Release of single cells from the colonial oil-producing alga Botryococcus braunii by chemical treatments.

We tested for chemical reagents that would be useful in preparing a large number of vital single cells from colonial Botryococcus braunii B-race, variety Showa. Among the 18 reagents assayed, glycerol and erythritol showed the highest potency for releasing single cells. Incubation in medium containing these reagents released 40-50 % single cells in 15 min. Fluorescent staining with Nile red revealed that except for the cap-like structures the released single cells were free of hydrocarbon oils that accumulated in the extracellular matrix where the single cells were embedded. However, to maintain the prepared single cells in vital condition, they must be maintained at a high concentration (>2 × 10(7) cells/ml); at low concentrations, they rapidly lost chlorophyll and get disrupted. In contrast to the above results obtained using B-race, Showa, single cells prepared from A-race varieties survived even at low cell concentrations.