In planta colocalization and induction of gene expression in wood-degrading fungi
EMSL Project ID
48929
Abstract
Wood-degrading fungi make energy from nature’s most abundant yet most recalcitrant plant tissue.9 Their mechanisms have obvious bioenergy relevance, and the brown rot fungi we study accomplish this without removing large amounts of lignin and without exo-acting cellulases. Aided by a Special Call, Quiet Wing proposal, we have begun working at EMSL using Helium Ion Microscopy (HIM) and single-molecule fluorescence in situ hybridization (smFISH) to colocalize dynamics in planta. This is already bearing fruit, with smFISH images for a Nature Communications submission. We would like to deepen the effort by integrating Quiet Wing microscopy and transcriptomics capabilities at EMSL.We have two current DOE BER bioenergy-focused efforts probing spatial dynamics in brown rot fungi. These fungi have long been theorized to deploy oxidative radical-based ‘pretreatments’ prior to secreting cellulases into the wood cell wall,1,2,3 but mechanisms are putative, in part due to limitations studying in planta.4 Since 2010, we have used coarse (mm-scale) spatial differentiation of wood physiochemistry and mRNA via qPCR to show evidence that reactions are indeed partitioned, perhaps segregated inside wood cell walls.5 Our goal at EMSL is to test two long-standing hypotheses in planta that we are addressing using cutting-edge EMSL instrumentation and technical collaboration. Aim 1) Test the Shield hypothesis - Reaction partitioning between fungal hyphae and wood cell walls is enabled by leaving a thin tertiary wall layer (S3) intact. Aim 2) Test the Differential hypothesis - Hyphal tips express oxidative-enabled pathways, specifically Fenton reaction iron-reduction genes, ahead of cellulolytic enzyme expression.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2015-10-01
End Date
2017-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Zhang J., D. Hu, G. Orr, and J.S. Schilling. 2019. "Fluorescence In Situ mRNA Hybridization for Gene Expression Detection in a Wood Decay Fungus." International Biodeterioration & Biodegradation 143. PNNL-SA-140382. doi:10.1016/j.ibiod.2019.104731
Zhang J, GN Presley, KE Hammel, JS Ryu, J Menke, M Figueroa, D Hu, G Orr, and JS Schilling. 2016. "Localizing gene regulation reveals a staggered wood decay mechanism for the brown rot fungus Postia placenta." Proceedings of the National Academy of Sciences of the United States of America 113(39):10968-10973. doi:10.1073/pnas.1608454113