prasinophyceae. It should be emphasized that haptophytes were nearly absent from clone libraries constructed from filtered samples, which explains why they have been overlooked in previous studies. The new strategy should be very useful to conduct similar studies on other specific populations that can be discriminated by flow cytometry (e.g. red tide organisms or uncultivated protists).
The genus Micromonas comprises phytoplankton that show among the widest latitudinal distributions on Earth, and members of this genus are recurrently infected by prasinoviruses in contrasted thermal ecosystems. In this study, we assessed how temperature influences the interplay between the main genetic clades of this prominent microalga and their viruses. The growth of three Micromonas strains (Mic-A, Mic-B, Mic-C) and the stability of their respective lytic viruses (MicV-A, MicV-B, MicV-C) were measured over a thermal range of 4-32.5°C. Similar growth temperature optima (T opt ) were predicted for all three hosts but Mic-B exhibited a broader thermal tolerance than Mic-A and Mic-C, suggesting distinct thermoacclimation strategies. Similarly, the MicV-C virus displayed a remarkable thermal stability compared with MicV-A and MicV-B. Despite these divergences, infection dynamics showed that temperatures below T opt lengthened lytic cycle kinetics and reduced viral yield and, notably, that infection at temperatures above T opt did not usually result in cell lysis. Two mechanisms operated depending on the temperature and the biological system. Hosts either prevented the production of viral progeny or maintained their ability to produce virions with no apparent cell lysis, pointing to a possible switch in the viral life strategy. Hence, temperature changes critically affect the outcome of Micromonas infection and have implications for ocean biogeochemistry and evolution.
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