Cells of the green microalga Haematococcus pluvialis were induced to accumulate the ketocarotenoid pigment, astaxanthin. This induction was achieved by the application of the following environmental conditions: light intensity (170 μmol m~⁻²s⁻¹), phosphate starvation and salt stress (NaCl 0.8%). These conditions retarded cell growth as reflected by a decrease in cell division rate, but led to an increase in astaxanthin content per cell. Accumulation of astaxanthin required nitrogen and was associated with a change in the cell stage from biflagellate vegetative green cells to non-motile and large resting cells. It is suggested that environmental or nutritional stresses, which interfere with cell division, trigger the accumulation of astaxanthin. Indeed, when a specific inhibitor of cell division was applied, a massive accumulation of astaxanthin occurred.

Wild-type cyanobacteria of the genus Anabaena are capable of oxygenic photosynthesis, differentiation of cells called heterocysts at semiregular intervals along the cyanobacterial filaments, and aerobic nitrogen fixation by the heterocysts. To foster analysis of the physiological processes characteristic of these cyanobacteria, we have constructed a family of shuttle vectors capable of replication and selection in Escherichia coli and, in unaltered form, in several strains of Anabaena. Highly efficient conjugative transfer of these vectors from E. coli to Anabaena is dependent upon the presence of broad host-range plasmid RP-4 and of helper plasmids. The shuttle vectors contain portions of plasmid pBR322 required for replication and mobilization, with sites for Anabaena restriction enzymes deleted; cyanobacterial replicon pDU1, which lacks such sites; and determinants for resistance to chloramphenicol, streptomycin, neomycin, and erythromycin.

Scaling up algal cultures to the very large volumes required for commercial production is a complex task and requires skilled and experienced personnel. First it is necessary to consider how to optimize the process of producing enough inoculum for the large ponds or photobioreactors in order to minimize the time and cost required. In order to minimize the need for re-inoculation from stock cultures it is also essential to manage the large-scale cultures to avoid significant contamination or collapse. The maintenance of long-term, stable, high-productivity, large-scale cultures, usually under prevailing outdoor conditions of variable irradiance, temperature and rainfall, presents additional challenges most of which are not seen in the constant environment experienced by small-scale laboratory cultures. Methods and protocols to deal with these can only be developed at the large-scale and they will mostly be specific for the alga being cultured, the culture system being used and the location of the production plant. A common feature of all large-scale operations known to us is that, over time (years), both productivity and reliability of the cultures improve as the operators gather experience in managing their cultures.