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doi: 10.1038/ismej.2012.85. Epub 2012 Jul 19.

Taming the smallest predators of the oceans

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Taming the smallest predators of the oceans

Javier del Campo et al. ISME J.2013 Feb.

Abstract

Protists (unicellular eukaryotes) arguably account for most eukaryotic diversity and are central players of the biosphere. Known protist diversity and biology is largely based on cultured strains. Yet, environmental molecular surveys have unveiled entirely novel lineages that, as their prokaryotic counterparts, are essentially uncultured. Culture bias is an important drawback for any microbe-related science and is particularly severe for heterotrophic protists, which depend on organic food sources for growth. Here, we show how ecologically significant bacterivorous protists have been brought into culture by mimicking in situ conditions. Single cells sorted by serial dilution or flow cytometry were inoculated into seawater amended with natural bacterial assemblage at nearly in situ abundances. Strains belonging to lineages only known so far from environmental sequencing were isolated. Among them, Minorisa minuta gen. nov. sp. nov. forms a novel branch within Rhizaria, holding a key evolutionary position, and with an average size of 1.4 μm represents one of the smallest bacterial grazers known to date. It has a worldwide planktonic distribution and can account for 5% of heterotrophic protists communities in coastal waters. Physiological features of this strain can partly explain its success in the environment. Culturing ecologically relevant but elusive protists provide invaluable material for ecophysiology, genomics, ecosystem modeling and evolutionary issues.

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Figures

Figure 1
Figure 1
(a) ML phylogenetic tree with complete 18S rDNA sequences showing the position ofMinorisa minuta within the Cercozoa. The scale bar indicates 0.1 substitutions per position. The coverage of the specific probe CRN02 is shown in grey. (b) SEM image of aMinorisa minuta cell of 1.6 μm in size and possessing a single flagellum. (c) Growth ofMinorisa minuta with natural bacteria as prey in a batch culture.
Figure 2
Figure 2
(a) Abundances ofMinorisa minuta cells at various sampling sites as estimated by TSA-FISH counts. (b) Abundances ofMinorisa minuta cells during a temporal study at Blanes Bay (year 2007), together with the abundance of heterotrophic flagellates, estimated by epifluorescence (log scale). Bars represent the contribution (in %) ofMinorisa minuta cells to heterotrophic flagellates.
Figure 3
Figure 3
(a) Correlation between the abundances ofMinorisa minuta and natural bacteria during the seasonal study (year 2007) at Blanes Bay. (b) Cell size distribution ofMinorisa minuta in these samples (1148 cells measured by TSA-FISH).
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