Table 1_Genome-wide analysis of the EIN3/EIL family in rye and functional identification of ScEIL5 in stripe rust resistance.xlsx

IntroductionIntroduction: Rye (Secale cereale L.) is a cereal crop with superior overall stress tolerance, particularly prominent resistance to stripe rust, cold, and drought. It thus serves as an important donor of resistance genes for other cereal species. Ethylene-insensitive 3/ethylene-insensitive 3-like (EIN3/EIL) proteins are core transcription factors that transduce ethylene signals into downstream transcriptional cascades. However, their systematic characterization and defense-related functions in rye remain unexplored.
MethodsHere, we performed genome-wide identification of ScEIL genes from the rye reference genome, designated them based on chromosomal distribution, and analyzed their gene structure, collinearity, and cis-elements. We also examined the expression profiles of ScEIL genes under biotic and abiotic stresses, determined the subcellular localization of ScEIL5, and verified its function via VIGS-mediated silencing.
ResultsSeven ScEIL genes (ScEIL1–ScEIL7) were identified. Gene structure analysis revealed a highly conserved N-terminal DNA-binding domain and divergent C-terminal regions. Collinearity analysis showed significant evolutionary divergence of the EIN3/EIL family between monocotyledonous and dicotyledonous plants. Cis-element prediction and expression profiling indicated that ScEIL genes are involved in rye responses to various stress conditions and exhibit significant tissue specificity, with the lowest expression detected in seeds. ScEIL5 is localized to the nucleus and showed approximately 4.8-fold higher expression at 48 h post-inoculation with Puccinia striiformis f. sp. tritici compared with 0 h. VIGS-mediated silencing of ScEIL5 enhanced stripe rust resistance and reduced fungal biomass.
DiscussionThese results suggest that ScEIL5 acts as a negative regulator of stripe rust resistance in rye. This study not only provides valuable insights for further research on the EIN3/EIL gene family, but also offers important clues regarding the role of ScEIL genes in rye’s response to stripe rust stress.