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Published on: July 15, 2011
RAPD-PCR Fingerprinting and Southern Analysis of Xanthomonas axonopodis pv. dieffenbachiae Strains Isolated from
M H R Khoodoo1, Y Jaufeerally-Fakim1
1Biotechnology Unit, Faculty of Agriculture, University of Mauritius, Réduit, Mauritius.
Abstract:
Anthurium blight, caused by Xanthomonas axonopodis pv. dieffenbachiae, is a systemic disease of Anthurium and other aroids. The aims of this work were to study the genetic diversity among X. axonopodis pv. dieffenbachiae strains and to identify, from the polymerase chain reaction (PCR) profiles, DNA probes that would be specific for the pathovar dieffenbachiae. Twenty-five X. axonopodis pv. dieffenbachiae strains, isolated from different hosts and geographical locations including Mauritius, were fingerprinted using the random amplified polymorphic DNA (RAPD)-PCR technique. The fingerprints were analyzed by the National Taxonomy System Software (NTSYS). The specificity of some of the RAPD fragments selected from PCR profiles was tested by Southern analyses of the PCR products. Ten arbitrary primers were chosen from an initial set of 111 decamers. Two hundred and nine RAPD markers were generated in eight individual DNA profiles. A correlation was found between the serotypes and the RAPD profiles for some groups of isolates. A possible link was also observed between the host range of the isolates tested and their RAPD profiles for strains isolated from Dieffenbachia and Philodendron. These results were confirmed by Southern analysis. Cluster analysis by the unweighted pair group method, arithmetic average (UPGMA) confirmed that the pathovar is genetically diverse with some strains that were clustered together showing similar host preferences. DNA probes with a potential use in molecular diagnostics of Anthurium blight were identified. This preliminary work could be used to develop PCR primers that will enable the sensitive detection of the pathogen in latently infected plants.
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