Skipping the Insect Vector: Plant Stolon Transmission of the Phytopathogen ‘Ca. Phlomobacter fragariae’ from the Arsenophonus Clade of Insect Endosymbionts

SIMPLE SUMMARY: Numerous plant sap-feeding insects are vectors of plant-pathogenic bacteria that cause devastating crop diseases. Some of these bacteria had initially been insect endosymbionts that eventually evolved the capacity to survive in plants after being frequently transmitted to plants by their insect hosts during feeding. An example for this evolutionary transition is the bacterial symbiont ‘Candidatus Phlomobacter fragariae’ (hereafter Phlomobacter) of the planthopper Cixius wagneri. Upon transmission to strawberry plants by its insect vector, the bacterium accumulates in the plant phloem and causes Strawberry Marginal Chlorosis disease. Using quantitative PCR and transmission electron microscopy, we demonstrate an additional plant-to-plant transmission route: Phlomobacter can be transmitted from an infected plant to daughter plants through stolons, a specific type of stem from which daughter plants can develop. Our results show that Phlomobacter was abundant in stolons and was efficiently transmitted to daughter plants, which developed disease symptoms. Hence, Phlomobacter is not only able to survive in plants, but can even be transmitted to new plant generations, independently from its ancestral insect host. ABSTRACT: The genus Arsenophonus represents one of the most widespread clades of insect endosymbionts, including reproductive manipulators and bacteriocyte-associated primary endosymbionts. Two strains belonging to the Arsenophonus clade have been identified as insect-vectored plant pathogens of strawberry and sugar beet. The bacteria accumulate in the phloem of infected plants, ultimately causing leaf yellows and necrosis. These symbionts therefore represent excellent model systems to investigate the evolutionary transition from a purely insect-associated endosymbiont towards an insect-vectored phytopathogen. Using quantitative PCR and transmission electron microscopy, we demonstrate that ‘Candidatus Phlomobacter fragariae’, bacterial symbiont of the planthopper Cixius wagneri and the causative agent of Strawberry Marginal Chlorosis disease, can be transmitted from an infected strawberry plant to multiple daughter plants through stolons. Stolons are horizontally growing stems enabling the nutrient provisioning of daughter plants during their early growth phase. Our results show that Phlomobacter was abundant in the phloem sieve elements of stolons and was efficiently transmitted to daughter plants, which rapidly developed disease symptoms. From an evolutionary perspective, Phlomobacter is, therefore, not only able to survive within the plant after transmission by the insect vector, but can even be transmitted to new plant generations, independently from its ancestral insect host.