Arsenate Resistance in the Unicellular Marine Diazotroph Crocosphaera watsonii

The toxic arsenate ion can behave as a phosphate analog, and this can result in arsenate toxicity especially in areas with elevated arsenate to phosphate ratios like the surface waters of the ocean gyres. In these systems, cellular arsenate resistance strategies would allow phytoplankton to ameliorate the effects of arsenate transport into the cell. Despite the potential coupling between arsenate and phosphate cycling in oligotrophic marine waters, relatively little is known about arsenate resistance in the nitrogen-fixing marine cyanobacteria that are key components of the microbial community in low nutrient systems. The unicellular diazotroph, Crocosphaera watsonii WH8501, was able to grow at reduced rates with arsenate additions up to 30 nM, and estimated arsenate to phosphate ratios of 6:1. The genome of strain WH8501 contains homologs for arsA, arsH, arsB, and arsC, allowing for the reduction of arsenate to arsenite and the pumping of arsenite out of the cell. The short-term addition of arsenate to the growth medium had no effect on nitrogen fixation. However, arsenate addition did result in the up-regulation of the arsB gene with increasing arsenate concentrations, indicating the induction of the arsenate detoxification response. The arsB gene was also up-regulated by phosphorus stress in concert with a gene encoding the high-affinity phosphate binding protein pstS. Both genes were down-regulated when phosphate was re-fed to phosphorus-stressed cells. A field survey of surface water from the low phosphate western North Atlantic detected expression of C. watsonii arsB, suggestive of the potential importance of arsenate resistance strategies in this and perhaps other systems

Arsenate resistance in the unicellular marine diazotroph Crocosphaera watsonii

© The Author(s), 2011. This is an open-access article subject to a non-exclusive license between the authors and Frontiers Media SA, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited. The definitive version was published in Frontiers in Microbiology 2 (2011): 214, doi:10.3389/fmicb.2011.00214.The toxic arsenate ion can behave as a phosphate analog, and this can result in arsenate toxicity especially in areas with elevated arsenate to phosphate ratios like the surface waters of the ocean gyres. In these systems, cellular arsenate resistance strategies would allow phytoplankton to ameliorate the effects of arsenate transport into the cell. Despite the potential coupling between arsenate and phosphate cycling in oligotrophic marine waters, relatively little is known about arsenate resistance in the nitrogen-fixing marine cyanobacteria that are key components of the microbial community in low nutrient systems. The unicellular diazotroph, Crocosphaera watsonii WH8501, was able to grow at reduced rates with arsenate additions up to 30 nM, and estimated arsenate to phosphate ratios of 6:1. The genome of strain WH8501 contains homologs for arsA, arsH, arsB, and arsC, allowing for the reduction of arsenate to arsenite and the pumping of arsenite out of the cell. The short-term addition of arsenate to the growth medium had no effect on nitrogen fixation. However, arsenate addition did result in the up-regulation of the arsB gene with increasing arsenate concentrations, indicating the induction of the arsenate detoxification response. The arsB gene was also up-regulated by phosphorus stress in concert with a gene encoding the high-affinity phosphate binding protein pstS. Both genes were down-regulated when phosphate was re-fed to phosphorus-stressed cells. A field survey of surface water from the low phosphate western North Atlantic detected expression of C. watsonii arsB, suggestive of the potential importance of arsenate resistance strategies in this and perhaps other systems.This research was funded in part by the National Science Foundation #OCE-0451419, and the Center for Microbial Oceanography: Research and Education

Alkaline phosphatase activity and regulation in the North Pacific Subtropical Gyre

Alkaline phosphatase (AP) activity (APA) was measured at several stations in the North Pacific Subtropical Gyre in July 2008, and in a series of nutrient addition experiments: nitrate plus ammonium (+N) or phosphate (+P), to study APA regulation and to evaluate the capacity of picoplankton organisms (i.e., in the 0.2–2‐µm size range) to access the AP‐hydrolyzable fraction of dissolved organic phosphorus (DOP). The data indicated a primary limitation of the biomass by nitrogen. Both total (measured with a soluble DOP analog) and cell‐specific (measured with the enzyme‐labeled fluorescence [ELF] phosphate cell labeling method) APA were enhanced in the +N samples and reduced in the +P samples, suggesting that DOP is an important resource for picoplankton nutrition. Cell‐free APA represented > 65% of the APA in all samples, but its contribution to total APA significantly decreased in the +N treatment as microbial biomass increased. In the +N treatment, < 5% and up to 96% of the cells in the heterotrophic bacteria‐enriched and picophytoplankton‐enriched fractions, espectively, were ELF‐alcohol‐labeled after 5 d. Following N enrichment, the microbial assemblage shifted from cell‐free phosphatase dominated under N limitation and P stress (i.e., physiological response) to picophytoplankton‐based phosphatase dominated under P limitation (i.e., production or growth rate limitation). If, as predicted, the ocean evolves towards P limitation, DOP availability would become of major importance to sustain productivity

