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Systematics and Evolution |
Laboratoire de Phytopathologie, CIRAD-FLHOR, 97448 Saint-Pierre Cedex, La Réunion, France1
Laboratoire de Biologie Moléculaire des Relations Plantes-Microorganismes, INRA-CNRS, BP27, 31326 Castanet-Tolosan Cedex, France2
Unité de Biométrie et d’Intelligence Artificielle, INRA, BP27, 31326 Castanet-Tolosan Cedex, France3
Author for correspondence: Stéphane Poussier. Tel: +33 262 35 76 30. Fax: +33 262 35 76 41. e-mail: poussier{at}cirad.fr
The genetic diversity among strains in a worldwide collection of Ralstonia solanacearum, causal agent of bacterial wilt, was assessed by using three different molecular methods. PCR-RFLP analysis of the hrp gene region was extended from previous studies to include additional strains and showed that five amplicons were produced not only with all R. solanacearum strains but also with strains of the closely related bacteria Pseudomonas syzygii and the blood disease bacterium (BDB). However, the three bacterial taxa could be discriminated by specific restriction profiles. The PCR-RFLP clustering, which agreed with the biovar classification and the geographical origin of strains, was confirmed by AFLP. Moreover, AFLP permitted very fine discrimination between different isolates and was able to differentiate strains that were not distinguishable by PCR-RFLP. AFLP and PCR-RFLP analyses confirmed the results of previous investigations which split the species into two divisions, but revealed a further subdivision. This observation was further supported by 16S rRNA sequence data, which grouped biovar 1 strains originating from the southern part of Africa.
Keywords: bacterial wilt, PCR-RFLP, hrp, AFLP, 16S rRNA
Abbreviations: AFLP, amplified fragment length polymorphism; BDB, blood disease bacterium; HCA, hierarchical cluster analysis; UPGMA, unweighted pair group method with arithmetic averages
The GenBank accession numbers for the sequences determined in this work are AF207891–AF207897.
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