Release and regeneration of protoplasts from the fungus Trichothecium roseum

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Abstract:

A protocol for isolating and regenerating protoplasts from Trichothecium roseum has been described. Protoplasts from T. roseum were isolated using (i) a lytic enzyme combination composed of Novozym 234, chitinase, cellulase, and pectinase at a 5-mg/mL concentration and (ii) 0.6 M KCl as an osmotic stabilizer. A maximum number of 28 × 104 protoplasts/mL were obtained at pH 5.5. Experiments on the regeneration and reversion of protoplasts revealed a maximum regeneration (60.8%) in complete medium (potato dextrose – yeast extract agar) amended with 0.6 M KCl. The regenerated protoplasts were similar to the original parent strain in morphology, pigmentation, growth, and sporulation.Key words: Trichothecium roseum, protoplast, isolation, regeneration, lytic enzymes, osmotic stabilizers.

Nous faisons la description d'un protocole d'isolation et de régénération de protoplastes de Trichothecium roseum. Les protoplastes de T. roseum a été isolé avec (i) une combinaison d'enzymes lytiques contenant du Novozym 234, de la chitinase, de la cellulase et de la pectinase à une concentration de 5 mg/mL et (ii) renfermant du chlorure de potassium 0,6 M comme stabilisateur osmotique. Un nombre maximal de 28 × 104 protoplastes/mL a été obtenu à pH 5,5. Des expériences se penchant sur la régénération et la réversion des protoplastes ont signalé une régénération maximale (60,8 %) dans un milieu complet (gélose pomme de terre-dextrose et extrait de levure) enrichi de chlorure de potassium 0,6 M. Les protoplastes régénérés étaient semblables à la souche parentale aux niveaux de la morphologie, de la pigmentation, de la croissance et de la sporulation.Mots clés : Trichothecium roseum, protoplaste, isolation, régénération, enzymes lytiques, stabilizateurs osmotiques.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: March 1, 2003

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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