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Detection and identification of the crucifer pathogen, Xanthomonas campestris pv. campestris, by PCR amplification of the conserved Hrp/type III secretion system gene hrcC

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Abstract

The development of a rapid detection method for Xanthomonas campestris pv. campestris (Xcc) in crucifer seeds and plants is essential for high-throughput certification purposes. Here we describe a diagnostic protocol for the identification/detection of Xcc by PCR amplification of fragments from the pathogenicity-associated gene hrcC. Under stringent conditions of amplification, a PCR product of 519 bp from hrcC was obtained from a collection of 46 isolates of Xcc, with the exception of two isolates from radish. No amplicons were obtained from 39 pure cultures of the phytopathogenic bacteria Xanthomonas campestris pv. cerealicola, X. campestris pv. juglandis, X. campestris pv. pelargonii, X. campestris pv. vitians, X. arboricola pv. pruni, X. axonopodis pv. phaseoli, X. axonopodis pv. vesicatoria, X. vesicatoria, Pseudomonas syringae pv. phaseolicola, P. syringae pv. syringae, P. syringae pv. tomato, P. fluorescens, P. marginalis, Pectobacterium atrosepticum, P. carotovorum subsp. carotovorum. In addition, PCR reactions were negative for fifty unidentified environmental isolates purified from the surface of crucifers. The PCR fragment was obtained from four strains previously classified as X. campestris pv. aberrans, X. campestris pv. armorociae, X. campestris pv. barbarae and X. campestris pv. incanae using pathogenicity assays. Our PCR protocol specifically detected Xcc in inoculated leaves, seeds and naturally infected leaves of crucifers.

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Acknowledgements

This study was supported with funds appropriated from the CNR (Italy). We would like also to thank Prof. Roberto Buonaurio (University of Perugia, Italy), Prof. Nicola Jacobellis (University of Basilicata, Italy), Prof. Astolfo Zoina (University of Napoli. Italy), Dr. Vittoria Catara (University of Catania, Italy), Dr. Stefania Loreti (C.R.A.-ISPaVe, Italy) for supplying some bacterial isolates used in this study.

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Authors and Affiliations

  1. C.R.A-Istituto Sperimentale per le Colture Industriali, SS 18 n° 204, Battipaglia, 84091, Salerno, Italy

    Massimo Zaccardelli, Francesco Campanile & Annalisa Spasiano

  2. Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA, 02115, USA

    Massimo Merighi

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  1. Massimo Zaccardelli
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  2. Francesco Campanile
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  3. Annalisa Spasiano
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  4. Massimo Merighi
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Correspondence to Massimo Zaccardelli.

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Zaccardelli, M., Campanile, F., Spasiano, A. et al. Detection and identification of the crucifer pathogen, Xanthomonas campestris pv. campestris, by PCR amplification of the conserved Hrp/type III secretion system gene hrcC . Eur J Plant Pathol 118, 299–306 (2007). https://doi.org/10.1007/s10658-007-9115-y

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