Use of enzymes results in a high yield of uniform protoplasts after removal of cellular debris

Protoplasts can originate from different sources: greenhouse or field material, micropropagated plants, calli, ...

Freshly isolated protoplasts

• obtain sterile plant material
• rinsing in a suitable osmoticum
• facilitating enzyme penetration
• sequential or mixed-enzyme treatment
• purification of the isolated protoplasts (removal of enzymes and cellular debris)

transfer to a suitable medium

>A typical procedure
to obtain mesophyll
protoplasts