Title : Heterozygous rice versus Arabidopsis thaliana protein protein interactome profiling by TDOP seq
Abstract:
Protein-protein interactions (PPIs) play a crucial role in understanding molecular mechanisms in plants such as rice (Oryza sativa) and Arabidopsis thaliana. However, traditional methods for PPI network analysis often face limitations in accuracy and coverage. Here, we present an integrated approach combining Touch-down overlapping PCR with HiFi long-read sequencing (TDOP-seq) to enhance the detection and validation of PPIs. This strategy improves the assembly of complex DNA constructs, reduces amplification errors, and enables high-fidelity sequencing of interaction domains. Applying this method to rice and Arabidopsis interactome studies, we achieved comprehensive PPI mapping with enhanced resolution and reliability. We identified novel rice sodium tolerance gene OsST204, bacterial blight resistance genes OsDIR19 and OsDIR22, as well as blast resistance gene OsTPP1 by screening their protein-protein interactions with functionally characterized Arabidopsis proteins. Our results demonstrate that this synergistic technique offers a robust platform for large-scale plant interactomics, facilitating deeper insights into signaling pathways and functional genomics in crops and model plants.
