R. Michael L. McKay

Role of iron, light, and silicate in controlling algal biomass in subantarctic waters SE of New Zealand

Phytoplankton processes in subantarctic (SA) waters southeast of New Zealand were studied during austral autumn and spring 1997. Chlorophyll a (0.2–0.3 μg L−1) and primary production (350–650 mg C m−2 d−1) were dominated by cells 1 nmol kg−1, there was little evidence of Fe-stressed algal populations, and Fυ/Fm approached 0.60 at the STC. In addition to these trends, waters of SA origin were occasionally observed within the STC and north of the STC, and thus survey data were interpreted with caution. In vitro Fe enrichment incubations in SA waters resulted in a switch from flavodoxin expression to that of ferredoxin, indicating the alleviation of Fe stress. In another 6-day experiment, iron-mediated increases in chlorophyll a (in particular, increases in large diatoms) were of similar magnitude to those observed in a concurrent Si/Fe enrichment; ambient silicate levels were 4 μM. A concurrent in vitro Fe enrichment, at irradiance levels comparable to the calculated mean levels experienced by cells in situ, resulted in relatively small increases (approximately twofold) in chlorophyll a. Thus, in spring, irradiance and Fe may both control diatom growth. In contrast, during summer, as mean irradiance increases and silicate levels decrease, Fe limitation, Fe/Si colimitation, or silicate limitation may determine diatom growth.

Responses of the photosynthetic apparatus of Dunaliella tertiolecta (Chlorophyceae) to nitrogen and phosphorus limitation

Nitrogen (N) and phosphorus (P) limitation affect the photosynthetic apparatus of Dunaliella tertiolecta in markedly different ways. When grown at 0.25 d−1 (18% of the resource-saturated maximum rate, μmax = 1.39 d−1) in chemostat cultures, N- and P-limited cells were chlorotic relative to nutrient-replete controls. The lutein-to-chlorophyll a ratio increased under both N and P limitation, whereas the neoxanthin-to-chlorophyll a ratio increased only under P limitation. The ratio of accessory photoprotective pigments (α- and β-carotene) to chlorophyll a increased under N-limited conditions. Despite differences in accessory pigment complement, chlorophyll a-specific light absorption coefficients of N- and P-limited cultures did not differ significantly, and were greater than in nutrient-replete conditions. In contrast, the initial slope of the photosynthesis.irradiance (PE) response curve (αChl) declined under nutrient-limiting conditions. There were slight reductions in the maximum quantum efficiency of ph...

Increasing stoichiometric imbalance in North America's largest lake: Nitrification in Lake Superior

more than 600 times the mean requirement ratio for primary producers. We examine the rate of [NO3 ] increase relative to budgets for NO3 and fixed N. Nitrate in Lake Superior has continued to rise since 1980, though possibly at a reduced rate. We constructed whole-lake NO3 and N budgets and found that NO3 must be generated in the lake at significant rates. Stable O isotope results indicate that most NO3 in the lake originated by in-lake oxidation. Nitrate in the lake is responding not just to NO3 loading but also to oxidation of reduced forms of nitrogen delivered to the lake. The increasing [NO 3 ]:[PO4 ]s toichiometric imbalance in this large lake is largely determined by these in-situ processes. Citation: Sterner, R. W., E. Anagnostou, S. Brovold, G. S. Bullerjahn, J. C. Finlay, S. Kumar, R. M. L. McKay, and R. M. Sherrell (2007), Increasing stoichiometric imbalance in North America’s largest lake: Nitrification in Lake Superior, Geophys. Res. Lett., 34, L10406, doi:10.1029/ 2006GL028861.

Lake Superior Supports Novel Clusters of Cyanobacterial Picoplankton

ABSTRACT Very little is known about the biodiversity of freshwater autotrophic picoplankton (APP) in the Laurentian Great Lakes, a system comprising 20% of the worlds lacustrine freshwater. In this study, the genetic diversity of Lake Superior APP was examined by analyzing 16S rRNA gene and cpcBA PCR amplicons from water samples. By neighbor joining, the majority of 16S rRNA gene sequences clustered within the “picocyanobacterial clade” consisting of freshwater and marine Synechococcus and Prochlorococcus. Two new groups of Synechococcus spp., the pelagic Lake Superior clusters I and II, do not group with any of the known freshwater picocyanobacterial clusters and were the most abundant species (50 to 90% of the sequences) in samples collected from offshore Lake Superior stations. Conversely, at station Portage Deep (PD), located in a nearshore urbanized area, only 4% of the sequences belonged to these clusters and the remaining clones reflected the freshwater Synechococcus diversity described previously at sites throughout the world. Supporting the 16S rRNA gene data, the cpcBA library from nearshore station PD revealed a cosmopolitan diversity, whereas the majority of the cpcBA sequences (97.6%) from pelagic station CD1 fell within a unique Lake Superior cluster. Thus far, these picocyanobacteria have not been cultured, although their phylogenetic assignment suggests that they are phycoerythrin (PE) rich, consistent with the observation that PE-rich APP dominate Lake Superior picoplankton. Lastly, flow cytometry revealed that the summertime APP can exceed 105 cells ml−1 and suggests that the APP shifts from a community of PE and phycocyanin-rich picocyanobacteria and picoeukaryotes in winter to a PE-rich community in summer.

