PCR-Mediated Detection of Sporisorium scitamineum in Sugarcane based on the bE Mating-Type Gene Sequence

Culmicolous smut caused by Sporisorium scitamineum is a main sugarcane disease, occurring in all sugarcane-producing areas of the world. The disease develops sori with whip-shaped axes corresponding to columellae and causes economic yield loss. In this study, a PCR-mediated assay was developed based on the b E mating-type gene sequence for the early detection of S. scitamineum in sugarcane tissues. Primer pair CLS 320 F/CLS 320R was designed and tested for the detection of S. scitamineum in sugarcane tissue. It was successful to rapidly detect S. scitamineum in DNA extracted from as few as 10 mg of diseased sugarcane tissue within one hour. The detection limit of the primer pair was tested and shown to amplify DNA at the pg level. The amplification products could be detected in DNA extracted from stem, whip-shaped axes and leaves of diseased plants. This high specificity assay can be applied to the detection of S. scitamineum in infected sugarcane tissues. Results presented here indicate that the primer pair CLS 320 F/CLS 320R is efficient in identifying S. scitamineum and accurately and rapidly detecting the disease caused by this pathogen in the field.