Rapid detection of Phaeocytostroma sacchari in sugarcane using conventional polymerase chain reaction

Abstract

Diseases caused by fungi associated with adverse weather conditions are one of the main causes of decreases in sugarcane production. This study aimed to develop a protocol for a fast identification of Phaeocytostroma sacchari, which is the causal agent of bark rot in sugarcane. The reference sequences of three DNA regions of P. sacchari, namely internal transcribed spacer, ribosomal large subunit and translation elongation factor 1-alpha (TEFl-α), were analyzed with specific primers design. The specific primers generated that aligned in their entirety with P. sacchari were selected and synthesized. Polymerase chain reaction (PCR) assays were performed to confirm the primer specificity, using P. sacchari isolates and 10 species of other genera. Two sets of primers that amplify the TEF-1α region (PsFl/Psfl and PsF2/PsR2) showed a high specificity and sensitivity in detecting P. sacchari using conventional PCR, what will allow large-scale surveys of this pathogen in sugarcane crops.