Huge thanks to the panel members:

Prof. Carmen Montiel
Prof. Fernando Vivanco
Dr. Gloria Alvarez-Llamas
Dr. Milagros Castellanos
Prof. Luis Gandía

We provide evidence supporting the potential of the following protein families as additional PEV markers: aquaporins, vacuolar-type ATPase complex subunits, fasciclin-like arabinogalactan proteins FLA 10 and FLA13, syntaxins other than PEN1, germin-like proteins and calreticulins

Our data shows consistent specificity for current PEV markers from A. thaliana TET8 and PEN1, but these are not robustly in PEVs from other species. Alternative markers need to be explored, aiming to create a broader set of standard PEV identity markers

Next, we searched for the appearance of protein families from potential plant EV markers in previous proteomic studies of plant EVs. Conservation analyses throughout the plant and animal taxonomy of representative members of these families were also performed

We also analysed protein content of SEC fractions not pooled for isolating EVs and calculated an enrichment degree in EVs. This analysis offered key results to determine marker´s specificity, with some potential EV markers equally present in EVs and the soluble AWF proteome

To accomplish this, we have employed a two-step solubilization method to extract proteins from plant EVs for LC-MS/MS analysis, identifying as many as 1145 proteins in A. thaliana and 899 in B. oleracea, detected in all 3 biological replicates assayed

We achieved the most profound proteomic analysis of AWF-EVs (genuine EVs) from Arabidopsis and red cabbage, ranking potential markers by enrichment in EVs, species-specific abundance and plant specificity, also comparing with previously data and performing conservation analyses

There is urgent need for standardisation of identity markers and evidence of their conservation throughout plant taxa. Hoping this publication helps our efforts in the ISEV Plant Task Force building steps towards rigorous characterisation complying with MISEV.
Major findings👇