Detection and expression of the phosphonate transporter gene phnD in marine and freshwater picocyanobacteria

We describe a PCR-based assay designed to detect expression of the phosphonate assimilation gene phnD from picocyanobacteria. The phnD gene encodes the phosphonate binding protein of the ABC-type phosphonate transporter, present in many of the picocyanobacterial genome sequences. Detection of phnD expression can indicate a capacity of picoplankton to utilize phosphonates, a refractory form of phosphorus that can represent 25% of the high-molecular-weight dissolved organic phosphorus pool in marine systems. Primer sets were designed to specifically amplify phnD sequences from marine and freshwater Synechococcus spp., Prochlorococcus spp. and environmental samples from the ocean and Laurentian Great Lakes. Quantitative RT-PCR from cultured marine Synechococcus sp. strain WH8102 and freshwater Synechococcus sp. ARC-21 demonstrated induction of phnD expression in P-deficient media, suggesting that phn genes are regulated coordinately with genes under phoRB control. Last, RT-PCR of environmental RNA samples from the Sargasso Sea and Pacific Ocean detected phnD expression from the endemic picocyanobacterial population. Synechococcus spp. phnD expression yielded a depth-dependent pattern following gradients of P bioavailability. By contrast, the Prochlorococcus spp. primers revealed that in all samples tested, phnD expression was constitutive. The method described herein will allow future studies aimed at understanding the utilization of naturally occurring phoshonates in the ocean as well as monitoring the acquisition of synthetic phosphonate herbicides (e.g. glyphosate) by picocyanobacteria in freshwaters.

Microbial Distributions and the Impact of Phosphorus on Bacterial Activity in Lake Erie

Abstract In light of recent suggestions concerning the relaxation of controls on phosphorus loading in Lake Erie, and in consideration of our current lack of baseline information on microbial communities in this system, a two-part analysis of the microbial ecology of the lake has been conducted. A comprehensive survey of the surface waters collected data on phytoplankton abundance, bacterial productivity and abundance, and viral abundance that were previously not available for Lake Erie. In parallel, phosphorus amendment experiments were conducted in each of the three hydrologically distinct basins of the lake to determine the effect of increased phosphorus loading on bacterial abundance and productivity. After 72 hours incubation, the addition of phosphorus had a significant impact on the microbial food web. A proliferation of different size classes of phytoplankton was accompanied by increases in bacterial production, but not necessarily bacterial abundance. Observations confirmed previous studies suggesting that the microbial communities in each of the three basins responded differently to the addition of phosphorus. Accompanying nutrient data suggest that the responses were due to limitation by other factors in the presence of excess phosphorus; namely that phytoplankton became nitrogen limited whereas bacteria became carbon limited. These results confirm the importance of the microbial food web in understanding the impact of phosphorus loading on the biotic carbon flow in the Lake Erie ecosystem.

Accumulation of ferredoxin and flavodoxin in a marine diatom in response to Fe

Despite recognition that Fe availability is significant in regulating oceanic production in some regions, the biogeochemistry of this trace element is poorly understood. To complement contemporary methods of analytical chemistry, we have used an immunological approach to monitor the Fe nutrition of marine phytoplankton. In prokaryotes and numerous microalgae, the redox catalyst ferredoxin is functionally replaced by flavodoxin during periods of Fe deficiency. In this study, antibodies were raised against ferredoxin purified from a marine diatom, and their utility as a diagnostic indicator was assessed. A species survey demonstrated broad reactivity with both pennate and centric diatoms and additionally with several nondiatom taxa. In batch cultures of the diatom Phaeodactylum tricornutum Bohlin, in which Fe levels were varied, accumulation of ferredoxin varied with the physiological state of the culture; in unimpaired cells (Fv/Fm≥ 0.65), ferredoxin levels were high, whereas levels dropped markedly in cells experiencing even slight photochemical impairment. Accumulation of flavodoxin varied inversely with that of ferredoxin. An experiment was performed to demonstrate the temporal pattern of accumulation of ferredoxin upon recovery from Fe limitation. Prior to Fe amendment, cells were physiologically impaired (chlorotic, Fv/Fm < 0.3) and contained flavodoxin but no detectable ferredoxin. Following addition of Fe, constraints on photochemistry were relaxed within hours. Coinciding with this, levels of flavodoxin declined, whereas ferredoxin was accumulated to high levels within 8 h.

