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Applied and Environmental Microbiology, February 2006, p. 1072-1078, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1072-1078.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Specific Detection of Xanthomonas axonopodis pv. dieffenbachiae in Anthurium (Anthurium andreanum) Tissues by Nested PCR{dagger}

Isabelle Robène-Soustrade,* Philippe Laurent, Lionel Gagnevin, Emmanuel Jouen, and Olivier Pruvost

UMR Peuplements Végétaux et Bioagresseurs en Milieu Tropical, CIRAD, Université de la Réunion, La Réunion, France

Received 3 December 2004/ Accepted 14 November 2005

Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires a sensitive and reliable diagnostic tool. A nested PCR test was developed from a sequence-characterized amplified region marker identified by randomly amplified polymorphic DNA PCR for the detection of X. axonopodis pv. dieffenbachiae. Serological and pathogenicity tests were performed concurrently with the nested PCR test with a large collection of X. axonopodis pv. dieffenbachiae strains that were isolated worldwide and are pathogenic to anthurium and/or other aroids. The internal primer pair directed amplification of the expected product (785 bp) for all 70 X. axonopodis pv. dieffenbachiae strains pathogenic to anthurium tested and for isolates originating from syngonium and not pathogenic to anthurium. This finding is consistent with previous studies which indicated that there is a high level of relatedness between strains from anthurium and strains from syngonium. Strains originating from the two host genera can be distinguished by restriction analysis of the amplification product. No amplification product was obtained with 98 strains of unrelated phytopathogenic bacteria or saprophytic bacteria from the anthurium phyllosphere, except for a weak signal obtained for one X. axonopodis pv. allii strain. Nevertheless, restriction enzyme analysis permitted the two pathovars to be distinguished. The detection threshold obtained with pure cultures or plant extracts (103 CFU ml–1) allowed detection of the pathogen from symptomless contaminated plants. This test could be a useful diagnostic tool for screening propagation stock plant material and for monitoring international movement of X. axonopodis pv. dieffenbachiae.

* Corresponding author. Mailing address: UMR Peuplements Végétaux et Bioagresseurs en Milieu Tropical, Pôle de Protection des Plantes, 7, chemin de l'Irat, 97410 Saint-Pierre, Réunion Island, France. Phone: (262) 262499277. Fax: (262) 262499293. E-mail: isabelle.soustrade{at}cirad.fr.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, February 2006, p. 1072-1078, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1072-1078.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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