Total RNAs were extracted from at least 5 plants of each genotype and each treatment using Qiagen Plant RNeasy extraction kit (Qiagen) and RNase free DNase I (Qiagen, Germany). Three different biological replicates were prepared. The integrity and concentration of RNA was determined using Bioanalyzer (RIN>6) and Qubit   RNA BR assay kit (Life Technologies, USA). To construct the sequencing library, 1.5   g of total RNA of each sample was used as input for the TruSeq RNA sample prep kit v2 (Illumina, USA).

Illumina HiSeq 2000 paired end sequencing; DaCBF4 overexpressor, cold treatment for 6 day, biological replicate #2

Oryza sativa

We identified the abiotic stress-responsive DaCBF4 gene in D. antarctica and generated DaCBF4-overexpressing transgenic rice plants to explore its in vivo functions. Surprisingly, Ubi:DaCBF4 plants exhibited highly increased tolerance to cold stress in terms of survival. Due to the role of DaCBF4 in transcriptional regulation, we performed a transcriptome analysis to examine the effects of cold stress on gene expression.