Nine fungi were isolated from the sugarcane setts from locations in Queensland and New South Wales cane-growing areas. The fungal isolates were grown on potato dextrose agar (PDA), indexed and maintained at Sugar Research Australia Woodford for further study. Growth characteristics and sporulation of the fungal isolates were recorded. A glasshouse experiment was conducted to determine the pathogenicity of the fungal isolates. The experiment was established at the end of July 2019 with nine isolates, one un-inoculated and one known fungal species (Athelia rolfsii, WF001). The fungal isolates in PDA were cut into small pieces and mixed with moist vermiculite. Sugarcane variety Q208A was collected from the Woodford propagation plots, cut into one-eye-setts (OES) and hot-water treated at 50.5°C for 30 minutes to eliminate internal systemic infection. The setts were then left on a bench inside a shed for 1 hour to dry before they were planted into the inoculated vermiculate trays, and maintained in a germination chamber at 30°C. Three vermiculite trays of each isolate with 10 OES in each tray were planted in a randomised complete-block design and, germination data were collected at the first, second, third and fourth weeks after planting. Morphological study showed that five isolates (WF 006, WF 008, WF 009, WF 011 and WF 012) were Colletotrichum spp., two isolates were Athelia rolfsii (WF 005 and WF 010) and two isolates (WF 007 and WF 013) yet to be identified. None of the isolates produced fruiting bodies or structure under ambient condition. Germination of setts were reduced in the trays inoculated with WF 010, WF 012, WF 007 and WF 001 when compared with the un-inoculated control. Other fungal isolates did not affect the sett germination. Further research is underway to identify more isolates and to determine their impact on sett germination.