Abstract
Small, secreted proteins have been found to play crucial roles in interactions between biotrophic/hemi-biotrophic pathogens and plants. However, little is known about the roles of these proteins produced by broad host-range necrotrophic phytopathogens during infection. Here, we report that a cysteine-rich, small protein SsSSVP1 in the necrotrophic phytopathogen Sclerotinia sclerotiorum was experimentally confirmed to be a secreted protein, and the secretion of SsSSVP1 from hyphae was followed by internalization and cell-to-cell movement independent of a pathogen in host cells. SsSSVP1∆SP could induce significant plant cell death and targeted silencing of SsSSVP1 resulted in a significant reduction in virulence. Through yeast two-hybrid (Y2H), coimmunoprecipitation (co-IP) and bimolecular fluorescence complementation (BiFC) assays, we demonstrated that SsSSVP1∆SP interacted with QCR8, a subunit of the cytochrome b-c1 complex of mitochondrial respiratory chain in plants. Double site-directed mutagenesis of two cysteine residues (C38 and C44) in SsSSVP1∆SP had significant effects on its homo-dimer formation, SsSSVP1∆SP-QCR8 interaction and plant cell death induction, indicating that partial cysteine residues surely play crucial roles in maintaining the structure and function of SsSSVP1. Co-localization and BiFC assays showed that SsSSVP1∆SP might hijack QCR8 to cytoplasm before QCR8 targeting into mitochondria, thereby disturbing its subcellular localization in plant cells. Furthermore, virus induced gene silencing (VIGS) of QCR8 in tobacco caused plant abnormal development and cell death, indicating the cell death induced by SsSSVP1∆SP might be caused by the SsSSVP1∆SP-QCR8 interaction, which had disturbed the QCR8 subcellular localization and hence disabled its biological functions. These results suggest that SsSSVP1 is a potential effector which may manipulate plant energy metabolism to facilitate the infection of S. sclerotiorum. Our findings indicate novel roles of small secreted proteins in the interactions between host-non-specific necrotrophic fungi and plants, and highlight the significance to illuminate the pathogenic mechanisms of this type of interaction.
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Claims 12
Claim · model-written summary
SsSSVP1 was confirmed as secreted by Sclerotinia sclerotiorum.
- Taxon
- Host Plantst0
- Interactor
- Sclerotinia sclerotiorum
Sclerotinia sclerotiorume0pathogen - Gene
- SsSSVP1e0
- Outcome
- a secreted proteine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“protein SsSSVP1 in the necrotrophic phytopathogen Sclerotinia sclerotiorum was experimentally confirmed to be a secreted protein”
t0“Host Plants”
Claim · model-written summary
SsSSVP1 entered host cells and moved between cells independently of the pathogen.
- Taxon
- Host Plantst0
- Organ / stage
- host cellse0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- SsSSVP1e0
- Outcome
- internalization and cell-to-cell movemente0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“secretion of SsSSVP1 from hyphae was followed by internalization and cell-to-cell movement independent of a pathogen in host cells”
t0“Host Plants”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
SsSSVP1∆SP significantly induced plant cell death.
- Taxon
- plante0
- Treatment
- SsSSVP1∆SPe0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- SsSSVP1∆SPe0
- Outcome
- plant cell deathe0
- Direction
- inducee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“SsSSVP1∆SP could induce significant plant cell death”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
Silencing SsSSVP1 significantly reduced virulence.
- Taxon
- Host Plantst0
- Treatment
- targeted silencing of SsSSVP1e0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- SsSSVP1e0
- Outcome
- virulencee0
- Direction
- reductione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“targeted silencing of SsSSVP1 resulted in a significant reduction in virulence”
t0“Host Plants”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
SsSSVP1∆SP interacted with plant QCR8.
- Taxon
- plantsViridiplantaee0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- QCR8e0
- Outcome
- SsSSVP1∆SP interacted with QCR8e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“SsSSVP1∆SP interacted with QCR8, a subunit of the cytochrome b-c1 complex of mitochondrial respiratory chain in plants”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
Mutating C38 and C44 in SsSSVP1∆SP significantly affected its homodimer formation.
- Taxon
- Host Plantst0
- Treatment
- Double site-directed mutagenesis of two cysteine residues (C38 and C44)e0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- SsSSVP1∆SPe0
- Outcome
- homo-dimer formatione0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Double site-directed mutagenesis of two cysteine residues (C38 and C44) in SsSSVP1∆SP had significant effects on its homo-dimer formation”
t0“Host Plants”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
Mutating C38 and C44 significantly affected SsSSVP1∆SP interaction with QCR8.
- Taxon
- Host Plantst0
- Treatment
- Double site-directed mutagenesis of two cysteine residues (C38 and C44)e0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- SsSSVP1∆SP-QCR8e0
- Outcome
- SsSSVP1∆SP-QCR8 interactione0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Double site-directed mutagenesis of two cysteine residues (C38 and C44) in SsSSVP1∆SP had significant effects on its homo-dimer formation, SsSSVP1∆SP-QCR8 interaction and plant cell death induction”
t0“Host Plants”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
Mutating C38 and C44 significantly affected plant cell death induction.
- Taxon
- plante0
- Treatment
- Double site-directed mutagenesis of two cysteine residues (C38 and C44)e0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- SsSSVP1∆SPe0
- Outcome
- plant cell death inductione0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Double site-directed mutagenesis of two cysteine residues (C38 and C44) in SsSSVP1∆SP had significant effects on its homo-dimer formation, SsSSVP1∆SP-QCR8 interaction and plant cell death induction”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
SsSSVP1∆SP may redirect QCR8 to the cytoplasm before mitochondrial targeting.
- Taxon
- plante0
- Interactor
- Sclerotinia sclerotiorumSclerotinia sclerotiorume1pathogen
- Gene
- QCR8e0
- Outcome
- subcellular localization in plant cellse0
- Qualifier
- might hijack QCR8 to cytoplasm before QCR8 targeting into mitochondriae0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“SsSSVP1∆SP might hijack QCR8 to cytoplasm before QCR8 targeting into mitochondria, thereby disturbing its subcellular localization in plant cells”
e1“Sclerotinia sclerotiorum”
Claim · model-written summary
QCR8 silencing in tobacco caused abnormal plant development.
- Taxon
- tobacco
Nicotiana tabacume0 - Treatment
- virus induced gene silencing (VIGS) of QCR8e0
- Gene
- QCR8e0
- Outcome
- plant abnormal developmente0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“virus induced gene silencing (VIGS) of QCR8 in tobacco caused plant abnormal development and cell death”
Claim · model-written summary
QCR8 silencing in tobacco caused cell death.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- virus induced gene silencing (VIGS) of QCR8e0
- Gene
- QCR8e0
- Outcome
- cell deathe0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“virus induced gene silencing (VIGS) of QCR8 in tobacco caused plant abnormal development and cell death”
Claim · model-written summary
The authors propose that SsSSVP1 acts as an effector to manipulate plant energy metabolism and aid infection.
- Taxon
- plante0
- Interactor
- S. sclerotiorumSclerotinia sclerotiorume0pathogen
- Gene
- SsSSVP1e0
- Outcome
- plant energy metabolisme0
- Qualifier
- potential effectore0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“SsSSVP1 is a potential effector which may manipulate plant energy metabolism to facilitate the infection of S. sclerotiorum”
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