RNAseq analyses of the multiple mutant plants of CCA1/LHY and RVE4/RVE8 under cold stresses

It was reported that CCA1 and LHY positively regulated the gene expression of DREB1A and DREB1C. In addition, we revealed that RVE4 and RVE8 directly activated the cold-inducible DREB1A expression. The cca1 lhy double mutant, rve4 rve8 double mutant (rve48), and cca1 lhy rve4 rve8 quadruple mutant (quad) plants were sensitive to the freezing stress. In order to comprehensively analyze downstream genes of RVE4/RVE8 and CCA1/LHY in the cold response, we conducted RNA sequence analyses. We used 14-day-old whole seedlings of wild type (Col-0), cca1 lhy, rve48 and quad mutant plants grown on GM agar plates under the normal growth conditions at ZT2 (2 h after dawn) and the cold stress conditions treated at 4°C for 3 h and 12 h. We have three biological replicates for this analysis.