Abstract
Extracellular Vesicles (EVs) play pivotal roles in cell-to-cell and inter-kingdom communication. Despite their relevant biological implications, the existence and role of plant EVs released into the environment has been unexplored. Herein, we purified round-shaped small vesicles (EVs) by differential ultracentrifugation of a sampling solution containing root exudates of hydroponically grown tomato plants. Biophysical analyses, by means of dynamic light scattering, microfluidic resistive pulse sensing and scanning electron microscopy, showed that the size of root-released EVs range in the nanometric scale (50–100 nm). Shot-gun proteomics of tomato EVs identified 179 unique proteins, several of which are known to be involved in plant-microbe interactions. In addition, the application of root-released EVs induced a significant inhibition of spore germination and of germination tube development of the plant pathogens Fusarium oxysporum, Botrytis cinerea and Alternaria alternata. Interestingly, these EVs contain several proteins involved in plant defense, suggesting that they could be new components of the plant innate immune system.
Highlights
Cells from all kingdoms of life secrete biomembrane-enclosed vesicles into the extracellular space.Extracellular vesicles (EVs) are a mix of heterogeneous populations of structures with a broad size distribution (30–1000 nm)
Based on the hypothesis that contaminating protein in the samples should have a lowering effect on the particles-to-protein ratio [17], the high particles-to-protein ratios measured in our samples could indicate that the vesicle preparation was pure or contained relatively less proteins when compared to mammalian systems
2020, 9, were isolated from a sampling solution of root exudates in ultrapure water of tomato (Solanum lycopersicum L.) plants cultivated in a hydroponic system, using membrane filtration
Summary
Cells from all kingdoms of life secrete biomembrane-enclosed vesicles into the extracellular space.Extracellular vesicles (EVs) are a mix of heterogeneous populations of structures with a broad size distribution (30–1000 nm). Several lines of evidence indicate that EVs are involved in the horizontal transfer of bioactive cargos, such as proteins, nucleic acids (DNA, mRNA and microRNA), and other molecules including carbohydrates, lipids and secondary metabolites [1]. Paramural vesicle structures between the plasma membrane and cell wall in plant tissues were firstly observed by transmission electron microscopy (TEM) more than half a century ago [3]. Only recently, they have been isolated from apoplastic washing fluids of Arabidopsis leaves and sunflower seeds [4,5]. Plant apoplastic vesicles have been proved to carry small non-coding RNAs, such as micro (miRNAs), short interfering RNAs (siRNAs) and a new class of “tiny
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