Tandem repeats as genotyping tools for understanding the genetic structure and epidemiology of Xanthomonas citri pv. citri

MultiLocus Variable number of tandem repeat Analysis (MLVA) has been extensively used to examine epidemiological and evolutionary issues on monomorphic human pathogenic bacteria. MLVA is gaining popularity on agriculturally important bacterial plant pathogens as a tool to improve our understanding of their epidemiology. Xanthomonas citri pv. citri, the causal agent of Asiatic citrus canker, is a quarantine organism in several countries and a major threat for the citrus industry worldwide. We screened the genomes of X. citri pv. citri strain IAPAR 306 and of phylogenetically related strains for tandem repeats. Two MLVA schemes targeting minisatellites and microsatellites, respectively, were developed to assess the diversity of this monomorphic bacterium at various spatio-temporal scales. Microsatellites are useful for outbreak investigation. In contrast, minisatellites were used to decipher the global diversity of X. citri pv. citri and revealed four pathotype-specific lineages based on Discriminant Analysis of Principal Components (DAPC). A single lineage (DAPC1) comprised strains that were implicated in the major geographical expansion of X. citri pv. citri during the 20th century. When applied to the analysis of the emergence of in Africa, minisatellites revealed the presence of two different genetic lineages in addition to DAPC1 on this continent, consistent with SNP data derived from draft genome sequences. Using bacterial populations collected in two neighboring Western African countries, Mali and Burkina Faso, we applied these two genotyping schemes to investigate the origin and pathways of these emergences. Minisatellite and SNP data suggested the introduction of two groups of strains in Mali (DAPC1 and DAPC2). DAPC2 was restricted to Bamako district, whereas DAPC1 strains were found much more invasive. The latter strains formed a major clonal complex based on microsatellite data. This suggests that human activities played a major role in the spread of DAPC1 strains via the movement of contaminated propagative material, further supported by the low differentiation between populations from geographically distant nurseries and orchards and the fact that many nursery strains were assigned to the primary or secondary founder haplotypes identified in the minimum spanning tree. Approximate Bayesian Computation analyses supported the hypothesis that strains from Burkina Faso resulted from a bridgehead invasion from Mali. The new minisatellite scheme represents an opportunity for international X. citri pv. citri genotyping and data sharing. The data generated in this study was deposited in the Xanthomonas citri genotyping database (http://www.biopred.net/MLVA/). (Texte intégral