Inhibitory activity of 2-deoxy-D-glucose and Candida saitoana against Penicillium digitatum

The toxic activity of 2-deoxy-D-glucose (2-DG) alone or combined with the biocontrol yeast Candida saitoana strain 8C was evaluated in vitro and in vivo against the postharvest fungal pathogen Penicillium digitatum. In order to assess the effect of the 2-DG on both the biocontrol yeast and fungal pathogen, in vitro tests were performed in Petri dishes containing potato dextrose agar amended with different concentrations (1.5, 3.0, 6.0, 15.0, 30.0, 60.0 mM) of the sugar. The plates were then seeded with 25 microl of a P. digitatum conidial suspension at 10(5) conidia/mL. Result of the assays showed an enhanced inhibitory activity as concentration increased from 15.0 to 60.0 mM. Corroborated by SEM observations showing a reduced growth and the appearance of damaged hyphae were found. At 60 mM of 2-DG, a total inhibition occurred while concentrations from 1.5 to 6.0 mM resulted ineffective. The same tests evidenced no adverse effects on the yeast 8C at all tested concentrations. In vivo assays were carried out on orange fruit cv 'Biondo comune', wounded in 5 sites around the calyx. Each wound (2.5 wide and 3.4 mm depth) was first filled with 25 microl of a 0, 3.0, 6.0, 15.0, 30.0 or 60.0 mM 2-DG-water solution alone or combined with the yeast 8C at 10(8) cells/mL and then a 25 microl of the P. digitatum conidial suspension was added. Each treatment consisted of 3 replicates of 8 fruit (5 wounds/fruit) for a total 120 wounds per treatment. Oranges were maintained at 20 degrees C and high RH (95-98%) for up to 5 days, during which infection was monitored and the inhibitory activity calculated. The tests in vitro evedenced a significant slowing of the pathogen growth with the highest concentrations of 2-DG (15.0, 30.0 and 60.0 mM) with respect to the control; while at lower concentrations (1.5, 3.0, 6.0 mM) the development of the fungi was not significantly reduced. C. saitoana was resistant to all the doses employed to the abovementioned compound. In vivo the yeast alone was more effective compared to the sugar alone up to 6.0 mM while, at higher concentrations an additive effect was founded.