Abstract

AbstractEnvironmental molecular sequencing has revealed an abundance of microorganisms that were previously unknown, mainly because most had not been cultured in the laboratory. Within this novel diversity, there are the uncultured MAST clades (MArine STramenopiles), which are major components of marine heterotrophic flagellates (HFs) thought to be active bacterial grazers. In this study, we investigated the gene expression of natural HFs in a mixed community where bacterivory was promoted. Using fluorescence in situ hybridization and 18S rDNA derived from metatranscriptomics, we followed the taxonomic dynamics during the incubation, and confirmed the increase in relative abundance of different MAST lineages. We then used single cell genomes of several MAST species to gain an insight into their most expressed genes, with a particular focus on genes related to phagocytosis. The genomes of MAST‐4A and MAST‐4B were the most represented in the metatranscriptomes, and we identified highly expressed genes of these two species involved in motility and cytoskeleton remodeling, as well as many lysosomal enzymes. Particularly relevant were the cathepsins, which are characteristic digestive enzymes of the phagolysosome and the rhodopsins, perhaps used for vacuole acidification. The combination of single cell genomics and metatranscriptomics gives insights on the phagocytic capacity of uncultured and ecologically relevant HF species.

Highlights

  • Heterotrophic flagellates (HFs) are widespread throughout the eukaryotic tree of life and may represent the most ancient eukaryote lifestyle (Cavalier-Smith 2006; Jürgens and Massana 2008; Adl et al 2019)

  • Growth of marine heterotrophic flagellates (HFs) in an unamended incubation We examined a mixed community of picoplanktonic microbes

  • Using direct epifluorescence microscopy counts, we evaluated the temporal dynamics of several components of this mixed assemblage (Fig. 1a)

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Summary

Introduction

Heterotrophic flagellates (HFs) are widespread throughout the eukaryotic tree of life and may represent the most ancient eukaryote lifestyle (Cavalier-Smith 2006; Jürgens and Massana 2008; Adl et al 2019) These colorless flagellated protists are important consumers of primary and secondary production in marine ecosystems (Arndt et al 2000; Worden et al 2015), and play a pivotal role in microbial food webs by ensuring the recycling of nutrients. They make a significant contribution, it is difficult to assess their diversity because they cannot be differentiated by microscopy and most remain uncultured. A mature phagolysosome is characterized by the presence of a range of lysosomal acid hydrolases such as proteases, lysozymes, S149

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