Isolation and regeneration of Penicillium brevicompactum protoplasts

ABSTRACT. The isolation and regeneration of fungal protoplasts is a key step for the establishment of transformation systems, electrophoretic karyotype analysis and fusion between strains, all techniques of broad application on improvement programs of filamentous fungi. To establish conditions for the isolation and regeneration of Penicillium brevicompactum protoplasts, that is an excellent pectinase producer, different lytic enzymes and osmotic stabilizers were tested. P. brevicompactum protoplasts were obtained at a larger scale when their mycelium was digested with 15mg.mL-1 of Glucanex (Novo Nordisk) and 0.8 mol.L–1 NaCl as osmotic stabilizer. The best osmotic stabilizer for the regeneration of P. brevicompactum protoplasts was 0.8 mol.L–1 KCl, with a regeneration frequency of 36.58%. This protocol can be applied in genetic analysis of this Penicillium species which, to date has been poorly characterized,. Key words: Penicillium brevicompactum, pectinases; protoplasts. RESUMO. Isolamento e regeneração de protoplastos de Penicillium brevicompactum. O isolamento e regeneração de protoplastos de fungos é um passo fundamental para o estabelecimento de sistemas de transformação, análise do cariótip