A Protoplast‐Based Platform for Rapid and Efficient Gene Function Analysis in Non‐Heading Chinese Cabbage

Summary statement Functional genomics in non‐heading Chinese cabbage has been limited by inefficient stable transformation systems. Here, we present a versatile protoplast‐based platform that integrates transient gene expression, protein localization and interaction assays, CRISPR construct pre‐evaluation, transcriptome profiling, and transient CUT&Tag chromatin analysis. Using the cold‐responsive transcription factor BcDREB1C as a case study, this framework enables rapid identification of genome‐wide direct regulatory targets and dissection of cold‐responsive transcriptional networks. This application‐oriented and scalable strategy provides an effective route for accelerating gene function analysis and molecular breeding in NHCC and other recalcitrant Brassica crops.