Combination of driselase and lysing enzyme in one molar potassium chloride is effective for the production of protoplasts from germinated conidia of Fusarium verticillioides

• Driselase in 1 M KCl is the first best enzyme for the production of protoplast from F. verticillioides.
• Combined use of driselase and lysing enzyme exhibited the additive effect on protoplast formation
• Germinated conidia of F. verticillioides are the best material for protoplasts production
• Driselase (12.5 mg/ml) and lysing enzyme (10 mg/ml) in 1 M KCl and germinated conidia of F. verticillioides is the best condition for the production of protoplasts
• 1.2 M sucrose is the best osmoticum for the regeneration of protoplasts

Various cell wall degrading enzymes and the protoplasting media were evaluated for the production of protoplast in Fusarium verticillioides. Among the various enzymes tested, driselase at 12.5 mg/ml in 1 M KCl protoplasting medium produced the maximum number of protoplast. Next to driselase, lysing enzyme at 10 mg/ml in 1.2 M MgSO4 protoplasting medium was found to be the second best enzyme for the production of protoplast. More interestingly, the combined use of driselase @ 12.5 mg/ml and lysing enzyme @ 10 mg/ml in 1 M KCl exhibited the additive effect on protoplast formation. Germinated conidia of F. verticillioides are the most susceptible fungal material for protoplast production. The use of sucrose at 1.2 M in the regeneration medium supported the maximum regeneration of protoplast. From the present study, we recommend driselase (12.5 mg/ml) and lysing enzyme (10 mg/ml) in 1 M KCl protoplasting medium and germinated conidia of F. verticillioides for the maximum production of protoplasts and 1.2 M sucrose is the best osmoticum for the regeneration of protoplasts.