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The ascomycete fungus  Mycosphaerella polygoni - cuspidati  has been undergoing evaluation as a potential classical biological control agent for the invasive weed  Fallopia japonica  (Japanese knotweed), which has become troublesome in Europe and North America. In advance of the potential release of a biocontrol agent into a new environment, it is crucial to develop an effective monitoring system to enable the evaluation of agent establishment and dispersal within the target host population, as well as any potential attacks on non-target species. Therefore, a primer pair was designed for direct, rapid, and specific detection of the Japanese knotweed pathogen  M .  polygoni - cuspidati  based on the sequences of the internal transcribed spacer regions including the 5.8S rDNA. A PCR product of approximately 298 bp was obtained only when the DNA extracted from mycelial fragments of  M .  polygoni - cuspidati  was used. The lower limit of detection of the PCR method was 100 fg of genomic DNA. Using the specific primer pair,  M .  polygoni - cuspidati  could be detected from both naturally and artificially infected Japanese knotweed plants. No amplification was observed for other  Mycosphaerella  spp. or fungal endophytes isolated from  F. japonica . The designed primer pair is thus effective for the specific detection of  M .  polygoni - cuspidati  in planta.

Species-Specific Detection of Mycosphaerella polygoni-cuspidati as a Biological Control Agent for Fallopia japonica by PCR Assay