title: Analysis of extracellular vesicles produced in the biofilm by the dimorphic yeast Pichia fermentans.
abstract: The yeast Pichia fermentans DISAABA 726 strain (P. fermentans) is a dimorphic yeast that under different environmental conditions may switch from a yeast-like to pseudohyphal morphology. We hypothesize that exosomes-like vesicles (EV) could mediate this rapid modification. EV are membrane-derived vesicles carrying lipids, proteins, mRNAs and microRNAs and have been recognized as important mediators of intercellular communication. Although it has been assumed for a long time that fungi release EV, knowledge of their functions is still limited. In this work we analyze P. fermentans EV production during growth in two different media containing urea (YCU) or methionine (YCM) where yeast-like or pseudohyphal morphology are produced. We developed a procedure to extract EV from the neighboring biofilm which is faster and more efficient as compared to the widely used ultracentrifugation method. Differences in morphology and RNA content of EV suggest that they might have an active role during dimorphic transition as response to the growth conditions. Our findings are coherent with a general state of hypoxic stress of the pseudohyphal cells.
authors.affiliation: Institute of Clinical Physiology, UOS CNR, Pisa, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Leone
authors.affiliation: Department of Life Sciences, Siena, Italy.|Institute of Biology and Biotechnology CNR, Pisa, Italy.
authors.forename: Lorenza
authors.initials: L
authors.lastname: Bellani
authors.affiliation: Department of Life Sciences, Siena, Italy.
authors.forename: Simonetta
authors.initials: S
authors.lastname: Muccifora
authors.affiliation: Institute of Biology and Biotechnology CNR, Pisa, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Giorgetti
authors.affiliation: Neuroscience Department, Azienda Ospedaliero-Universitaria, Pisa, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Bongioanni
authors.affiliation: Institute of Biophysics, UOS, Pisa, Italy.
authors.forename: Marcella
authors.initials: M
authors.lastname: Simili
authors.affiliation: Plant Protection Institute CNR, Florence, Italy.
authors.forename: Biancaelena
authors.initials: B
authors.lastname: Maserti
authors.affiliation: Institute of Clinical Physiology, UOS CNR, Pisa, Italy.
authors.forename: Renata
authors.identifier: 0000-0002-7504-0379
authors.initials: R
authors.lastname: Del Carratore
mesh_terms: D018441:Biofilms; D003470:Culture Media; D000067128:Extracellular Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D005656:Fungal Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000378:metabolism; D050296:Microbial Viability; D010843:Pichia / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D012331:RNA, Fungal / Q000378:metabolism
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D035683:MicroRNAs; D012331:RNA, Fungal
keywords: Pichia fermentans; cell communication; dimorphic transition; extracellular vesicles