Total RNAs were isolated from 5-DAG seedlings. More than 100 mg seedlings were collected with three biological replicates. In this assay, total RNA was isolated using the Trizol Reagent (Invitrogen Life Technologies), then concentrated. The quality and integrity were determined using a NanoDrop spectrophotometer (Thermo Scientific). Next, mRNA with poly A structure in total RNA was enriched by Oligo(dT) magnetic beads, and ion interruption was used to break the RNA into fragments of about 300 bp in length. Products were purified (AMPure XP system) and quantified using the Agilent high sensitivity DNA assay on a Bioanalyzer 2100. The first strand of cDNA was synthesized using RNA as template with 6-base random primer and reverse transcriptase, and the second strand of cDNA was synthesized using the first strand of cDNA as template. After RNA extraction, purification and library construction, the libraries were subjected to Paired-End (PE) sequencing using Next-Generation Sequencing (NGS) based on the Illumina sequencing platform. The quality-filtered reads were mapped to the Arabidopsis Information Resource (TAIR10) version of the Arabidopsis genome by Hisat2 v2.0.5 open-source software, which is freely available at http://daehwankimlab.github.io/hisat2/. We used HTSeq (0.9.1) statistics to compare the Read Count values on each gene as the original expression of the gene, and then used FPKM to standardize the expression.