AbstractSDS-PAGE/silver staining and western blotting were used to investigate the molecular profile of a PBS soluble extract from standard mycelium profile of S. lacrymans FPRL 12C. Both techniques enabled a reference profile for this organism to be established. A polyclonal antiserum raised against standard mycelium of S. lacrymans FPRL 12C was used as the immunological probe.
The profiles of other isolates of S. lacrymans and different Basidiomycetes were compared with FPRL 12C. Protein and antigenic similarity indices enabled differences between these organisms to be quantified. 2 isolates of S. lacrymans differed from FPRL 12 C but to a lesser extent than species in other genera. Lectin staining confrimed these observations and indicated that the majority of antigens were glycoproteins.
SDS-PAGE/silver staining and western blotting allied with similarity indices were found to aid in identification studies which established the identity of one of the anomalous isolates of S. lacrymans. Variations in basic parameters of culture caused minor changes of profile but potentially lethal factors had great effect on profiles. Antigenic differences attributable to growth phase were noted in young and aged mycelium of S. lacrymans.
Protein and antigenic profiles indicated that S. lacrymans in pine and lime sapwood blocks was more similar to young mycelium at low weight loss and to aged mycelium at high weight losses.
Protein and antigenic profiles of field samples of S. lacrymans varied markedly from those of the reference isolate. Protein profiles allied with similarity indices were only appropriate for investigation of a few field mycelial and basidiocarp samples but western blotting identified antigens in most field samples. Identification was made of antigens (a) common to S. lacrymans FPRL 12C and fresh field samples and (b) associated with either fresh or desiccated field samples. The use of these antigens for development of more specific immunological probes is discussed.
|Date of Award||Sep 1992|
|Supervisor||John Palfreyman (Supervisor)|