Diel transcriptional response of a California Current plankton microbiome to light, low iron, and enduring viral infection.

Phytoplankton and associated microbial communities provide organic carbon to oceanic food webs and drive ecosystem dynamics. However, capturing those dynamics is challenging. Here, an in situ, semi-Lagrangian, robotic sampler profiled pelagic microbes at 4 h intervals over ~2.6 days in North Pacific high-nutrient, low-chlorophyll waters. We report on the community structure and transcriptional dynamics of microbes in an operationally large size class (>5 µm) predominantly populated by dinoflagellates, ciliates, haptophytes, pelagophytes, diatoms, cyanobacteria (chiefly Synechococcus), prasinophytes (chiefly Ostreococcus), fungi, archaea, and proteobacteria. Apart from fungi and archaea, all groups exhibited 24-h periodicity in some transcripts, but larger portions of the transcriptome oscillated in phototrophs. Periodic photosynthesis-related transcripts exhibited a temporal cascade across the morning hours, conserved across diverse phototrophic lineages. Pronounced silica:nitrate drawdown, a high flavodoxin to ferredoxin transcript ratio, and elevated expression of other Fe-stress markers indicated Fe-limitation. Fe-stress markers peaked during a photoperiodically adaptive time window that could modulate phytoplankton response to seasonal Fe-limitation. Remarkably, we observed viruses that infect the majority of abundant taxa, often with total transcriptional activity synchronized with putative hosts. Taken together, these data reveal a microbial plankton community that is shaped by recycled production and tightly controlled by Fe-limitation and viral activity.

Widespread occurrence and genetic diversity of marine parasitoids belonging to Syndiniales (Alveolata).

Syndiniales are a parasitic order within the eukaryotic lineage Dinophyceae (Alveolata). Here, we analysed the taxonomy of this group using 43655 18S rRNA gene sequences obtained either from environmental data sets or cultures, including 6874 environmental sequences from this study derived from Atlantic and Mediterranean waters. A total of 5571 out of the 43655 sequences analysed fell within the Dinophyceae. Both bayesian and maximum likelihood phylogenies placed Syndiniales in five main groups (I-V), as a monophyletic lineage at the base of 'core' dinoflagellates (all Dinophyceae except Syndiniales), although the latter placement was not bootstrap supported. Thus, the two uncultured novel marine alveolate groups I and II, which have been highlighted previously, are confirmed to belong to the Syndiniales. These groups were the most diverse and highly represented in environmental studies. Within each, 8 and 44 clades were identified respectively. Co-evolutionary trends between parasitic Syndiniales and their putative hosts were not clear, suggesting they may be relatively 'general' parasitoids. Based on the overall distribution patterns of the Syndiniales-affiliated sequences, we propose that Syndiniales are exclusively marine. Interestingly, sequences belonging to groups II, III and V were largely retrieved from the photic zone, while Group I dominated samples from anoxic and suboxic ecosystems. Nevertheless, both groups I and II contained specific clades preferentially, or exclusively, retrieved from these latter ecosystems. Given the broad distribution of Syndiniales, our work indicates that parasitism may be a major force in ocean food webs, a force that is neglected in current conceptualizations of the marine carbon cycle.

In situ hybridization of Prochlorococcus and Synechococcus (marine cyanobacteria) spp. with RRNA-targeted peptide nucleic acid probes.

A simple method for whole-cell hybridization using fluorescently labeled rRNA-targeted peptide nucleic acid (PNA) probes was developed for use in marine cyanobacterial picoplankton. In contrast to established protocols, this method is capable of detecting rRNA in Prochlorococcus, the most abundant unicellular marine cyanobacterium. Because the method avoids the use of alcohol fixation, the chlorophyll content of Prochlorococcus cells is preserved, facilitating the identification of these cells in natural samples. PNA probe-conferred fluorescence was measured flow cytometrically and was always significantly higher than that of the negative control probe, with positive/negative ratio varying between 4 and 10, depending on strain and culture growth conditions. Prochlorococcus cells from open ocean samples were detectable with this method. RNase treatment reduced probe-conferred fluorescence to background levels, demonstrating that this signal was in fact related to the presence of rRNA. In another marine cyanobacterium, Synechococcus, in which both PNA and oligonucleotide probes can be used in whole-cell hybridizations, the magnitude of fluorescence from the former was fivefold higher than that from the latter, although the positive/negative ratio was comparable for both probes. In Synechococcus cells growing at a range of growth rates (and thus having different rRNA concentrations per cell), the PNA- and oligonucleotide-derived signals were highly correlated (r = 0.99). The chemical nature of PNA, the sensitivity of PNA-RNA binding to single-base-pair mismatches, and the preservation of cellular integrity by this method suggest that it may be useful for phylogenetic probing of whole cells in the natural environment.