COLOTELO, N., J. L. SUMNER, and W. S. VOEGELIN. 1971. Chemical studies on the exudate and developing sclerotia of Sclerotirzia sckrotiorlrnl (Lib.) DeBary. Can. J. Microbiol. 17: 1189-1194. The exudate from sclerotia of 6-day-old cultures of Scle~.otinin sclerotiorirnz (Lib.) DeBary was found to contain cations, lipid:, ammonia, amino acids, proteins, and various enzymes. Disc-gel electrophoretic patterns of proteins, peroxidase, and polyphenoloxidase isoenzymes of the exudate differed from those of zclerotial extracts. Specific activities for catalase, peroxidase, polyphenoloxidase, and P-glucosidase enzymes of the exudate and sclerotial extracts were very similar. Cellulolytic and polygalacturonase activities for the exudate and sclerotial extracts were demonstrated. The nature and origin of the exudate are briefly discussed.Numerous small drops of clear liquid are frequently seen on the surface of developing sclerotia in culture, and within a short time these droplets increase greatly in size and may coalesce. Remsberg (1940) noted that the clear liquid from sclerotia of some Typhula species left a crystalline residue when dried on a microslide. This exudate has frequently been referred to by other workers but its chemical composition was not determined. Cooke (1969) fo~ind that the exudate from sclerotia of Sclerotiitia sclerotiorui~l and S. trifolioruin contained soluble carbohydrates, and referred to this exudate as being water that was actively excreted during the rapid development of sclerotia. The exudate from sclerotia of S. sclerotiorum has also been shown to have phenol oxidase activity and contain various salts, amino acids, and proteins (Jones 1970); however, details were not presented. A membrane enveloping the exudate on sclerotia of S. sclerotiorunt was reported by Colotelo et al. (1971) but the origin and nature of this membrane has not yet been determined.In conjunction with our studies on the enzyme changes associated with the development of sclerotia of S. sclerotiorum, the exudate was analyzed for those properties such as pH, dry matter content, inorganic ions, proteins, enzymes, free amino acids, lipids, and free ammonia, which are normally associated with sclerotia and mycelium. Where possible the properties of the exudate and sclerotial extracts were compared.The fungus, S . sclerotior~ml, originally isolated as sclerotia from diseased parsnips, was grown on a medium consisting of 300 g ground carrots, 16 g agar, and 700 ml deionized distilled water. A plug of inoculum, 7 mnl in diameter, from the periphery of a young colony grown o n the above medium was placed in the center of each Petri plate 138 mnl in diameter containing about 60ml of the carrot-agar medium. Cultures were incubated at 20°C.The liquid o n the surface of sclerotia of 6-day-old cultures was collected with a Pasteur pipette and analyzed immediately after being passed through a sterile 0.45-br Millipore filter. Sclerotia from these cultures were extratced using the following procedure and used for the electrophoretic stud...
Liquid droplets commonly observed on developing sclerotia of Sclerotinia sclerotiorum were found to be enveloped by small sacs. These sacs were present at all stages of sclerotia development when the fungus was cultured on various media and at various temperatures.
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