Abstract

Low temperature as one of the most important abiotic stresses has caused serious wheat production loss worldwide. We identified TaWRKY115 affecting cold tolerance in wheat by integration of genome-wide association study and RNA sequencing. Overexpression and CRISPR/Cas9-mediated gene-editing revealed TaWRKY115 positively modulating cold tolerance in wheat. Transcription factor TaSP1 negatively regulates the cold tolerance of wheat by binding to the promoter of TaWRKY115 in cold-sensitive haplotypes. TaWRKY115 inhibited TaMYB4 at both protein and transcription levels. BSMV-mediated silenced wheat plants and EMS mutants of TaMYB4 possessed increased wheat cold tolerance. We further revealed that TaMYB4 repressed expressions of the TaCBF family through binding to their promoters. BSMV-mediated silenced wheat plants and EMS mutants of TaCBF12d exhibited decreased cold tolerance in wheat. This study brought forth a new model TaSP1-TaWRKY115-TaMYB4, regulating wheat cold tolerance through the TaCBF pathway and provided valuable cold tolerance genes for wheat breeding programs.