Summary
Protoplast yields from Sclerotium glucanicum using Novozym 234 as the lytic enzyme were affected by the osmotic stabilizers selected, the incubation conditions used for wall degradation, and culture age. Scanning electron microscopic observations revealed that protoplast release from all hyphal regions gradually followed random wall attack, and nuclear staining showed that some protoplasts contained as many as eight nuclei. Their regeneration involved germ tube production on solid media, but formation of chains of buds and possibly cytoplasmic cleavage in liquid medium. Regenerated protoplasts gave similar exopolysaccharide yields to those of the parent culture.
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References
Anne J, Eyssen H, De Somer P (1974) Formation and regeneration of Penicillium chrysogenum protoplasts. Arch Microbiol 98:159–166
Brown JA, Ogawa K, Wood TM (1986) Studies on the preparation and regeneration of protoplasts from the cellulolytic fungus Penicillium pinaphilum. Enzyme Microb Technol 9:527–532
Browne HM, Dowsett AB, Gibson RK, Peberdy JF (1987) The isolation and regeneration of protoplast from Lipomyces starkeyi: an oleaginous yeast. J Microbiol Methods 6:153–165
Davis B (1986) Factors influencing protoplast isolation. In: Peberdy JF, Ferenzy L (eds) Fungal protoplasts: applications in biochemistry and genetics. Dekker, New York, pp 45–71
Gull K, Trinci APG (1974) Detection of areas of wall differentiation in fungi using fluorescent staining. Arch Microbiol 96:53–57
Hamlyn PF, Bradshaw RE, Mellon FM, Santiago CM, Wilson JM, Peberdy JF (1981) Efficient protoplast isolation from fungi using commercial enzymes. Enzyme Microb Technol 3:321–325
Isaac S (1985) Metabolic properties of protoplasts. In: Peberdy JF, Ferenzy L (eds). Fungal protoplasts: applications in biochemistry and genetics. Dekker, New York, pp 171–187
Johnson J, Kirkwood S, Misaki A, Nelson TE, Scarletti JV, Smith F (1963) Structure of a new glucan. Chem Ind (Lond) 41:820–822
Kropp BR, Fortin JA (1986) Formation and regeneration of protoplasts from the ectomycorrhizal basidiomycete Laccaria bicolor. Can J Bot 64:1224–1226
Lynch PT, Collin HA, Isaac S (1985) Isolation and regeneration of protoplasts from Fusarium tricinctum and F. oxysporum. Trans Brit Mycol Sco 85:135–140
Michel M, Seviour RL, Pethica LM (1987) Exocellular polysaccharide production by isolates of Epicoccum purpurascens. Biotechnol Lett 9:741–744
Peberdy JF (1979) Fungal protoplasts: isolation reversion and fusion. Ann Rev Microbiol 33:21–39
Picataggio SK, Schamhart DHJ, Montenecourt BS, Eveleigh DE (1983) Sphaeroplast formation and regeneration in Trichoderma reesei. Eur J Appl Microbiol Biotechnol 17:121–128
Quigley DR, Taft CS, Stark T, Selitrennikoff CP (1987) Optimal conditions for the release of protoplasts of Neurospora using Novozym 234. Exp Mycol 11:236–240
Rodgers NE (1973) Scleroglucan. In: Whistler RL (ed) Industrial gums. Academic Press, New York, pp 499–511
Seviour RJ, Pethica LM, McClure S (1984) A simple modified procedure for preparing microbial cells for scanning electron microscopy. J Microbiol Methods 3:1–5
Thomas KR, Davis B (1980) The effect of calcium on protoplast release from species of Aspergillus. Microbios 28:69–80
Thomas KR, Davis B, Mills J (1984) Ultrastructural observations of protoplast release from Aspergillus fumigatus. Trans Brit Mycol Soc 82:405–411
Wollingen P van, Seviour RJ (1986) Using the stomacher for preparing standard inocula from non-sporeforming fungi. Trans Brit Mycol Soc 86:487–490
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Deed, A., Seviour, R. Formation and regeneration of protoplasts of Sclerotium glucanicum . Appl Microbiol Biotechnol 31, 259–264 (1989). https://doi.org/10.1007/BF00258406
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DOI: https://doi.org/10.1007/BF00258406