Development of a Real-Time PCR Assay to Detect the Kiwifruit Pathogen

Pectobacterium actinidiae is a significant kiwifruit canker pathogen reported from kiwifruit-growing areas in Asia and South America. To improve the diagnostic capability for this bacterium, a probe-based real-time PCR assay was developed for sensitive and fast detection of P. actinidiae. The highly specific assay targets the meta-pathway phenol degradation protein-like locus, unique to P. actinidiae. Validation showed that the assay is highly sensitive, detecting the pathogen down to 0.1 pg of pure genomic DNA or 10 to 50 colony-forming units in plant tissue matrix. To assess the competency of the tested DNA samples and avoid false-negative results, the assay was duplexed with plant internal control. This new method provides a valuable tool for specific and sensitive testing of imported and exported plant material to ensure freedom from P. actinidiae and prevent the spread of this pathogen into new areas.[Formula: see text]