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Poussier, S.

Publications and source records attributed to Poussier, S..

4 recordsLinked to original sources

Ecology, genomics and biocontrol potential of bacteriophages infecting the bacterial wilt pathogen Ralstonia solanacearum species complex in Reunion Island

The Ralstonia solanacearum species complex (RSSC), the causal agent of bacterial wilt, is among the most destructive soil-borne plant pathogens worldwide, yet effective and sustainable control strategies remain limited. Bacteriophages represent promising biocontrol agents, but their efficacy depends on ecological compatibility with local pathogen populations. Here, we combined ecological sampling, comparative genomics, phenotypic characterization and plant assays to investigate RSSC-infecting phages in Reunion Island and evaluate their biocontrol potential. We isolated 45 phages from diverse agricultural matrices and obtained complete genome sequences for 35 novel isolates. Phylogenomic analyses revealed a locally diversified assemblage comprising multiple known taxa and several putative new genera, forming clusters largely distinct from global reference phages. Phage diversity and antibacterial activity were structured primarily by bacterial phylogeny rather than plant host or geographic origin, indicating that plants act mainly as ecological interfaces while environmental bacterial populations shape phage specialization. The community displayed two contrasting evolutionary strategies: expanding virulent lineages associated with strong antibacterial activity and persistent temperate lineages carrying integration and host-interaction functions. Host-range assays confirmed phylotype-dependent susceptibility, and strictly lytic phages showed consistently higher inhibitory activity. Guided by combined genomic and phenotypic screening, we designed a multi-family phage cocktail targeting dominant local RSSC lineages. The cocktail exhibited strong in vitro suppression of bacterial growth and significantly reduced disease severity in tomato plants. Together, our results demonstrate that effective phage biocontrol depends on evolutionary matching between phages and regional pathogen populations. Integrating ecological, genomic and functional characterization provides a robust framework for selecting locally adapted phages and developing durable phage-based strategies for managing bacterial wilt.

microbiology↗

Robust design and validation of LAMP assays for in-field detection of three major bacterial vascular diseases of banana

Bacterial diseases of banana are becoming increasingly significant worldwide, resulting in reduced yields and higher disease management costs. The most important bacterial diseases of banana include Moko and banana blood disease (BBD), caused by Ralstonia solanacearum and Ralstonia syzygii subsp. celebesensis, respectively, and banana Xanthomonas wilt (BXW) caused by Xanthomonas vasicola pv. musacearum. Effective surveillance and disease management require point-of-care diagnostics, such as loop-mediated isothermal amplification (LAMP), for on-site operation. In this study, three LAMP assays were developed to specifically detect the bacteria responsible for Moko, BBD and BXW, directly from banana tissues, using a simplified DNA extraction protocol. The BBD - and BXW-LAMP assays demonstrated 100% specificity, yielding negative results for a broad range of non-target bacteria, including closely related species as well as pathogenic and endophytic strains associated with banana, and positive results for all the tested target strains. For Moko disease, a duplex-LAMP assay was developed to detect all strains from the four globally most relevant sequevars: IIB-3, IIB-4, IIA-6, and IIA-24. The duplex-LAMP successfully detected all target strains, except one that was shown to be non-pathogenic to Cavendish bananas. All non-target strains tested negative, with the exception of a delayed signal for one strain belonging to Ralstonia thomasi, not associated with banana environment (hospital strain). These results were supported by an extensive in silico analysis conducted on 9,668 Burkholderiaceae and 7,483 Xanthomonadaceae genomes. Detection limits ranged from 0.1 pg/{micro}l to 1 pg/{micro}l DNA, and from 10 to 10 CFU/ml on banana tissues spiked with calibrated bacterial suspensions, depending on the assay. The LAMP assays prove highly effective for detecting target pathogens in both artificially inoculated banana plants and field samples, offering a promising tool for improving disease management strategies.

molecular biology↗

Emergence and host range expansion of an epidemic lineage of Ralstonia solanacearum

The evolutionary processes underlying disease outbreaks remain unknown for most bacterial plant pathogens. We sequenced an outbreak of lethal wilt disease in Martinique caused by two distantly related lineages in the Ralstonia solanacearum species complex. One lineage (R. solanacearum IIB-4NPB) exhibited the broadest host range ever documented for a single lineage in the field, while the other (R. pseudosolanacearum I-18) expanded in parallel but retained specialisation on solanaceaous hosts. Phylogenomic analysis of 407 outbreak isolates shows both lineages independently disseminated from mainland populations into Martinique. We resolved fine-scale geographic patterns of genomic diversity and identified spatial hotspots of interspecies mobile element exchange, resulting in the identification of a new family of Ralstonia integrative conjugative elements (ICEs) associated with the outbreak. ICE accessory gene integration sites display striking functional specialization and differentiation despite variable gene content: each site acquires only metabolism-associated or defence-related genes, respectively. This work provides insight into the origin and genomic changes associated with an outbreak of plant disease, and highlights the role of mobile elements in driving pathogen emergence.

microbiology↗

A phylogenetic host range index reveals contrasted relationships between phage virulence and specialisation

Phages are typically known for having a limited host range, targeting various strains within a specific bacterial species. However, factors like the phylogeny or epidemiology of host bacteria are often disregarded, despite their potential influence on phage specialization and virulence. This research utilizes a new "phylogenetic host range index" that accounts for the genetic diversity of bacterial hosts, to classify phages into specialists and generalists accurately. We provide evidence that the CRISPR-Cas immune system of bacteria more frequently targets generalist phages than specialist phages. We explore the hypothesis that generalist phages might exhibit lower virulence than specialist ones due to potential evolutionary trade-offs between host range breadth and virulence. Importantly, contrasted correlations between phage virulence and host range depend on the epidemiological context. A trade-off was confirmed in a homogeneous bacterial epidemiology situation, but not in more complex epidemiological scenario, where no apparent costs were detected for phages adapted to a wide range of hosts. This study highlights the need for genetic analyses in phage host range and of investigating ecological trade-offs that could improve their applications in biocontrol or therapy.

microbiology↗