Effect of cellulase fractionation on the viability of cultured barley (Hordeum vulgare) protoplasts

This is the authors' abstract. We don't add key points for this paper.

The age of the stock plants was important for the barley (Hordeum vulgare L. cv. Perth) protoplast viability. Light conditions under which the stock plants were grown also affected the viability of the protoplasts. Greenhouse‐grown plants yielded much higher number of protoplasts than dark‐grown plants, but protoplast viability was better when protoplasts were isolated from etiolated plants. Light supplied during protoplast culture affected protoplast viability within the first 24 h of culture. Cellulase R‐10 (Onozuka) was better than Cellulysin (Calbiochem) and Cellulase + Macerozyme R‐10 (Onozuka) for barley mesophyll protoplast isolation. Cellulase R‐10 (Onozuka) was fractionated on a G‐75 Sephadex column. The eluted fractions were tested for their ability to release barley mesophyll protoplasts and for their toxicity towards the protoplasts. Only a small part of the Cellulase R‐10 was necessary for protoplast isolation from barley leaves. When the fractionated cellulase was analysed by isoelectric focusing, this part of the cellolase appeared as a single band.