RESUMO
BACKGROUND: In initial assays, Thymus vulgaris essential oil (TEO) has demonstrated activity against several plant-pathogenic fungi and has reduced the fungal diseases to levels comparable with commercial fungicides. Thus, the goal of this work was to identify the mode of action in fungi of TEO and its major compound thymol (TOH) at the cellular level using an ultrastructure approach. RESULTS: TEO from leaves and TOH had minimum inhibitory concentrations (MICs) of 500 and 250 µg mL(-1) respectively against A. alternata; under the same conditions, MIC for a commercial fungicide was 1250 µg mL(-1) . Ultrastructure analysis showed that TOH phenolic substance prevented fungal growth, reduced fungal viability and prevented the penetration in fruits by a cell wall/plasma membrane interference mode of action with organelles targeted for destruction in the cytoplasm. Such mode of action differs from protective and preventive-curative commercial fungicides used as pattern control. CONCLUSION: These findings suggest that TOH was responsible for the antifungal activity of TEO. Therefore, both the essential oil and its major substance have potential for use in the development of new phenolic structures and analogues to control Alternaria brown spot disease caused by Alternaria alternata.
Assuntos
Alternaria/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Viabilidade Microbiana/efeitos dos fármacos , Óleos Voláteis/farmacologia , Thymus (Planta)/química , Alternaria/crescimento & desenvolvimento , Alternaria/fisiologia , Citrus/microbiologia , Testes de Sensibilidade Microbiana , Doenças das Plantas/microbiologia , Timol/farmacologiaRESUMO
BACKGROUND: In previous studies, the extract from Anadenanthera colubrina was active against Alternaria alternata in vitro and reduced the disease caused by this fungus on Murcott tangor fruits to levels that have been obtained using commercial fungicides. Therefore, the goal of the present work was to isolate and identify the active substances in this extract and identify in silico their protein target in the fungus. RESULTS: The bioguided fractionation of the methanol extract from the fruits of A. colubrina resulted in the isolation of ß-sitosterol and ß-sitosteryl linoleate, which had minimal inhibitory concentrations (MICs) of 250 and 500 µg mL(-1) , respectively, against A. alternata. Under the same conditions, the MICs for two commercial fungicides were 1250 and 19 µg mL(-1) . In silico studies showed that these steroidal substances bind well to oxysterol-binding proteins from Saccharomyces cerevisiae. CONCLUSION: ß-Sitosterol and ß-sitosteryl linoleate, produced by A. colubrina, are active against A. alternata. In silico studies suggest that these substances may act by binding to oxysterol-binding proteins. Therefore, both substances and these proteins have potential use in the development of new steroidal structures and analogues to control the disease caused by A. alternata.