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Photophthora sojae extracellular vesicles proteins

ABSTRACT: Extracellular vesicles (EVs) have been extensively studied in animal cells, and play an important role in cell-to-cell communications. Emerging evidence shows that EVs also act as important agents in plant-microbe interactions. However, the mechanisms by which EVs mediate cross-kingdom interactions between plants and microbial pathogens remain largely elusive. Here, proteomic analyses of soybean root rot pathogen Phytophthora sojae EVs identified tetraspanin family proteins, PsTET1 and PsTET3, that can be recognized by Nicotiana benthamiana to trigger plant immune responses. PsTET1 and PsTET3 were redundantly required for the full virulence of P. sojae. Further analyses revealed that the large extracellular loop (EC2) of PsTET3 is the key region recognized by N. benthamiana and also by Glycine max, and that recognition depends on the plant receptor-like kinase BAK1. TET proteins from oomycete and fungal plant pathogens could be recognized by N. benthamiana and induce immune responses. However, plant-derived TET proteins failed to do so, due to the divergent sequences of the final 16 amino acids of EC2, which ultimately makes plants distinguish self and non-self EVs, triggering active defenses against pathogenic eukaryotes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Phytophthora Sojae Strain P6497

SUBMITTER: Jinyi Zhu

LAB HEAD: Yuanchao Wang

PROVIDER: PXD040458 | Pride | 2023-08-01

REPOSITORIES: Pride

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Project description:Extracellular vesicles (EVs) are membranous particles released by different organisms. EVs carry several kinds of macromolecules implicated in cell communication. EVs have become an important topic in the study of pathogenic fungi, due to their relation with the fungal-host interactions. One of the essential research areas in this field is the characterization protein profile of EV cargo, because of plant fungal pathogens rely heavily on secreted proteins to invade their hosts. However, little is known about EVs of Botrytis cinerea, which is one of the most devastating phytopathogenic fungi. The present study has two main objectives: the first isolation and characterization of B. cinerea EVs; and the description of their potential role during the infective process. All the experimental process was conducted in B. cinerea growing in mimimal salt medium supplemented with glucose as a constitutive stage, and deproteinized tomato cell walls (TCW) as virulence inductor. The isolation of EV was performed by differential centrifugation, filtration, ultrafiltration, and sucrose cushion ultracentrifugation. EV fractions were visualised by TEM using negative staining. Proteomic analysis of EV cargo was addressed by LC-MS/MS. The used methodology had allowed, for the first time, the correct isolation of B. cinerea EVs, and the identification of a high number of EV proteins, including potential EV markers. The isolated EVs displayed differences in the morphology under both assayed conditions. GO analysis of EV proteins showed an enrichment in cell wall metabolism and proteolysis under TCW. KEGG analysis also showed difference in EV function under both conditions, highlighting the presence of potential virulence/pathogenic factors implicated in cell wall metabolism, between others. This is the first evidence and characterization of B. cinerea EVs production, showing that EVs are essential in the infection process of B. cinerea and share crucial functions with the conventional secretion pathway.

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Photophthora sojae extracellular vesicles proteins

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