Real time polymerase chain reaction for rapid and quantitative determination of Cystofilobasidium infirmominiatum on the surfaces of apple,pear, and sweet cherry fruit

The objectives of this study were to develop primers and a real time PCR protocol for the postharvest biocontrol yeast Cystofilobasidium infirmominiatum (Cim). The application of this technology was developed to quantify Cim on the surfaces of apple, two pear cultivars, and sweet cherry fruit treated over a range of concentrations. Statistically significant relationships were observed between Cim DNA on fruit surfaces, expressed as μg/m², and CFU/L of dip suspensions for apple, pear, and sweet cherry. In addition, the relationship for each fruit was significantly different from the other three fruits. Threshold values of concentrations of Cim DNA on the fruit surface were calculated based on regression equations and a dose of 2.0 × 10¹¹ CFU/L of dip suspension, the dose for optimum decay control, and were 4.8, 7.0, 16.5, and 25.2 μg/m² for Bosc pear, Lapins sweet cherry, d’Anjou pear, and Golden Delicious apple, respectively. Monitoring Cim DNA concentration on fruit surfaces will assure that Cim is being properly applied to fruit and that a sufficient number of cells are present for optimum decay control.