Real-time PCR assay to detect Verticillium albo-atrum and V. dahliae in hops: development and comparison with a standard PCR method.

Verticillium wilt, caused by Verticillium albo-atrum and V. dahliae, is a devastating disease that causes considerable economic crop losses in hop fields. The fungus can survive in soil for several years by producing resting structures, and due to the lack of effective fungicides, the disease is spreading. Thus, a fast and sensitive detection system is urgently needed. In this study we report a novel routine detection method for the identification of V. albo-atrum and V. dahliae on a molecular basis. The standard polymerase chain reaction (PCR) assay includes isolation of the fungus from affected tissue by following DNA extraction from fungal cultures and PCR identification by specific primers. In order to improve this detection assay, DNA was isolated directly from the hop bine using a commercially available DNA isolation kit. A multiplex real-time PCR assay for the simultaneous detection of V. albo-atrum and V …