A minisatellite-based MLVA for deciphering the global epidemiology of the bacterial cassava pathogen Xanthomonas phaseoli pv. manihotis

PLoS One. 2023 May 11;18(5):e0285491. doi: 10.1371/journal.pone.0285491. eCollection 2023.

Abstract

Cassava Bacterial Blight (CBB) is a destructive disease widely distributed in the different areas where this crop is grown. Populations studies have been performed at local and national scales revealing a geographical genetic structure with temporal variations. A global epidemiology analysis of its causal agent Xanthomonas phaseoli pv. manihotis (Xpm) is needed to better understand the expansion of the disease for improving the monitoring of CBB. We targeted new tandem repeat (TR) loci with large repeat units, i.e. minisatellites, that we multiplexed in a scheme of Multi-Locus Variable number of TR Analysis (MLVA-8). This genotyping scheme separated 31 multilocus haplotypes in three clusters of single-locus variants and a singleton within a worldwide collection of 93 Xpm strains isolated over a period of fifty years. The major MLVA-8 cluster 1 grouped strains originating from all countries, except the unique Chinese strain. On the contrary, all the Xpm strains genotyped using the previously developed MLVA-14 microsatellite scheme were separated as unique haplotypes. We further propose an MLVA-12 scheme which takes advantage of combining TR loci with different mutation rates: the eight minisatellites and four faster evolving microsatellite markers, for global epidemiological surveillance. This MLVA-12 scheme identified 78 haplotypes and separated most of the strains in groups of double-locus variants (DLV) supporting some phylogenetic relationships. DLV groups were subdivided into closely related clusters of strains most often sharing the same geographical origin and isolated over a short period, supporting epidemiological relationships. The main MLVA-12 DLV group#1 was composed by strains from South America and all the African strains. The MLVA-12 scheme combining both minisatellite and microsatellite loci with different discriminatory power is expected to increase the accuracy of the phylogenetic signal and to minimize the homoplasy effects. Further investigation of the global epidemiology of Xpm will be helpful for a better control of CBB worldwide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Typing Techniques
  • Genotype
  • Manihot* / genetics
  • Microsatellite Repeats / genetics
  • Minisatellite Repeats* / genetics
  • Phylogeny

Supplementary concepts

  • Xanthomonas phaseoli

Grants and funding

We thank the Faculty of Sciences from Universidad de los Andes-Bogotá, Colombia (INV-2021-128-2283), the UMR Interactions Plantes Microorganismes Environnement, and the Agropolis Foundation (project PAIX, grant no. 1 403-073), Montpellier, France, for financial support. The Ecos Nord programme supported inter-laboratory mobility (grant no. C15A01). Leidy Rache was supported by Colciencias with a doctoral fellowship (call No. 528, 2011). Olivier Pruvost acknowledges the European Union (ERDF contract GURDT I2016‐1731‐0006632) and Réunion regional council for support.The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript