Activity of monoterpenoids on the in vitro growth of two Colletotrichum species and the mode of action on C. acutatum

Essential oils and their main compounds, monoterpenoids, are considered as alternative control systems for phytopathogenic fungi, particularly those related to late diseases of fruits and vegetables, like anthracnose caused by Colletotrichum species. In this context, we studied the effect of twenty monoterpenoids on Colletotrichum fructicola and C. acutatum to elucidate their effectiveness and mechanisms of action. Thus, we analyzed mycelial growth and conidial inhibitory concentration, as well as the effect of selected monoterpenoids on membrane integrity and cell vitality, reactive oxygen species (ROS) accumulation, and mitochondrial membrane potential by flow cytometry. The results showed that oxygenated monoterpenoids (alcohols and aldehydes) exhibited higher antifungal activity than their corresponding hydrocarbons, esters, and cyclic counterparts. Indicating that OH^- and O^- radicals react with cellular components affecting fungal homeostasis. In this sense, selected monoterpenoids (citral, citronellol, geraniol, carvacrol, and thymol) inhibited conidial germination of C. acutatum in a dose-dependent manner. The inhibition of conidial germination is associated with a loss of membrane integrity, a decrease of cell metabolism, and a dose-dependent accumulation of ROS, which was non-directly associated with modifications on mitochondrial membrane potential. Membrane dysfunction and ROS accumulation may be responsible for the necrotic behavior induced by high monoterpenoids concentrations, and possible apoptotic response in sub dosages of these compounds.

Keywords: Apoptosis/necrosis; Membrane integrity; ROS; Terpenes.