Item Details

Title: Multiplex PCR for specific and robust detection of Xanthomonas campestris pv. musacearum in pure culture and infected plant material

Date Published: 2011
Author/s: J. Adriko, V. Aritua, C. N. Mortensen, W. K. Tushemereirwe, J. Kubiribab and O. S. Lund
Data publication:
Funding Agency : DANIDA under the ENRECA project LIFE – 731
Copyright/patents/trade marks: British Society for Plant Pathology, BSPP
Journal Publisher: Plant Pathology
Affiliation: Danish Seed Health Centre for Developing Countries, Department of Agriculture and Ecology, Faculty of Life Sciences, University of
Copenhagen, Hoejbakkegaard Alle 30 DK-2630 Taastrup, Denmark; bNational Banana Research Programme, National Agricultural
Research Laboratories, PO Box 7065, Kampala, Uganda; and cCitrus Research and Education Center, University of Florida, 700
Experiment Station Road, Lake Alfred, FL 33850, USA
Keywords: Banana, general secretion pathway protein D, multiplex PCR, specific disease diagnosis, Xanthomonas campestris pv. musacearum


The present study developed a pathovar-specific PCR for the detection of Xanthomonas campestris pv. musacearum (Xcm),
the cause of banana xanthomonas wilt, by amplification of a 265-bp region of the gene encoding the general secretion
pathway protein D (GspD). A distinct DNA fragment of the expected size was amplified from genomic DNA from all of 12
Xcm isolates tested and no amplification of DNA was observed from other xanthomonads or plant-associated bacteria,
including the two closely related species Xanthomonas vasicola pv. holcicola and Xanthomonas axonopodis pv. vasculorum.
The Xcm-specific PCR was successfully multiplexed with internal control primers targeting 16S rDNA for application
on DNA from bacterial cultures and with primers targeting plant mitochondrial 26S rDNA for application on DNA
extracted from plant material. Diagnostic discrimination of healthy and infected plants was subsequently demonstrated in
tests on artificially inoculated screenhouse cultivars of banana and field bananas with and without symptoms sampled from
different parts of Uganda. This study therefore demonstrated a robust and specific Xcm diagnostic tool with the added
advantage of applying internal PCR controls for direct quality assessment of results.