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Bacterial blight, caused by the phytopathogenPseudomonas cannabina pv.alisalensis , is an emerging disease afflicting important members of theBrassicaceae family. The disease is often misdiagnosed as pepper spot, a much less severe disease caused by the related pathogenPseudomonas syringae pv.maculicola . We have developed a phage-based diagnostic that can both identify and detect the causative agent of bacterial blight and differentiate the two pathogens. A recombinant “light”-tagged reporter phage was generated by integrating bacterialluxAB genes encoding luciferase into the genome ofP. cannabina pv.alisalensis phage PBSPCA1. The PBSPCA1::luxAB reporter phage is viable and stable and retains properties similar to those of the wild-type phage. PBSPCA1::luxAB rapidly and sensitively detectsP. cannabina pv.alisalensis by conferring a bioluminescent signal response to cultured cells. Detection is dependent on cell viability. Other bacterial pathogens ofBrassica species such asP. syringae pv.maculicola ,Pseudomonas marginalis ,Pectobacterium carotovorum ,Xanthomonas campestris pv.campestris , andX. campestris pv.raphani either do not produce a response or produce significantly attenuated signals with the reporter phage. Importantly, the reporter phage detectsP. cannabina pv.alisalensis on diseased plant specimens, indicating its potential for disease diagnosis.

Development of an Engineered Bioluminescent Reporter Phage for Detection of Bacterial Blight of Crucifers