Protein kinase C is essential for viability of the rice blast fungus Magnaporthe oryzae

Abstract

SummaryProtein kinase C constitutes a family of serine–threonine kinases found in all eukaryotes and implicated in a wide range of cellular functions, including regulation of cell growth, cellular differentiation and immunity. Here, we present three independent lines of evidence which indicate that protein kinase C is essential for viability of M agnaporthe oryzae. First, all attempts to generate a target deletion of PKC 1, the single copy protein kinase C‐encoding gene, proved unsuccessful. Secondly, conditional gene silencing of PKC 1 by RNA interference led to severely reduced growth of the fungus, which was reversed by targeted deletion of the Dicer2‐encoding gene, MDL 2. Finally, selective kinase inhibition of protein kinase C by targeted allelic replacement with an analogue‐sensitive PKC 1AS allele led to specific loss of fungal viability in the presence of the PP1 inhibitor. Global transcriptional profiling following selective PKC inhibition identified significant changes in gene expression associated with cell wall re‐modelling, autophagy, signal transduction and secondary metabolism. When considered together, these results suggest protein kinase C is essential for growth and development of M . oryzae with extensive downstream targets in addition to the cell integrity pathway. Targeting protein kinase C signalling may therefore prove an effective means of controlling rice blast disease.

MoSnt2-dependent deacetylation of histone H3 mediates MoTor-dependent autophagy and plant infection by the rice blast fungus Magnaporthe oryzae

“…MoSnt2 may therefore be necessary for pathogenicity through its regulatory role in both autophagy and cell wall integrity. Signaling pathways other than the MAPK Mps1, Pkc/protein kinase C and Ca 2+ have already been shown to determine cell wall integrity and plant penetration by M. oryzae [44,47,48]. We observed that MoSnt2 is transcriptionally regulated by MoTor and mediates MoTor-dependent cellular processes.…”

“…We attempted to delete MoTOR, but did not recover any ΔMotor deletion mutants, suggesting that MoTOR is likely an essential gene. We therefore employed a conditional RNA interference (RNAi) strategy [44] to knock down expression of MoTOR. In 4 independent MoTOR RNAi transformants (pSilent1-TorFRB.7, pSilent1-TorFRB.28, pSilent1-TorFAT.39 and pSilent1-TorFAT.54),…”

Using Network Extracted Ontologies to Identify Novel Genes with Roles in Appressorium Development in the Rice Blast Fungus Magnaporthe oryzae

“…Annotations for M. oryzae were downloaded from the Ensembl Genomes Database [ 30 ]. RNA-Seq data were downloaded from the Gene Expression Omnibus (GEO) [ 31 ] from three published studies including (i) transcript profiling during appressorium formation (GSE30069) [ 19 ]; (ii) transcriptional response to hypoxia in M. oryzae (GSE51597) [ 18 ]; and (iii) a viability study with inhibited PKC1 (GSE70308) [ 32 ]. In total, the expression set contained 46 RNA-Seq samples.…”

Development of an RNA interference (RNAi) gene knockdown protocol in the anaerobic gut fungusPecoramyces ruminantiumstrain C1A

“…On the other hand, the introduction of foreign siRNA could be utilized for targeted, sequence-specific, gene knockdown in fungi ( Quoc & Nakayashiki, 2015 ; Chang, Zhang & Liu, 2012 ; Romano & Macino, 1992 ). Indeed, demonstration of the feasibility of RNAi approaches for targeted gene silencing has been shown in Ascomycota ( Romano & Macino, 1992 ; Abdel-Hadi et al, 2011 ; Barnes, Alcocer & Archer, 2008 ; Eslami et al, 2014 ; Jöchl et al, 2009 ; Kalleda, Naorem & Manchikatla, 2013 ; Li et al, 2012 ; Moazeni et al, 2012 ; Moazeni et al, 2014 ; Mousavi et al, 2015 ; Penn et al, 2015 ; Prakash, Manjrekar & Chattoo, 2016 ), Basidiomycota ( Caribé dos Santos et al, 2009 ; Matityahu et al, 2008 ; Nakade et al, 2011 ; Namekawa et al, 2005 ; Skowyra & Doering, 2012 ), and Mucoromycota ( Gheinani et al, 2011 ; Nicolas et al, 2008 ); and RNAi-based protocols were used to infer the putative roles of several genes or simply as a proof of principle.…”