Abstract:

In this study, the preparation, fusion and regeneration of protoplasts from Agrocybe aegerita and Copyinds comatus were explored. Protoplasts were prepared from the four-day-old mycelia through hydrolysis for 4 h with a mixture of 1% pectase and 1% snailase both in 0.6 mol/L mannitol stabilizing the osmotic pressure (1:1, V/V) at 30 ℃. The optimal protoplast infusion conditions were determined to be: fusion at 30 ℃ for 30 min in the presence of 35% PEG (PEG4000) and 0.2 mol/L CaCl₂ (pH 10) as the fusogenic agents. Inactivated protoplasts were used as the marks to pick fusants out, which were then regenerated on PDA plate medium. After the regeneration of fusants, a comparison of colonial phynotype and antagonistic experiments were made to validate them.

Key words: Agrocybe Aegerita, Copyinds Comatus, protoplast, fusion, regeneration