Wash test Fifty wheat seeds were added to 10 mL
sterile distilled water in a 100 mL flask. One drop of
sterile Tween 20 was added to the flask and then the
flask was vigorously shaken for 10 min. The suspension
was transferred to a 10 mL tube and centrifuged at
3 000 r/min for 5 min. The pallets were resuspended in
1 mL Shear’s solution and examined for Tilletia spp.
and Ustilago spp. with the help of a microscope (McGee
1988; Mathur and Cunfer 1993)