The Type III Effectome of the Symbiotic Bradyrhizobium vignae Strain ORS3257
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains and Growth Conditions
2.2. Plasmid Construction and Mutagenesis
2.3. Transcriptome Analysis
2.4. Proteome Analysis
2.5. Western Blot Analysis
2.6. Symbiotic Tests
2.7. Outer Membrane Vesicles Observation by Transmission Electron Microscopy
3. Results
3.1. Transcriptome Identification of the Genes under the Control of TtsI and the Flavonoid Genistein in Bradyrhizobium vignae ORS3257
3.1.1. T3SS Machinery and Predicted T3SS Effectors Genes Regulated by TtsI
3.1.2. Identification of New Putative Effectors by Transcriptome Analysis
3.2. T3SS-Dependent Secretome in Bradyrhizobium vignae ORS3257
3.2.1. Cytosolic and Membrane Proteins Are Present in the Supernatant
3.2.2. The Expression Level of the T3SS Apparatus Modulates Protein Production and Secretion in Bradyrhizobium vignae ORS3257
3.2.3. T3SS-Dependent Secreted Proteins by Bradyrhizobium vignae ORS3257 and Identification of New Putative Effectors
3.2.4. Validation of T3SS-Effector Candidates Identified by Omics Using Western-Blotting
3.2.5. Secretion of T3SS-Needle Components
3.3. A Mutant in the BRAD3257_v2_7792 Effector Is Not Impaired in Symbiosis with Aeschynomene indica
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Busset, N.; Gully, D.; Teulet, A.; Fardoux, J.; Camuel, A.; Cornu, D.; Severac, D.; Giraud, E.; Mergaert, P. The Type III Effectome of the Symbiotic Bradyrhizobium vignae Strain ORS3257. Biomolecules 2021, 11, 1592. https://doi.org/10.3390/biom11111592
Busset N, Gully D, Teulet A, Fardoux J, Camuel A, Cornu D, Severac D, Giraud E, Mergaert P. The Type III Effectome of the Symbiotic Bradyrhizobium vignae Strain ORS3257. Biomolecules. 2021; 11(11):1592. https://doi.org/10.3390/biom11111592
Chicago/Turabian StyleBusset, Nicolas, Djamel Gully, Albin Teulet, Joël Fardoux, Alicia Camuel, David Cornu, Dany Severac, Eric Giraud, and Peter Mergaert. 2021. "The Type III Effectome of the Symbiotic Bradyrhizobium vignae Strain ORS3257" Biomolecules 11, no. 11: 1592. https://doi.org/10.3390/biom11111592