Project description:A functional gene microarray was developed and used to investigate phytoplankton community composition and gene expression in the English Channel. Genes encoding the CO2 fixation enzyme RuBisCO (rbcL) and the nitrate assimilation enzyme nitrate reductase (NR) representing several major groups of phytoplankton were included as oligonucleotide probes on the 'phytoarray'. Five major groups of eukaryotic phytoplankton that possess the Type 1D rbcL gene were detected, both in terms of presence (DNA) and activity (rbcL gene expression). Changes in relative signal intensity among the Type 1D rbcL probes indicated a shift from diatom dominance in the spring bloom to dominance by haptophytes and flagellates later in the summer. Because of the limitations of a smaller database, NR probes detected fewer groups, but due to the greater diversity among known NR sequences, NR probes provided higher phylogenetic resolution than did rbcL probes, and identified two uncultivated diatom phylotypes as the most abundant (DNA) and active (NR gene expression) in field samples. Unidentified chlorophytes and the diatom Phaeodactylum tricornutum were detected at both the DNA and cDNA (gene expression) levels. The reproducibility of the array was evaluated in several ways and future directions for further improvement of probe development and sensitivity are outlined. The phytoarray provides a relatively high resolution, high throughput approach to assessing phytoplankton community composition in marine environments. Keywords: seawater natural assemblages, functional gene expression Two functional genes, nitrate reductase and RuBisCO, 4 - 8 replicate features per array
Project description:A functional gene microarray was developed and used to investigate phytoplankton community composition and gene expression in the English Channel. Genes encoding the CO2 fixation enzyme RuBisCO (rbcL) and the nitrate assimilation enzyme nitrate reductase (NR) representing several major groups of phytoplankton were included as oligonucleotide probes on the 'phytoarray'. Five major groups of eukaryotic phytoplankton that possess the Type 1D rbcL gene were detected, both in terms of presence (DNA) and activity (rbcL gene expression). Changes in relative signal intensity among the Type 1D rbcL probes indicated a shift from diatom dominance in the spring bloom to dominance by haptophytes and flagellates later in the summer. Because of the limitations of a smaller database, NR probes detected fewer groups, but due to the greater diversity among known NR sequences, NR probes provided higher phylogenetic resolution than did rbcL probes, and identified two uncultivated diatom phylotypes as the most abundant (DNA) and active (NR gene expression) in field samples. Unidentified chlorophytes and the diatom Phaeodactylum tricornutum were detected at both the DNA and cDNA (gene expression) levels. The reproducibility of the array was evaluated in several ways and future directions for further improvement of probe development and sensitivity are outlined. The phytoarray provides a relatively high resolution, high throughput approach to assessing phytoplankton community composition in marine environments. Keywords: seawater natural assemblages, functional gene expression
Project description:Atrazine is one of the most commonly used herbicide and has been frequently detected in estuarine and offshore waters worldwide. As a photosystem Ⅱ inhibitor, atrazine may inhibit phytoplankton from fixating of CO2 and alter its carbon metabolism, which will undoubtedly have negative effect on the primary productivity and carbon sequestration capacity of coastal waters. However, the existing reports mainly focused on agriculture and freshwater ecosystems and are mostly toxicity test with high-dose of atrazine, which have little concern about the negative effects of atrazine on the carbon metabolism of phytoplankton and can’t reflect the actual toxic situation in offshore water. Diatoms are widely distributed in freshwater and oceans and contribute at least 20% of the global CO2 assimilation, which is an ideal model group to assess the ecological risk of atrazine. Here we present a comprehensive analysis of the physiological and genome-wide gene expression characteristics of the diatom P. tricornutum Pt-1 (CCMP 2561) treated with environmental dose of atrazine at different stress stages.
Project description:As an essential micronutrient that is scarce in surface ocean waters, zinc (Zn) has the potential to limit oceanic photosynthetic productivity and influence the global carbon cycle. Here we observed Zn co-limitation with iron (Fe) in the natural phytoplankton community of Terra Nova Bay, Antarctica, induced by the drawdown of seawater CO2 and dZn during a bloom. Incubations amended with Zn resulted in significantly higher chlorophyll a content and greater macronutrient and dissolved inorganic carbon drawdown compared to Fe addition alone. Multiple Zn and Fe response proteins were observed in experimental and water column samples demonstrating co-stress in various algal taxa. Together these results demonstrate that Zn limitation can occur in productive Antarctic coastal ecosystems. Thus, Zn may be an important factor limiting the total productivity potential of marine phytoplankton.