| dc.description.abstract | The role of extracellular small RNA (sRNA) in interkingdom communication between
microorganisms and their eukaryotic hosts has been revealed recently. Extracellular vesicles
(EVs) are a significant delivery route of sRNAs, thus regulating gene expression in the recipient
cell. Such interactions have been shown between fungi and plants as well as bacteria and
animals but, little is known about sRNA-mediated host responses between bacteria and plants
or fungi.
This project aims to study the role of sRNAs in the outer membrane vesicles (OMVs) of the
plant growth promoting bacteria (PGPB) (of genera Paraburkholderia, Pseudomonas and
Enterobacter) in the naturally occurring communication with plant (Arabidopsis thaliana) and
phytopathogenic fungi (Botrytis cinerea). Further, we will explore the ability of engineered
OMV-sRNAs to manipulate fungal genes, as a means to suppress fungal diseases of plants.
To monitor the delivery of the OMV cargo into plant and fungi, OMVs and sRNAs will be
fluorescently labeled and OMVs’ interaction with recipient cells, and RNA delivery, will be
monitored by confocal microscopy. sRNA from OMVs and cells of bacteria, grown separately or
in the interaction with the plant or fungus, will be sequenced and compared to define the
criteria for the selection of sRNAs to be packed into OMVs. To identify sRNAs delivered by
OMVs to recipient cells, sRNA-Seq and bioinformatics analysis of plant and fungal cells upon
exposure to bacteria will be performed. The ability of OMV-sRNAs to modulate the expression
of predicted target genes in the host cells will be tested both for endogenous bacterial and
engineered sRNAs, designed to target recipient’s gene(s).
The main expected results: the visualization of OMV cargo delivery to recipient cells; revealing
the selection criteria for sRNA packaging into OMVs; and evidence of OMV-sRNAs mediated
regulation of host genes will be novel, ground-breaking discoveries, impacting many research
fields , from agriculture to medicine. The project will pave the way for using engineered OMVs-
sRNAs to modulate plant/fungal physiology, encouraging further studies of the application of
this process in biocontrol against phytopathogenic fungi, with a potential to its extrapolation to
animal and human fungal pathogens. | sr |