PHOTOSYNTHESIS AND REGULATION OF RUBISCO ACTIVITY IN NET PHYTOPLANKTON FROM DELAWARE BAY1

Net phytoplankton (> 20 μm) comprised 51 ± 9% of the total chlorophyll (Chl) in a Skeletonema costatum– dominated spring bloom in Delaware Bay. The net phytoplankton had low C:N and high protein: carbohydrate ratios, indicating that their growth was nutrient‐replete. Their photosynthetic responses were characterized by low specific absorption, low light‐limited and light‐saturated rates of photosynthesis, and high quantum yields, indicative of acclimation to low irradiance and internal self‐shading. High fucoxanthin: Chi ratios also indicated low light acclimation, but high photoprotective xanthophyll: Chi ratios suggested a high capacity for photoprotective energy dissipation. Ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) could be activated and deactivated in response to changes in irradiance and was fully activated at the surface of the water column and fully deactivated in aphotic deep water. Maximum Rubisco activity was correlated with Rubisco content and bulk protein content of the phytoplankton and with light‐saturated rates of photosynthesis measured in short (< 20‐min) incubations. Long (60‐min) incubations caused a decrease in the light‐saturated rate of photosynthesis, possibly because of feedback limitation. While feedback limitation is unlikely to occur in the water column, it should be considered when estimating productivity in well‐mixed waters from fixed light‐depth incubations.

Physiological characterization of a Synechococcus sp. (Cyanophyceae) strain PCC 7942 iron-dependent bioreporter for freshwater environments

The complex chemical speciation of Fe in aquatic systems and the uncertainties associated with biological assimilation of Fe species make it difficult to assess the bioavailability of Fe to phytoplankton in relation to total dissolved Fe concentrations in natural waters. We developed a cyanobacterial Fe‐responsive bioreporter constructed in Synechococcus sp. strain PCC 7942 by fusing the Fe‐responsive isiAB promoter to Vibrio harveyi luxAB reporter genes. A comprehensive physiological characterization of the bioreporter has been made in defined Fraquil medium at free ferric ion concentrations ranging from pFe 21.6 to pFe 19.5. Whereas growth and physiological parameters are largely constrained over this range of Fe bioavailability, the bioreporter elicits a luminescent signal that varies in response to Fe deficiency. A dose‐response characterization of bioreporter luminescence made over this range of Fe3+ bioavailability demonstrates a sigmoidal response with a dynamic linear range extending between pFe 21.1 and pFe 20.6. The applicability of using this Fe bioreporter to assess Fe availability in the natural environment has been tested using water samples from Lake Huron (Laurentian Great Lakes). Parallel assessment of dissolved Fe and bioreporter response from these samples reinforces the idea that measures of dissolved Fe should not be considered alone when assessing Fe availability to phytoplankton communities.

VERTICAL MIGRATION BY RHIZOSOLENIA SPP. (BACILLARIOPHYCEAE): IMPLICATIONS FOR FE ACQUISITION

Available data support a mechanism of buoyancy‐mediated vertical migration by large‐sized diatoms of Rhizosolenia spp. as a means to access “new” nitrogen from deep waters. To assess whether phytoplankton simultaneously satisfy their Fe requirements by this mechanism, field samples collected during summer 1996 at stations located along a transect through the central North Pacific gyre were assayed for the presence of flavodoxin and ferredoxin via Western blot analysis. All samples, regardless of their buoyancy status and the station from which they were collected, had accumulated flavodoxin but not ferredoxin. To understand better the significance of the field results, cultures of Rhizosolenia formosa H. Peragallo were grown in the laboratory with varying levels of total Fe (200 nM–10,000 nM). Fe had little effect on the physiological and photochemical parameters measured for each treatment. Growth rates did not exceed 0.17 d−1 and values of Fv/Fm ranged from 0.48 to 0.62. In addition, R. formosa accumulated only flavodoxin at each level of Fe addition. From these results, it appears that for some rhizosolenids, flavodoxin is constitutively expressed. The underlying basis for the constitutive nature of this flavodoxin is unclear at present, although it is likely that it is ultimately related to chronic Fe deficit incurred in natural waters.