Screening and validation of interaction targets for wheat E3 ubiquitin ligase TaRING1

[Objective] The study aims to explore the role of TaRING1 in the interaction between wheat and stripe rust, analyze its mechanism and provide theoretical basis for the green prevention and control of wheat stripe rust. [Methods] The interaction target proteins of TaRING1 were screened by yeast two-hybrid assays and verified by bimolecular fluorescence complementation (BiFC) and luciferase complementation (LCA) assays. The ubiquitylation function of TaRING1 was verified by ubiquitination analysis. The subcellular localization of target TaRIP92 was observed by transient expression in Nicotiana benthamiana and wheat protoplasts. [Results] Using TaRING1 as bait, the target protein TaRIP92 was screened by yeast two-hybrid, and the interaction between TaRING1 and TaRIP92 was verified by LCA and BiFC. In vitro ubiquitination assays proved that TaRING1 ubiquitinated TaRIP92. Transient expression in N . benthamiana and wheat protoplasts showed that TaRIP92 was localized in mitochondria. [Conclusion] TaRING1 interacts with and ubiquitinates the mitochondrial protein TaRIP92.