Different stages of the automortality in phytoplankton have been studied applying flow cytometry. These stages are, in order of expression: (1) compromised cell membranes, (2) degradation of the photosynthetic pigments and reduction of the photosynthetic activity, (3) fragmentation of the genomic DNA. The integrity test of the cell membranes is based on the inability of the DNA-specific stain SYTOX Green to pass into cells with intact plasma membranes. The reduction in photosynthetic activity was examined by sorting 14C-labelled phytoplankton cells differing in viability. Finally, DNA fragmentation was traced by measuring changes in genomic DNA. The different phytoplankton species tested showed a great variety in response when grown under the same conditions, but there was also considerable intraspecific variation. Unstained cells, fully stained cells (equivalent to full staining of genomic DNA in fixed cells) and cells with intermediate fluorescence signal occurred together within the same culture. The photosynthetic activity in cells with a reduced viability dropped by as much as 60% relative to that of the viable cells. In the subsequent stage, when photosynthetic pigments were fully degraded, this value dropped further to around 10%. Cells in this stage also showed subdiploidy as a result of genome fragmentation. Field tests using samples of phytoplankton collected in the North Atlantic Ocean (40° N, 23° W) during spring showed staining properties similar to those found in cultures grown at suboptimal growth conditions. The percentage of non-viable cells varied considerably (ranging from 5% to 60%) between the various phytoplankton groups present. The lowest value was observed for Synechococcus, but some pico-eukaryotes showed percentages as high as 60%. Moreover, the viability varied with depth (light level) and over a light–dark cycle. The present findings suggest the existence of a (genetically based) uniform process of automortality in phytoplankton. Non-viable cells are a substantial component of the oceanic phytoplankton, affecting the food-web structure and species succession.

The past decade has seen substantial breakthroughs in understanding the biochemistry, molecular biology and regulation of nitrate reductases (NR) in higher plants and green algae. In contrast, although there has been considerable interest in using various measurements of NR to provide ecophysiological information, comparable knowledge of NR is largely lacking in algal groups other than chlorophytes. Applying information about NR from chlorophytes and higher plants to other algae may be difficult. There is evidence that non-chlorophyte forms of NR are diverse and distinct in terms of biochemical characteristics and regulatory features. Key areas to be pursued in non-chlorophyte algae include the identification and adoption of model organisms for NR research in different algal groups; the creation of selected and engineered mutants; the purification, biochemical characterization and production of antibodies to different algal forms of NR; the identification of NR genes; and the undertaking of coordinated research on nitrate uptake proteins and other enzymes in pathways of nitrate assimilation.