Protoplast production and transformation of morphological mutants of the Quorn myco-protein fungus, Fusarium graminearum A3/5, using the hygromycin B resistance plasmid pAN7-1
Wiebe. M. G Novakova. M Miller. L Blakebrough. M. L Robson. G. D Punt. P. J Trinci. A. P. J
Mycological Research ; 1997 [Vol.101] Pages:871-877
Abstract
A protocol for the generation of high yields of viable protoplasts has been developed for several highly branched (colonial) strains of the Quorn myco-protein fungus, Fusarium graminearum A3/5. Driselase was found to produce higher protoplast yields (ca 109 g-1 wet weight) than the other lytic enzymes tested (Glucanex, Novozyme, [beta]-glucuronidase, Sigma lytic enzyme, or ICN lytic enzyme), although yields differed for the various strains. Protoplast regeneration frequencies of 25–50% were observed when glucose (1·0 M) or sucrose (1·0 M) was used as the osmotic stabilizer. A highly branched strain of F. graminearum CC1-5, which grows better in submerged culture than the more sparsely branched wild-type strain (A3/5) was transformed using the hygromycin B resistance plasmid pAN7-1.
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