Rice blast is a pervasive and devastating disease that threatens rice production across the world. In spite of its importance to global food security, however, the underlying biology of plant infection by the blast fungus Magnaporthe oryzae remains poorly understood. In particular, it is unclear how the fungus elaborates a specialised infection cell, the appressorium, in response to surface signals from the rice leaf. Here, we report the identification of a network of temporally co-regulated transcription factors that act downstream of the Pmk1 mitogen-activated protein kinase pathway to regulate gene expression during appressorium-mediated plant infection. We show that this tiered regulatory mechanism involves Pmk1-dependent phosphorylation of the Hox7 homeobox transcription factor which regulates genes associated with induction of major physiological changes required for appressorium development, including cell cycle control, autophagic cell death, turgor generation and melanin biosynthesis, as well as controlling a further set of virulence-associated transcription factor-encoding genes. Pmk1-dependent phosphorylation of Mst12 then regulates gene functions involved in septin-dependent cytoskeletal re-organisation, polarised exocytosis and effector gene expression necessary for plant tissue invasion.

Parallel reaction monitoring assays for the temporal quantification of phosphorylation of specific residues of Hox7 in wildtype, delta-pmk1 mutant and analogue-sensitive Pmk1 variant.

Phospho-peptide were enriched by Phtalic acid-modified TiO2 enrichment from digested total protein extracts obtained from spore and appressorium samples. Time course measurements between 0 and 4hr post germination. full details provided in associated manuscript