AbstractThis study considers the influence of temperature, incubation time and virulence in the production of the toxin cercosporin by Cercospora coffeicola, the causal agent of brown eye spot in coffee. The area under the progress curve of cercosporin production (AUPCCP) was also evaluated. A pathogenicity test was performed in a group with 58 isolates of C. coffeicola, which allowed the selection of four isolates, two representing the highest (LFP 12 and LFP 59) and two the lowest (LFP 24 and LFP 43) severity groups. The four isolates were cultivated in potato dextrose agar (PDA) and incubated in growth chambers at different temperatures (15, 20, 25 and 30°C) for 20 days. The experiments were conducted in a randomized complete block design with four replications. The cercosporin production decreased in all tested isolates at 30°C, resulting in a lower AUPCCP. The higher values of cercosporin were obtained at 20 and 25°C between 8 and 12 days of incubation. The isolates that produced the highest and lowest cercosporin concentrations were LFP 12 and LFP 43, respectively. After 15 days of incubation, toxin production is practically null in all isolates independently of the incubation temperature. Thus, the hypothesis for quantification of cercosporin production as a variability parameter within the species is suggested.