{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T06:07:24Z","timestamp":1789106844529,"version":"build-2803163510"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001321","name":"National Research Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001321","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Plant Sci."],"abstract":"<jats:p>\n                    Extracellular vesicles (EVs) are lipid-bound structures that transport bioactive molecules and are increasingly recognized as key mediators of plant-pathogen interactions. Although EV secretion has been reported in filamentous pathogens, their roles in plant immunity, microbial interactions and cargo-sorting mechanisms remain poorly understood. This study aimed to characterize the functional roles and composition of EVs produced by\n                    <jats:italic>P. nicotianae<\/jats:italic>\n                    , and to investigate shared EV-associated protein categories and putative cargo-sorting signals, including KFERQ-like (Lys-Phe-Glu-Arg-Gln) motifs, across filamentous pathogens. EVs were isolated and characterized using transmission electron microscopy (TEM) and Nanoparticle tracking analysis (NTA). TEM confirmed the presence of membrane-bound vesicles with both single- and double-membrane morphologies, while NTA demonstrated a reproducible and heterogeneous particle population. Functional assays demonstrated that\n                    <jats:italic>P. nicotianae<\/jats:italic>\n                    EVs elicit immune-associated responses in\n                    <jats:italic>Nicotiana benthamiana<\/jats:italic>\n                    , including cell death, ROS accumulation, and callose deposition. In addition, the EVs exhibited antimicrobial activity against five soil-associated bacterial isolates. Proteomic analysis identified 2,324 proteins, revealing a diverse EV cargo enriched in metabolic enzymes, protein-modifying enzymes, and transporters, along with virulence-associated proteins. Comparative analysis across selected filamentous pathogens identified shared EV-associated protein categories, including ATP synthase subunits, elongation factors, and heat shock protein 70 (HSP70), present across all examined species. Notably, approximately 78% of EV-associated proteins from nine filamentous pathogens contained putative KFERQ-like motifs. Collectively, these findings show that\n                    <jats:italic>P. nicotianae<\/jats:italic>\n                    secretes structurally heterogeneous EV-like particles with biological activity in plant and bacterial assays, and suggest potential roles in host interaction, microbial competition, and putative conserved features that may be involved in cargo loading.\n                  <\/jats:p>","DOI":"10.3389\/fpls.2026.1899248","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T05:30:05Z","timestamp":1789104605000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Functional and proteomic characterization of Phytophthora nicotianae extracellular vesicles and putative cargo-sorting signals in filamentous pathogens"],"prefix":"10.3389","volume":"17","author":[{"given":"Celiwe I.","family":"Nxumalo","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Genetics and Microbiology; Centre for Microbial Ecology and Genomics, University of Pretoria","place":["Pretoria, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brenda C.","family":"Salasini","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Genetics and Microbiology; Centre for Microbial Ecology and Genomics, University of Pretoria","place":["Pretoria, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silindile","family":"Maphosa","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Genetics and Microbiology; Centre for Microbial Ecology and Genomics, University of Pretoria","place":["Pretoria, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucy N.","family":"Moleleki","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Genetics and Microbiology; Centre for Microbial Ecology and Genomics, University of Pretoria","place":["Pretoria, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3390\/proteomes7020023","article-title":"Different types of cellular stress affect the proteome composition of small extracellular vesicles: A mini review","volume":"7","author":"Abramowicz","year":"2019","journal-title":"Proteomes"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2019.01610","article-title":"Extracellular vesicles from the cotton pathogen Fusarium oxysporum f. sp. vasinfectum induce a phytotoxic response in plants","volume":"10","author":"Bleackley","year":"2020","journal-title":"Front. 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