Viral ecology of organic and inorganic particles in aquatic systems: avenues for further research
Viral abundance and processes in the water column and sediments are well studied for some systems; however, we know relatively little about virus-host interactions on particles and how particles influence these interactions. Here we review virus-prokaryote interactions on inorganic and organic particles in the water column. Profiting from recent methodological progress, we show that confocal laser scanning microscopy in combination with lectin and nucleic acid staining is one of the most powerful methods to visualize the distribution of viruses and their hosts on particles such as organic aggregates. Viral abundance on suspended matter ranges from 10 to 10 ml. The main factors controlling viral abundance are the quality, size and age of aggregates and the exposure time of viruses to aggregates. Other factors such as water residence time likely act indirectly. Overall, aggregates appear to play a role of viral scavengers or reservoirs rather than viral factories. Adsorption of viruses to organic aggregates or inorganic particles can stimulate growth of the free-living prokaryotic community, e.g. by reducing viral lysis. Such mechanisms can affect microbial diversity, food web structure and biogeochemical cycles. Viral lysis of bacterio- and phytoplankton influences the formation and fate of aggregates and can, for example, result in a higher stability of algal flocs. Thus, viruses also influence carbon export; however, it is still not clear whether they short-circuit or prime the biological pump. Throughout this review, emphasis has been placed on defining general problems and knowledge gaps in virus-particle interactions and on providing avenues for further research, particularly those linked to global change.
Effects of suspended matter quality and virus abundance on microbial parameters: experimental evidence from a large European river
In riverine water, both suspended particulate material and viruses are prominent ecological factors. The existence of various particle types and differences in viral abundance impose variability in microenvironments. Particulates and their microbial surrounding may interact in several ways, this interaction being strongly dependent on particle quality and the abundance of organisms involved. In laboratory experiments, we used different suspended matter types (fresh and aged mineral sediment and leaf litter, river snow) that typically occur in riverine environments as model particles. We investigated the effects of particle quality and different ambient viral abundances (×1, ×2 enrichments, and inactivated viruses) on several microbial parameters (changes in bacterial and viral abundances, bacterial production, specific bacterial production) of both the free-living and particle-attached fractions using water from a floodplain system of the Danube River (Austria). Both seston quality and variable viral abundances in the bulk water influenced some microbial parameters. The average abundance of bacteria and viruses was significantly higher on organic than on inorganic particles and on aged particles (for both sediment and leaf litter). Changes in bacterial abundance during the course of the experiments were also influenced by particle quality, with, for example, aged sediment favoring increasing abundances. Virus:bacterium ratios (VBR) were significantly higher on organic than on inorganic particles, but significantly lower on suspended particles than in the plank-tonic fraction. Typically, bacterial secondary production (overall and cell-specific) was higher on particles than in bulk water. Bacterial productivity in the ambient water was negatively affected by the abundance of planktonic viruses but positively affected by that of attached viruses. These findings from experimental systems may foster studies of particle-rich environments.
Deciphering diatom biochemical pathways via whole-cell proteomics
Diatoms play a critical role in the oceans' carbon and silicon cycles; however, a mechanistic understanding of the biochemical processes that contribute to their ecological success remains elusive. Completion of the Thalassiosira pseudonana genome provided 'blueprints' for the potential biochemical machinery of diatoms, but offers only a limited insight into their biology under various environmental conditions. Using high-throughput shotgun proteomics, we identified a total of 1928 proteins expressed by T. pseudonana cultured under optimal growth conditions, enabling us to analyze this diatom's primary metabolic and biosynthetic pathways. Of the proteins identified, 70% are involved in cellular metabolism, while 11% are involved in the transport of molecules. We identified all of the enzymes involved in the urea cycle, thereby describing the complete pathway to convert ammonia to urea, along with urea transporters, and the urea-degrading enzyme urease. Although metabolic exchange between these pathways remains ambiguous, their constitutive presence suggests complex intracellular nitrogen recycling. In addition, all C(4) related enzymes for carbon fixation have been identified to be in abundance, with high protein sequence coverage. Quantification of mass spectra acquisitions demonstrated that the 20 most abundant proteins included an unexpectedly high expression of clathrin, which is the primary structural protein involved in endocytic transport. This result highlights a previously overlooked mechanism for the inter- and intra-cellular transport of nutrients and macromolecules in diatoms, potentially providing a missing link to organelle communication and metabolite exchange. Our results demonstrate the power of proteomics, and lay the groundwork for future comparative proteomic studies and directed analyses of specifically expressed proteins and biochemical pathways of oceanic diatoms.
Role of dissolved nitrate and phosphate in isolates of and toxin-producing
, a producer of toxins associated with diarrhetic shellfish poisoning (DSP) and/or pectenotoxins (PTXs), is a mixotrophic species that requires both ciliate prey and light for growth. Linkages have been described in the literature between natural abundances of the predator and its prey, , and culture experiments have demonstrated that prey, in addition to light, is required for toxin production by ; together these suggest is a critical component for growth and toxicity. However, little is known about the role of dissolved inorganic nutrients on growth or that of toxin-producing . Accordingly, a series of experiments were conducted to investigate the possible uptake of dissolved nitrate and phosphate by 1) starved of prey, 2) feeding on , and 3) grown in nutritionally-modified media. All single-clone or mixed cultures were monitored for dissolved and particulate nutrient levels over the growth cycle, as well as growth rate, biomass, and toxin production when appropriate. did not utilize dissolved nitrate or phosphate in the medium under any nutrient regime tested, i.e., nutrient-enriched and nutrient-reduced, in the absence or presence of prey, or during any growth phase monitored, i.e., exponential and plateau phases. Changes in particulate phosphorus and nitrogen in , were instead, strongly influenced by the consumption of prey, and these levels quickly stabilized once prey were no longer available. , on the other hand, rapidly assimilated dissolved nitrate and phosphate into its particulate nutrient fraction, with maximum uptake rates of 1.38 pmol N/cell/day and 1.63 pmol P/cell/day. While did not benefit directly from the dissolved nitrate and phosphate, its growth (0.37±0.01 day) and toxin production rates for okadaic acid (OA), dinophysistoxin-1 (DTX1) or pectenotoxin-2 (PTX2), 0.1, 0.9 and 2.6 pg /cell/day, respectively, were directly coupled to prey availability. These results suggest that while dissolved nitrate and phosphate do not have a direct effect on toxin production or retention by , these nutrient pools contribute to prey growth and biomass, thereby indirectly influencing blooms and overall toxin in the system.