Detection and differentiation of Phaeoisariopsis griseola isolates with the polymerase chain reaction and group-specific primers

Specific detection of the two major groups of Phaeoisariopsis griseola (Andean and Mesoamerican) from infected common bean (Phaseolus vulgaris) leaves was achieved by amplification of different-sized DNA fragments with polymerase chain reaction (PCR) using group-specific primer pairs. These primer pairs were designed based on DNA sequences of cloned random amplified polymorphic DNA (RAPD) fragments. Using this method, P. griseola isolates from diverse geographical regions (five countries) were differentiated into the two previously established groups. Various sources of fungal tissue and DNA extraction methods were tested in order to develop a rapid PCR-based method to detect and differentiate P. griseola isolates. A simple and rapid sonication method was developed that allowed for PCR detection of P. griseola from mycelia or synnemata and conidia collected from angular leaf spot lesions on bean leaves.