Diagnosis of "maladie des feuilles cassantes" or brittle leaf disease of date palms by detection of associated chloroplast encoded double stranded RNAs

The “Maladie des feuilles cassantes” (MFC) or “Brittle leaf disease” of date palms is associated with the accumulation of two populations of small, chloroplast-encoded RNAs. A plasmid vector containing a cDNA with partial sequences of both of these RNA populations was used to synthesize a DIG-labeled bifunctional probe by PCR. The probe has been tested to detect, by molecular hybridization, MFC-associated RNAs from dsRNA-enriched palm leaflet preparations. Leaflet samples from MFC-affected date palm trees consistently gave a positive hybridization signal regardless of the date palm cultivar, severity of symptoms, or geographical location, whereas samples from date palm trees affected by other biotic and abiotic stresses tested negative. The assay is specific for MFC and can be used for early diagnostic purposes

A bacterial endosymbiont of the fungus Rhizopus microsporus drives phagocyte evasion and opportunistic virulence

This is the final version. Available on open access from Elsevier via the DOI in this recordData and code availability: Genomic sequencing data is available through the Sequence Read Archive (SRA) on NCBI and accession codes for sequences referenced in this work are: R. pickettii: SRA: SRR16523121 and R. microsporus: SRA: SRR16610381. This work did not lead to the generation of any new code.Opportunistic infections by environmental fungi are a growing clinical problem, driven by an increasing population of people with immunocompromising conditions. Spores of the Mucorales order are ubiquitous in the environment but can also cause acute invasive infections in humans through germination and evasion of the mammalian host immune system. How they achieve this and the evolutionary drivers underlying the acquisition of virulence mechanisms are poorly understood. Here, we show that a clinical isolate of Rhizopus microsporus contains a Ralstonia pickettii bacterial endosymbiont required for virulence in both zebrafish and mice and that this endosymbiosis enables the secretion of factors that potently suppress growth of the soil amoeba Dictyostelium discoideum, as well as their ability to engulf and kill other microbes. As amoebas are natural environmental predators of both bacteria and fungi, we propose that this tri-kingdom interaction contributes to establishing endosymbiosis and the acquisition of anti-phagocyte activity. Importantly, we show that this activity also protects fungal spores from phagocytosis and clearance by human macrophages, and endosymbiont removal renders the fungal spores avirulent in vivo. Together, these findings describe a new role for a bacterial endosymbiont in Rhizopus microsporus pathogenesis in animals and suggest a mechanism of virulence acquisition through environmental interactions with amoebas.Wellcome TrustBiotechnology and Biological Sciences Research Council (BBSRC)Darwin Trust of EdinburghNational Institutes of Allergy and ImmunologyMedical Research Council (MRC)Royal Societ