A PCR-based assay for specific detection of Acremonium implicatum, an endophytic fungus in species of Brachiaria
Kelemu, S.; Dongyi, H.; Guixiu, H.; Takayama, Y.
Phytopathology 92(6 Suppl): S41
Brachiaria is a pan-tropical genus with about 100 species. Some of the African species are economically important forage grasses in tropical America. An endophytic association of Acremonium implicatum with species of Brachiaria has been identified. DNA from isolates of A. implicatum was amplified with arbitrary 10-mer primers. A 500-bp polymerase chain reaction (PCR) product amplified with primer OPAK10 and common to most of the isolates of A. implicatum was cloned, and sequenced. Two specific primers, P1 (5'-TTCGAATGATAAGGCAGATC-3') and P4 (5'-ACGCATCCACTGTATGCTAC-3') were synthesized. The primer pair amplifies a single fragment of about 450-bp from DNA of isolates of A. implicatum whether from pure culture or in association with Brachiaria plants. No amplification product was detected using DNA from endophyte-free plants or from pathogenic and non-pathogenic fungi associated with Brachiaria. This assay allows precise and rapid detection of endophytes in Brachiaria plants and permits differentiation between endophytic and non-endophytic fungi.