Protoplasts from zoospores and cysts of Haematococcus pluvialis alga (Chlorophyta, Volvocales)

Two methods for preparing protoplasts from Haematococcus pluvialis Flotow are reported. This microalga is characterized by two types of cells: biflagellated vegetative zoospores and cysts. For zoospores, we used a proteinase K (0.06%). The cellulose nature of cyst walls was demonstrated by staining with calcofluor-white fluorochrome. A mixture of cellulase (1%) and pectolyase (0.5%) was effective on cysts. Cell wall degradation during enzymatic incubation was studied by optical, electron, and fluorescence microscopy. Cellulase and pectolyase were more specific and efficient than proteinase K.