Summary
The conditions for protoplast isolation and reversion inPleuro tus sajor-caju were determined. Spherical, osmotically sensitive protoplasts were released from the mycelia through the action of Novozym 234 and cellulase CP in a suitable osmotic solution. A maximum protoplast yield was obtained from young mycelia at an early exponential growth phase by the use of 0.6 mol/l MgSC4 in a 0.01 mol/l phosphate buffer at pH 5.0. Mannitol appeared to be more suitable forPleurotus protoplast reversion. Two patterns of morphological development of the protoplasts were observed in the osmotic liquid medium. The reversion frequency was, however, low at 4–5%. There appeared to be no significant difference in the reversion frequencies of the protoplasts grown on the complete medium and on the minimal medium.
Résumé
Isolement et réversion des protoplastes de Pleurotus sajor-caju Les conditions pour l'isolement de protoplastes dePleurotus sajor-caju et pour leur réversion ont été déterminées. Des protoplastes sphériques et osmotiquement sensibles ont été préparés à partir du mycélium par action de Novozym 234 et de cellulase CP en milieu osmotiquement approprié. Le rendement maximum en protoplastes est obtenu à partir du mycélium jeune, en phase exponentielle précoce, dans un tampon phosphates 0,01 M à pH 5,0, contenant MgSO4 0,6 M. Pour la réversion des protoplastes, l'agent le plus favorable est le manitol. Deux modes de développement morphologique des protoplastes en milieu osmotique ont été constatés. Toutefois, la fréquence de réversion est faible (4 à 5%). Il ne semble pas que la fréquence de réversion soit significativement différente lorsque les protoplastes sont cultivés en milieu complet ou en milieu minimal.
Resumen
Aislamiento y reversión de protoplastos en Pleurotus sajor-caju Se determinaron las condiciones para el aislamiento y reversión de protoplastos enPleurotus sajor-caju. Protoplastos esféricos y osmóticamente sensibles se obtuvieron a partir de micelio mediante la action de Novozym 234 y celulasa CP en un medio osmótico adecuado. El máximo numéro de protoplastos fueron obtenidos a partir de miclio joven en la primera parte de la fase exponencial de crecimiento utilizando 0.6 mol/l MgSO en un tampón fosfato (0.01 mol/l) a pH 5.0. Manitol parece ser adecuado para la reversion dePleurotus. Se observaron dos patrones de desarrollo morfológico de los protoplastos en el medio osmótico líquido. La frecuencia de reversion fue, sin embargo, baja (4–5%). No se observaron diferencias significativas entre los protoplastos obtenidos en el medio completo y los que lo fueron en un medio mínimo.
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This work is part of the first author's PhD thesis submitted to the Department of Biology of the Chinese University of Hong Kong.
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Lau, W.C., Dhillon, E.K.S. & Chang, S.T. Isolation and reversion of protoplasts ofPleurotus sajor-caju . World J Microbiol Biotechnol 1, 345–353 (1985). https://doi.org/10.1007/BF01553419
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DOI: https://doi.org/10.1007/BF01553419