. Fungal isolation and purification
The apple fruits were divided into species. The samples, which were apparently diseased, were cut from the advancing edges of the le- sion with a sterilized knife. The cut portion of the lesion was disin- fected with ethanol of 85% concentration for 2 minutes. These were then rinsed in three different changes of distilled water. Each portion was then homogenized using a sterile glass rod and a test tube containing 10 ml of the homogenate (1 g + 9 ml) (101) was made and serially diluted down to 104. Plates of already prepared Potato Dextrose Agar (PDA) containing Chloramphenicol (30 mg/l) to prevent the growth of bacteria were inoculated with
0.1 ml aliquots of the serially diluted samples and incubated at ambient room temperature (25–30 C) for seven days. After seven days, growth of fungal colonies on PDA was counted and recorded in a colony forming unit per milliliter (cfu/ml). Isolated species were sent for molecular confirmation.