RNA-seq analysis of flg22-induced gene expression changes in wild type, asr3-1 mutant and OX9 overexpression line

The correlation coefficient (R) for the expression profile of all transcripts between WT and asr3-1, and between WT and OX9 without flg22 treatment is close to linear, suggesting that ASR3 does not affect general gene transcription. Hierarchical clustering analysis flg22-induced genes in any of the three genotypes suggested that the asr3-1 mutant displayed overall enhanced flg22 response whereas OX9 displayed overall reduced responses compared to WT plants. RNA-seq analysis was performed with 10-day-old seedlings of WT, asr3-1 and 35S:ASR3-HA transgenic line OX9 with or without 100 nM flg22 treatment for 30 min. Two independent repeats were performed for RNA-seq analysis. For each repeat, equal amount of RNA from two biological replicates was pooled for RNA-seq library construction. RNA-seq library preparation and sequencing were carried out on an Illumina HiSeq 2500 platform with 100 nt single end reads.