MICROCHEMICAL TECHNIQUES ON BOTRYTIS CINEREA: CONTRIBUTION TO IDENTIFICATION OF PRIMARY AND SECONDARY METABOLITES

Abstract

Histochemical techniques comprise several staining methods in microscopy for investigation of chemical groups or pathologies regarding tissues in plants and fungi and have been widely used for detection of groups of substances. Thereby, our work demonstrates the application of histochemical techniques recognized for detection of secondary metabolites in plants to detect substances in the fungus Botrytis cinerea. Our findings are compatible with previous reports on metabolites identified in B. cinerea, and show proteins stored in vesicular structures (0.3-0.9µm), accumulated in small groups inside of developed mycelia, and in large quantity in spores (0.2-0.7µm). Alkaloids were only detected in mycelium containing spores, not influenced by their size. Terpenoids were detected in spores, and with lighter color for mycelia vesicles. However, in younger mycelium we verified terpenoids in storage structures in the mycelia segments, demonstrating higher quantity in early development. The presence of neutral polysaccharides and terpenoids was observed around the mycelia containing younger spores and, in some storage, structures close to the spores. Our results corroborate reports on the presence of substances of the secondary metabolism in B. cinerea and could be utilized in other species of fungi to screen substances of pharmacological and nutraceutical interest.