Proteomics Analysis of SsNsd1-Mediated Compound Appressoria Formation in Sclerotinia sclerotiorum

Abstract

Sclerotinia sclerotiorum (Lib.) de Bary is a devastating necrotrophic fungal pathogen attacking a broad range of agricultural crops. In this study, although the transcript accumulation of SsNsd1, a GATA-type IVb transcription factor, was much lower during the vegetative hyphae stage, its mutants completely abolished the development of compound appressoria. To further elucidate how SsNsd1 influenced the appressorium formation, we conducted proteomics-based analysis of the wild-type and ΔSsNsd1 mutant by two-dimensional electrophoresis (2-DE). A total number of 43 differentially expressed proteins (≥3-fold change) were observed. Of them, 77% were downregulated, whereas 14% were upregulated. Four protein spots fully disappeared in the mutants. Further, we evaluated these protein sequences by mass spectrometry analysis of the peptide mass and obtained functionally annotated 40 proteins, among which only 17 proteins (38%) were identified to have known functions including energy production, metabolism, protein fate, stress response, cellular organization, and cell growth and division. However, the remaining 23 proteins (56%) were characterized as hypothetical proteins among which four proteins (17%) were predicted to contain the signal peptides. In conclusion, the differentially expressed proteins identified in this study shed light on the ΔSsNsd1 mutant-mediated appressorium deficiency and can be used in future investigations to better understand the signaling mechanisms of SsNsd1 in S. sclerotiorum.

Secretome-Wide Analysis of Lysine Acetylation in Fusarium oxysporum f. sp. lycopersici Provides Novel Insights Into Infection-Related Proteins

“…The primer pairs used for real-time PCR are listed in Supplementary Data 1 . Hierarchical clustering was performed using the MeV program ( Li et al, 2014 , 2018b ). All graphs were exported by the GraphPad Prism 6 software (La Jolla, CA, United States).…”

The Formaldehyde Dehydrogenase SsFdh1 Is Regulated by and Functionally Cooperates with the GATA Transcription Factor SsNsd1 in Sclerotinia sclerotiorum

“…flavus, NsdD is essential for transcriptional control of a series of genes that are vital for the morphological development of sclerotia and the production of conidia (43). In this regard, SsNsd1could also regulate several downstream targets during compound appressorium formation (12).…”

Diversity and Function of Appressoria

“…Genomics and their interconnected disciplines have provided a fertile platform to address most of these issues and controversies related to the formation and functioning of appressoria. Genomic data and novel techniques have contributed towards a better understanding of these fungal-host interactions from appressorium formation to host invasion [28][29][30][31][32]. Advances in reverse genetic techniques have enabled cloning of diverse genes predicted to be involved in these interactions, and many are characterized via various functional genomic approaches [17,21,28,[33][34][35][36][37][38][39][40][41][42].…”