Development of an engineered bioluminescent reporter phage for detection of bacterial blight of crucifers

David A. Schofield, Carolee Theresa Bull, Isael Rubio, W. Patrick Wechter, Caroline Westwater, Ian J. Molineux

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Bacterial blight, caused by the phytopathogen Pseudomonas cannabina pv. alisalensis, is an emerging disease afflicting important members of the Brassicaceae family. The disease is often misdiagnosed as pepper spot, a much less severe disease caused by the related pathogen Pseudomonas 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 bacterial luxAB genes encoding luciferase into the genome of P. cannabina pv. alisalensis phage PBSPCA1. The PBSPCA1::luxAB reporter phage is viable and stable and retains properties similar to those of the wildtype phage. PBSPCA1::luxAB rapidly and sensitively detects P. cannabina pv. alisalensis by conferring a bioluminescent signal response to cultured cells. Detection is dependent on cell viability. Other bacterial pathogens of Brassica species such as P. syringae pv. maculicola, Pseudomonas marginalis, Pectobacterium carotovorum, Xanthomonas campestris pv. campestris, and X. campestris pv. raphani either do not produce a response or produce significantly attenuated signals with the reporter phage. Importantly, the reporter phage detects P. cannabina pv. alisalensis on diseased plant specimens, indicating its potential for disease diagnosis.

Original languageEnglish (US)
Pages (from-to)3592-3598
Number of pages7
JournalApplied and environmental microbiology
Volume78
Issue number10
DOIs
StatePublished - May 1 2012

Fingerprint

Brassicaceae
blight
bacteriophages
Bacteriophages
pathogen
Pseudomonas syringae pv. maculicola
Xanthomonas campestris
Pseudomonas syringae
Pseudomonas
pathogens
Pseudomonas marginalis
Pectobacterium carotovorum
Xanthomonas campestris pv. campestris
viability
genome
Plant Diseases
Bacterial Genes
emerging diseases
Brassica
disease diagnosis

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Schofield, David A. ; Bull, Carolee Theresa ; Rubio, Isael ; Wechter, W. Patrick ; Westwater, Caroline ; Molineux, Ian J. / Development of an engineered bioluminescent reporter phage for detection of bacterial blight of crucifers. In: Applied and environmental microbiology. 2012 ; Vol. 78, No. 10. pp. 3592-3598.
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Development of an engineered bioluminescent reporter phage for detection of bacterial blight of crucifers. / Schofield, David A.; Bull, Carolee Theresa; Rubio, Isael; Wechter, W. Patrick; Westwater, Caroline; Molineux, Ian J.

In: Applied and environmental microbiology, Vol. 78, No. 10, 01.05.2012, p. 3592-3598.

Research output: Contribution to journalArticle

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