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In an effort to better understand the diversity of genes coding for nitrogen (N) uptake and assimilation pathways among microalgae, we analyzed the transcriptomes of five phylogenetically diverse single celled algae originally isolated from the same high arctic marine region. The five photosynthetic flagellates (a pelagophyte, dictyochophyte, chrysoph-yte, cryptophyte and haptophyte) were grown on standard media and media with only urea or nitrate as a nitrogen source; cells were harvested during late exponential growth. Based on homolog protein sequences, transcriptomes of each alga were interrogated to retrieve genes potentially associated with nitrogen uptake and utilization pathways. We further investigated the phylogeny of poorly characterized genes and gene families that were identified. While the phylogeny of the active urea transporter ( DUR3) was taxonomically coherent, those for the urea transporter superfamily, putative nitrilases and amidases indicated complex evolutionary histories, and preliminary evidence for horizontal gene transfers. All five algae expressed genes for ammonium assimilation and all but the chrysophyte expressed genes involved in nitrate utilization and the urea cycle. Among the four algae with nitrate transporter transcripts, we detected lower expression levels in three of these (the dictyochophyte, pelagophyte, and cryptophyte) grown in the urea only medium compared with cultures from the nitrate only media. The diversity of N pathway genes in the five algae, and their ability to grow using urea as a nitrogen source, suggest that these flagellates are able to use a variety of organic nitrogen sources, which would be an advantage in an inorganic nitrogen - limited environment, such as the Arctic Ocean. [ABSTRACT FROM AUTHOR]

Metabolic diversity of heterotrophic bacterioplankton was tracked from early winter through spring with Biolog Ecoplates under the seasonally ice covered arctic shelf in the Canadian Arctic (Franklin Bay, Beaufort Sea). Samples were taken every 6 days from December 2003 to May 2004 at the surface, the halocline where a temperature inversion occurs, and at 200 m, close to the bottom. Despite the low nutrient levels and low chlorophyll a, suggesting oligotrophy in the winter surface waters, the number of substrates used (NSU) was greater than in spring, when chlorophyll a concentrations increased. Denaturing gradient gel electrophorisis analysis also indicated that the winter and spring bacterial communities were phylogenetically distinct, with several new bands appearing in spring. In spring, the bacterial community would have access to the freshly produced organic carbon from the early phytoplankton bloom and the growth of rapidly growing specialist phenotypes would be favoured. In contrast, in winter bacterioplankton consumed more complex organic matter originated during the previous year's phytoplankton production. At the other depths we tested the NSU was similar to that for the winter surface, with no seasonal pattern. Instead, bacterioplankton metabolism seemed to be influenced by resuspension, advection, and sedimentation events that contributed organic matter that enhanced bacterial metabolism. [ABSTRACT FROM AUTHOR]

Our pigment analyses from a year-long study in the coastal Beaufort Sea in the western Canadian Arctic showed the continuous prevalence of eukaryotic picoplankton in the green algal class Prasinophyceae. Microscopic analyses revealed that the most abundant photosynthetic cell types were Micromonas-like picoprasinophytes that persisted throughout winter darkness and then maintained steady exponential growth from late winter to early summer. A Micromonas (CCMP2099) isolated from an Arctic polynya (North Water Polynya between Ellesmere Island and Greenland), an ice-free section, grew optimally at 6°C–8°C, with light saturation at or below 10 μmol photons·m−2·s−1 at 0°C. The 18S rDNA analyses of this isolate and environmental DNA clone libraries from diverse sites across the Arctic Basin indicate that this single psychrophilic Micromonas ecotype has a pan-Arctic distribution. The 18S rDNA from two other picoprasinophyte genera was also found in our pan-Arctic clone libraries: Bathycoccus and Mantoniella. The Arctic Micromonas differed from genotypes elsewhere in the World Ocean, implying that the Arctic Basin is a marine microbial province containing endemic species, consistent with the biogeography of its macroorganisms. The prevalence of obligate low-temperature, shade-adapted species in the phytoplankton indicates that the lower food web of the Arctic Ocean is vulnerable to ongoing climate change in the region. [ABSTRACT FROM AUTHOR]

Heterotrophic flagellates play fundamental roles in marine ecosystems as picoplankton grazers. This recognized importance contrasts with our ignorance of the taxonomic composition of this functional group, which remains mostly unidentified by microscopical and culturing approaches. Recent molecular marine surveys based on 18S rDNA genes have retrieved many sequences unrelated to cultured organisms and marine stramenopiles were among the first reported uncultured eukaryotes. However, little is known about the organisms corresponding to these sequences. Here we determine the abundance of several marine stramenopile lineages in surface marine waters using molecular probes and fluorescent in situ hybridization. We show that these protists are free-living bacterivorous heterotrophic flagellates. They were widely distributed, occurring in the five world oceans, and accounted for a significant fraction (up to 35%) of heterotrophic flagellates in diverse geographic regions. A single group, MAST-4, represented 9% of cells within this functional assemblage, with the intriguing exception of polar waters where it was absent. MAST-4 cells likely contribute substantially to picoplankton grazing and nutrient re-mineralization in vast areas of the oceans and represent a key eukaryotic group in marine food webs. [ABSTRACT FROM AUTHOR]

Bacterivory was determined in surface waters of Franklin Bay, western Arctic, over a seasonal ice-covered period (winter-spring, 2003-2004). The objectives were to obtain information on the functioning of the microbial food web under the ice, during winter (from 21 December 2003 to 21 March 2004) and during spring (from 22 March 2004 to 29 May 2004), and to test whether bacterial losses would increase after the increase in bacterial production following the spring phytoplankton bloom. Chl a concentrations ranged from 0.04 to 0.36 µg L-1, increasing in March and reaching a peak in April. Bacterial biomass showed no consistent trend for the whole period, and protist biomass followed a pattern similar to that of Chl a. Bacterial production increased 1 week after Chl a concentrations started to increase, while bacterivory rates increased very slightly. Average bacterivory rates in winter (0.16 ± 0.07 µg C L-1 d-1) were not significantly different from those in spring (0.29 ± 0.24 µg C L-1 d-1). Average bacterial production, on the other hand, was similar to bacterivory rates in winter (0.19 ± 0.38 µg C L-1 d-1), but higher than bacterivory in spring (0.93 ± 0.28 µg C L-1 d-1). Therefore, bacterial production was controlled by grazers during winter and by substrate concentration in spring. [ABSTRACT FROM AUTHOR]