Niche of harmful alga Aureococcus anophagefferens revealed through ecogenomics

Harmful algal blooms (HABs) cause significant economic and ecological damage worldwide. Despite considerable efforts, a comprehensive understanding of the factors that promote these blooms has been lacking, because the biochemical pathways that facilitate their dominance relative to other phytoplankton within specific environments have not been identified. Here, biogeochemical measurements showed that the harmful alga Aureococcus anophagefferens outcompeted co-occurring phytoplankton in estuaries with elevated levels of dissolved organic matter and turbidity and low levels of dissolved inorganic nitrogen. We subsequently sequenced the genome of A. anophagefferens and compared its gene complement with those of six competing phytoplankton species identified through metaproteomics. Using an ecogenomic approach, we specifically focused on gene sets that may facilitate dominance within the environmental conditions present during blooms. A. anophagefferens possesses a larger genome (56 Mbp) and has more genes involved in light harvesting, organic carbon and nitrogen use, and encoding selenium- and metal-requiring enzymes than competing phytoplankton. Genes for the synthesis of microbial deterrents likely permit the proliferation of this species, with reduced mortality losses during blooms. Collectively, these findings suggest that anthropogenic activities resulting in elevated levels of turbidity, organic matter, and metals have opened a niche within coastal ecosystems that ideally suits the unique genetic capacity of A. anophagefferens and thus, has facilitated the proliferation of this and potentially other HABs

Metatranscriptome analyses indicate resource partitioning between diatoms in the field

Diverse communities of marine phytoplankton carry out half of global primary production. The vast diversity of the phytoplankton has long perplexed ecologists because these organisms coexist in an isotropic environment while competing for the same basic resources (e.g., inorganic nutrients). Differential niche partitioning of resources is one hypothesis to explain this “paradox of the plankton,” but it is difficult to quantify and track variation in phytoplankton metabolism in situ. Here, we use quantitative metatranscriptome analyses to examine pathways of nitrogen (N) and phosphorus (P) metabolism in diatoms that cooccur regularly in an estuary on the east coast of the United States (Narragansett Bay). Expression of known N and P metabolic pathways varied between diatoms, indicating apparent differences in resource utilization capacity that may prevent direct competition. Nutrient amendment incubations skewed N/P ratios, elucidating nutrient-responsive patterns of expression and facilitating a quantitative comparison between diatoms. The resource-responsive (RR) gene sets deviated in composition from the metabolic profile of the organism, being enriched in genes associated with N and P metabolism. Expression of the RR gene set varied over time and differed significantly between diatoms, resulting in opposite transcriptional responses to the same environment. Apparent differences in metabolic capacity and the expression of that capacity in the environment suggest that diatom-specific resource partitioning was occurring in Narragansett Bay. This high-resolution approach highlights the molecular underpinnings of diatom resource utilization and how cooccurring diatoms adjust their cellular physiology to partition their niche space.American Society for Engineering Education. National Defense Science and Engineering Graduate Fellowshi

Alkaline phosphatase activity in the phytoplankton communities of Monterey Bay and San Francisco Bay

Author Posting. © American Society of Limnology and Oceanography, 2006. This is the author's version of the work. It is posted here by permission of American Society of Limnology and Oceanography for personal use, not for redistribution. The definitive version was published in Limnology and Oceanography 51 (2006): 874–883.Enzyme-labeled fluorescence (ELF) and bulk alkaline phosphatase (AP) activity enzyme assays were used to evaluate the phosphorus (P) status of phytoplankton communities in San Francisco and Monterey bays. Both regions exhibit spatial and temporal variability in bulk AP activity with maximum activities during the early spring and summer periods of high biological productivity. ELF analysis revealed pronounced differences in the makeup of organisms responsible for AP activity in these two environments. In Monterey Bay dinoflagellates are responsible for the bulk of the AP activity. Diatoms infrequently exhibited AP activity. Dinoflagellates that comprised only 14% of all cells counted in Monterey Bay accounted for 78% of AP-producing cells examined. The presence of AP activity in this group suggests that changes in P sources, concentrations, and bioavailability could disproportionably influence this group relative to diatoms in Monterey Bay. In San Francisco Bay, AP production, indicated by ELF, was associated primarily with bacteria attached to suspended particles, potentially used to hydrolyze organic compounds for carbon, rather than to satisfy P requirements. Our results highlight the importance of organic P as a bioavailable nutrient source in marine ecosystems and as a component of the marine P cycle

Microbes and the marine phosphorus cycle

Author Posting. © Oceanography Society, 2007. This article is posted here by permission of Oceanography Society for personal use, not for redistribution. The definitive version was published in Oceanography 20, 2 (2007): 110-116.Phosphorus (P) is fundamental to life, and years of study in marine systems have built a broad understanding of the marine P cycle. Various aspects of marine P biogeochemistry have been reviewed previously (Benitez-Nelson, 2000; Paytan and McLaughlin, 2007). Here, we focus on recent advances in our understanding of marine P and the interactions between microbes and the P cycle. These advances come from a variety of disciplines, but generally highlight three main themes: (1) ocean microbes are adapted for surviving in a variable P environment, (2) the dissolved organic phosphorus (DOP) pool likely plays a critical role in driving growth, metabolism, and community composition of ocean microorganisms, and (3) P is very rapidly cycled, which highlights its importance in marine systems

