Factors affecting the production and regeneration of protoplasts from Colletotrichum lindemuthianum

Ciência e Agrotecnologia

Francine Hiromi Ishikawa²  Quélen De Lima Barcelos¹  Elaine Aparecida De Souza²  Eustáquio Souza Dias² 

[1] ,Universidade Federal de Lavras Departamento de Biologia Lavras MG

关键词: Protoplast formation;    lytic enzyme;    anthracnose;    osmotic stabilizer;    filamentous fungi;    Formação de protoplastos;    enzima lítica;    antracnose;    estabilizador osmótico;    fungo filamentoso;   

DOI  :  10.1590/S1413-70542010000100009

来源: SciELO

【 摘 要 】

The present work reports factors affecting the production and regeneration of protoplasts from Colletotrichum lindemuthianum. The usefulness of protoplast isolation is relevant for many different applications and has been principally used in procedures involving genetic manipulation. Osmotic stabilizers, lytic enzymes, incubation time and mycelial age were evaluated in terms of their effects on protoplast yield. The optimal condition for protoplast production included the incubation of young mycelia (48 h) in 0.6 mol l-1 NaCl as the osmotic stabilizer, with 30 mg ml-1 Lysing Enzymes from Trichoderma harzianum for 3 h of incubation. In these conditions protoplasts production was higher than 10(6) protoplatos ml-1 in the digestion mixture, number suitable enough for experiments of transformation in fungi. Sucrose concentrations of 1.2 mol l-1 and 1 mol l-1 were the most suitable osmotic stabilizers for the regeneration after 48 h, with rates of 16.35% and 14.54%, respectively. This study produced an efficient method for protoplast production and reverted them into a typical mycelial morphology using a Colletotrichum lindemuthianum LV115 isolate.

【 授权许可】

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