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Cold stress is a major environmental factor that impairs plant growth and development, geographic distribution, and crop productivity. The C-repeat binding factor (CBF) regulatory pathway has an essential role in response to cold stress. Here, we characterized a bHLH transcription factor from  Nicotiana tabacum ,  NtbHLH123 , in response to cold stress (4°C). Overexpression of  NtbHLH123  enhanced cold tolerance in transgenic tobacco plants. Based on yeast one-hybrid, chromatin immunoprecipitation PCR, and transient expression analysis assays,  NtbHLH123  binds directly to the G-box/E-box motifs in the promoter of the  NtCBF  genes and positively regulates their expression. Furthermore,  NtbHLH123 -overexpressing plants showed lower electrolyte leakage, reduced malondialdehyde contents, H 2 O 2  and reactive oxygen species (ROS) accumulation under cold stress, which contributed to alleviating oxidative damage to the cell membrane after cold stress treatment. And  NtbHLH123  increased stress tolerance by improving the expression of a number of abiotic stress-responsive genes to mediate the ROS scavenging ability and other stress tolerance pathways. Taken together, we present a model suggesting that  NtbHLH123  is a transcriptional activator that functions as a positive regulator of cold tolerance by activating  NtCBF , ROS scavenging-related, and stress-responsive genes.

Tobacco Transcription Factor NtbHLH123 Confers Tolerance to Cold Stress by Regulating the NtCBF Pathway and Reactive Oxygen Species Homeostasis