Exosomes in the phloem and xylem of woody plants.

Exosomes in the secondary phloem and secondary xylem of angiosperms and gymnosperms have physiological roles in the storage and transport of endoglucanases. Knowledge of plant extracellular vesicles (EVs) is limited by their presence in the apoplastic fluid of seeds and leaves. The contents of plant EVs and their biological functions are unclear. The aim of the present study was to expand our knowledge of EVs in woody plants. Sample splits were prepared from branch and stem samples from angiosperms and gymnosperms after cryomechanical destruction with liquid nitrogen. The study methods included scanning electron (SEM), atomic force microscopy (AFM), endoglucanase activity measurement. EVs visualized on the internal layers of the cell walls proved to be exosomes according to their diameter (65-145 nm). SEM revealed cup-shaped structures characteristic of exosomes in a dry state. Plant exosomes in the form of globules in the native state were visualized for the first time by AFM. Exosomes were present both in the active and dormant cambium. Erosion zones were observed at the sites of exosome localization. The activity of endo-1,4-β-glucanase was detected in Picea xylem, while the RNA level was very low, suggesting that endo-1,4-β-glucanases were preserved in the exosomes. There are grounds to assert that endo-1,4-β-glucanases delivered by exosomes participated in pit cavity formation in the S layer of xylary fibres. A possible mechanism of endo-1,4-β-glucanase action in the biosynthesis of the secondary wall is proposed. These results demonstrate that the physiological role of the exosomes in the phloem and xylem is the storage and transport of endo-1,4-β-glucanases participating in cell wall remodeling in woody plants. Present study expands our knowledge about plant exosomes.