ALBERT

Description: The ichthyotoxic and mixotrophic prymnesiophyte Prymnesium parvum is known to produce dense virtually monospecific blooms in marine coastal, brackish, and inshore waters. Fish-killing Pyrmnesium blooms are often associated with macronutrient imbalanced conditions based upon shifts in ambient nitrogen (N):phosphorus (P) ratios. We therefore investigated nutrient-dependent cellular acclimation mechanisms of this microalga based upon construction of a normalized expressed sequence tag (EST) library. We then profiled the transcriptome of P. parvum under nutrient-replete conditions as well as under nitrogen (N) and phosphorus (P) limitation via microarray analyses. Twenty-three genes putatively involved in acclimation to low nutrient levels were identified, among them three phosphate transporters, which were highly upregulated under P-starvation. In contrast, the expression of genes involved in transport and acquisition of ammonium or nitrate/nitrite was unaltered in N-starved cells. We propose that genes upregulated under P- or N-starvation lend themselves as potential tools to monitor nutrient limitation effects at the cellular level and indirectly the potential for initiation and maintenance of toxic blooms of P. parvum.

Description: The toxigenic marine dinoflagellate Alexandrium minutum forms toxic blooms causing paralytic shellfish poisoning (PSP), primarily in coastal waters, throughout the world. We examined effects on physiology and gene expression patterns associated with growth and nutrient starvation in a toxic strain of A. minutum. Bloom-relevant factors, including growth rate, intracellular toxin content, allelochemical activity and nutrient status were investigated in A. minutum cultures grown under different environmental regimes. Allelochemical activity of A. minutum cultures, quantified with a cryptomonad Rhodomonas bioassay, increased with age but was independent of nutrient status. The phenotypic data were integrated and compared with gene expression in cell samples taken at selected points along the growth curve. We observed 489 genes consistently differentially expressed between exponentially growing and growth-limited cultures. The expression pattern of stationary-phase cultures was characterized by conspicuous down-regulation of translation-associated genes, up-regulation of sequences involved in intracellular signalling and some indications of increased activity of selfish genetic elements such as transposons. Treatment-specific patterns included five genes regulated in parallel in all nutrient-limited cultures. The conspicuous decrease in photosynthetic performance identified in N-starved cultures was paralleled by down-regulation of chloroplast-associated genes. The particular gene expression patterns we identified as specifically linked with exponential growth, cessation of growth or nutrient limitation may be suitable biomarkers for indicating the beginning of growth limitation in field- or mesocosm studies.