Simplified growth media for Entomophthora egressa protoplasts

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Abstract
The protoplast stage of Entomophthora egressa utilized cysteic acid, L-isoleucine, L-leucine, L-methionine, L-proline, DL-serine, L-threonine, L-glutamine and (or) L-asparagine, and L-valine during the initial 16 h of incubation in modified Grace's medium but did not utilize glucose. Medium A, containing the above amino acids (excluding L-isoleucine) plus L-tyrosine, L-glutamic acid, L-aspartic acid, and L-lysine, supported normal growth and cell wall regeneration. The amino acid composition was further simplified to include only L-glutamine, L-asparagine, and L-methionine (medium B). The growth rates and morphogenesis of the protoplasts in media A and B were compared with those in medium A plus albumin. Fetal calf serum was essential for protoplast growth in shaken cultures in both simplified media; however, it was not required for growth in stationary cultures. Depending on the medium employed, E. egressa exhibited complete or partial vitamin auxoautotrophy.