TY - JOUR T1 - Biophysical and proteomic analyses suggest functions of <em>Pseudomonas syringae</em> pv <em>tomato</em> DC3000 extracellular vesicles in bacterial growth during plant infection JF - bioRxiv DO - 10.1101/2021.02.08.430144 SP - 2021.02.08.430144 AU - Martin Janda AU - Christina Ludwig AU - Katarzyna Rybak AU - Chen Meng AU - Egidio Stigliano AU - Leon Botzenhardt AU - Beata Szulc AU - Jan Sklenar AU - Frank L.H. Menke AU - Jacob G. Malone AU - Andreas Brachmann AU - Andreas Klingl AU - Silke Robatzek Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/02/08/2021.02.08.430144.abstract N2 - Vesiculation is a process employed by Gram-negative bacteria to release extracellular vesicles (EVs) into the environment. Bacterial EVs contain molecular cargo from the donor bacterium and play important roles in bacterial survival and growth. Here, we describe EV production in plant-pathogenic Pseudomonas syringae pv. tomato DC3000 (Pto DC3000), the causal agent of bacterial speck disease. Cultured Pto DC3000 exhibited EV structures both on the cell surface and in the vicinity of bacterial cells, observed as outer membrane vesicle (OMV) release. We used in-solution trypsin digestion coupled to mass spectrometry to identify 369 proteins enriched in EVs recovered from cultured Pto DC3000. The predicted localization profile of EV proteins supports the production of EVs also in the form of outer-inner-membrane vesicles (OIMVs). EV production varied slightly between bacterial lifestyles and also occurred in planta. The potential contribution of EVs to Pto DC3000 plant infection was assessed using plant treatments and bioinformatic analysis of the EV-enriched proteins. While these results identify immunogenic activities of the EVs, they also point at roles for EVs in bacterial defences and nutrient acquisition by Pto DC3000.Competing Interest StatementThe authors have declared no competing interest. ER -