Tissue-specific mRNA profiling of the Brassica napus-Sclerotinia sclerotiorum interaction uncovers novel regulators of plant immunity

Abstract

White mold in Brassica napus (canola) is caused by the fungal pathogen Sclerotinia sclerotiorum and is responsible for significant losses in crop yield across the globe. With advances in high-throughput transcriptomics, our understanding of the B. napus defense response to S. sclerotiorum is becoming clearer; however, the response of individual tissue layers directly at the site of infection has yet to be explored. Using laser microdissection coupled with RNA sequencing, we profiled the epidermis, mesophyll and vascular leaf tissue layers in response to S. sclerotiorum. This strategy increases the number of genes detected compared to whole-leaf assessments and provides unprecedented information on tissue-specific gene expression networks in response to pathogen attack. Our findings provide novel insight into the conserved and specific roles of ontogenetically distinct leaf tissue layers in response to infection. Using bioinformatics tools, we identified several defense genes that might coordinate plant immunity responses shared across different tissue layers within the leaf. These genes were functionally characterized by challenging T-DNA insertion lines of Arabidopsis with necrotrophic, hemi-biotrophic, and biotrophic pathogens, ultimately converging on the PR5-like RECEPTOR KINASE (PRK5). Together, these data provide insight on the complexity of the B. napus defense response directly at the site of infection.