Factors That Influence the Yield and Viability of Cucumber (Cucumis sativus L) Cotyledon Protoplasts

Abstract

Protoplasts isolated from cotyledons of aseptically germinated cucumber seedlings were divided into three size classes. The relationships between tissue age, isolation procedure, yield and protoplast size were investigated. During germination and up to an age of 13 days, the percentage of protoplasts in each size class underwent considerable change with a big reduction in percentage of the largest protoplasts in older cotyledons. Protoplast size and yield could also be manipulated by varying the isolation technique. In this context, temperature, incubation time and shaker speed were significant. By selecting tissue of appropriate age and using a carefully selected isolation procedure the percentage of viable cucumber protoplasts with the ability to form a cell wall and divide can be increased considerably.