Graduate Thesis Or Dissertation

 

Isolation of protoplasts from selected wood degrading fungi and their uses in studying fungicidal action Public Deposited

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  • Studies of physiological effects of fungicides on fungi are critical for understanding modes of action and mechanism of resistance; however, studies are difficult to perform on filamentous fungi because of the inability to produce homogeneous a biological system using a mass of mycelium. Moreover, the presence of rigid cell wall can limit uptake of chemicals. An alternative approach is to produce protoplasts. A method for isolating protoplasts from common wood-degrading fungi Phanerochatechrysosporium, Postia placenta, Gloeophyllum trabeum, and Trametes versicolor with Novozyme 234 is descibed. Generally, protoplasts were more readily isolated from younger hyphae, while regeneration was better with older hyphae. Fungicidal action was evaluated against Trametes versicolor and Postia placenta using both protoplasts and cell fragments. Generally, similar inhibitory effects were observed with cell viability, glucose utilization, cell respiration, permeability, and copper absorption bioassays. Although protoplasts showed a higher sensitivity to toxicants, this sensitivity was limited compared to that of plant protoplasts. The loss of vulnerable, senescing cells, alteration of some properties of protoplasts during or after lytic enzyme digestion, or biocide absorption by non-viable protoplasts might result in reduced sensitivity of protoplasts to chemicals. The results suggests that further studies are required to delineate more fully the optimum conditions for physiological activity of protoplasts before the full effectivness of the system can be exploited for biocidal action research.
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