Multi-Omics and PTM Atlas of Wheat Responses to Dynamic Stripe Rust Infections

Abstract

Wheat stripe rust, caused by the obligate biotrophic fungus Puccinia striiformis f. sp. tritici (Pst), poses a catastrophic threat to global wheat production. While host transcriptional and translational variations have been extensively investigated, the dynamic, multi-layered regulatory networks governed by post-translational modifications (PTMs) during wheat-Pst interactions remain largely uncharacterized. To bridge this gap, we deployed a comprehensive, integrated systems-biology approach combining transcriptomic, proteomic, phosphoproteomic, ubiquitinomic, and acetylomic analyses to resolve wheat immune and metabolic dynamics during both compatible (susceptible, inoculated with Pst race CYR32) and incompatible (resistant, inoculated with Pst race CYR25) interactions. Leaf samples were systematically harvested from mock-treated, CYR25-infected, and CYR32-infected wheat plants at three critical developmental milestones: 12, 24, and 168 hours post-inoculation (hpi), spanning three independent biological replicates per treatment and timepoint. Total RNA was profiled using high-throughput sequencing on the Illumina platform. In parallel, full-proteome and targeted PTM peptides (individually enriched for phosphorylation, ubiquitination, and acetylation) were quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a high-resolution mass spectrometry platform. This multi-omics dataset captures global transcriptional reprogramming, progressive protein abundance alterations, and wide-ranging multi-PTM cascades in a highly coordinated, time-resolved manner. Collectively, this comprehensive multi-omics repository provides a foundational and monumental resource for dissecting multi-layered, PTM-mediated crop immunity and supports the systematic exploration of host-pathogen dynamics in hexaploid wheat.