De novo assembly of Aureococcus anophagefferens transcriptomes reveals diverse responses to the low nutrient and low light conditions present during blooms

Transcriptome profiling was performed on the harmful algal bloom-forming pelagophyte Aureococcus anophagefferens strain CCMP 1850 to assess responses to common stressors for dense phytoplankton blooms: low inorganic nitrogen concentrations, low inorganic phosphorus concentrations, low light levels, and a replete control. The de novo assemblies of pooled reads from all treatments reconstructed ~54,000 transcripts using Trinity, and ~31,000 transcripts using ABySS. Comparison to the strain CCMP 1984 genome showed that the majority of the gene models were present in both de novo assemblies and that roughly 95% of contigs from both assemblies mapped to the genome, with Trinity capturing slightly more genome content. Sequence reads were mapped back to the de novo assemblies as well as the gene models and differential expression was analyzed using a Bayesian approach called Analysis of Sequence Counts (ASC). On average, 93% of significantly upregulated transcripts recovered by genome mapping were present in the significantly upregulated pool from both de novo assembly methods. Transcripts related to the transport and metabolism of nitrogen were upregulated in the low nitrogen treatment, transcripts encoding enzymes that hydrolyze organic phosphorus or relieve arsenic toxicity were upregulated in the low phosphorus treatment, and transcripts for enzymes that catabolize organic compounds, restructure lipid membranes, or are involved in sulfolipid biosynthesis were upregulated in the low light treatment. A comparison of this transcriptome to the nutrient regulated transcriptional response of CCMP 1984 identified conserved responses between these two strains. These analyses reveal the transcriptional underpinnings of physiological shifts that could contribute to the ecological success of this species in situ: organic matter processing, metal detoxification, lipid restructuring, and photosynthetic apparatus turnover

Sinking phytoplankton associated with carbon flux in the Atlantic Ocean

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Limnology and Oceanography 61 (2016): 1172–1187, doi:10.1002/lno.10253.The composition of sinking particles and the mechanisms leading to their transport ultimately control how much carbon is naturally sequestered in the deep ocean by the “biological pump.” While detrital particles often contain much of the sinking carbon, sinking of intact phytoplankton cells can also contribute to carbon export, which represents a direct flux of carbon from the atmosphere to the deep ocean by circumventing the surface ocean food web. Phytoplankton that contributed to carbon flux were identified in sinking material collected by short-term sediment trap deployments conducted along a transect off the eastern shore of South America. Particulate organic carbon flux at 125 m depth did not change significantly along the transect. Instead, changes occurred in the composition and association of phytoplankton with detrital particles. The fluxes of diatoms, coccolithophores, dinoflagellates, and nano-sized cells at 125 m were unrelated to the overlying surface population abundances, indicating that functional-group specific transport mechanisms were variable across locations. The dominant export mechanism of phytoplankton at each station was putatively identified by principal component analysis and fell into one of three categories; (1) transport and sinking of individual, viable diatom cells, (2) transport by aggregates and fecal pellets, or (3) enhanced export of coccolithophores through direct settling and/or aggregationFunding for the DeepDOM cruise was provided by the National Science Foundation (NSF) grant OCE-1154320 to E. B. Kujawinski and K. Longnecker, WHOI. Partial research support was provided by NSF through grants OCE-0925284, and OCE-1316036 to S.T. Dyhrman. C.A. Durkin was supported by a Woods Hole Oceanographic Institution Devonshire Postdoctoral Scholarship

Cross-Basin Comparison of Phosphorus Stress and Nitrogen Fixation in Trichodesmium

We investigated the phosphorus (P) status and N2 fixation rates of Trichodesmium populations from the North Pacific, western South Pacific, and western North Atlantic. Colonies of Trichodesmium were collected and analyzed for endogenous alkaline phosphatase (AP) activity using enzyme-labeled fluorescence ( ELF) and for nitrogenase activity using acetylene reduction. AP hydrolyzes dissolved inorganic phosphate (DIP) from dissolved organic phosphorus and is active in Trichodesmium colonies experiencing P stress. Across multiple stations in the subtropical North and South Pacific, there was low to moderate ELF labeling in Trichodesmium, although labeling was present in other taxa. In contrast, Trichodesmium ELF labeling in the North Atlantic ranged from low to high. Low ELF labeling corresponded with high DIP concentrations while high ELF labeling occurred only at North Atlantic stations with DIP concentrations \u3c = 40 nmol L-1, indicating that Trichodesmium was not experiencing dramatic P stress in the Pacific Ocean while P stress was evident in the western North Atlantic. However, nitrogenase activity was significantly higher in the P-stressed western North Atlantic than in the Pacific Ocean (0.40-1.30 compared to 0.01-0.46 nmol C2H4 h-1 colony-1. These data underscore the differential basin-level importance of P availability to Trichodesmium and suggest that factors other than P are constraining their N2 fixation rates in the Pacific