RNA Sequencing analysis of Wild Type and HID1 mutant Transcriptomes

Seedling photomorphogenesis is a sophisticated developmental process that is controlled by both the transcriptional and posttranscriptional regulation of gene expression. Here, we identify an Arabidopsis noncoding RNA, designated HIDDEN TREASURE 1 (HID1), as a new factor promoting photomorphogenesisin continuous red light (cR). We show that HID1 acts through PHYTOCHROME-INTERACTING FACTOR 3 (PIF3), a bHLH transcription factor known to be a key repressor of photomorphogenesis. Overall design: Total RNA was isolated from 5-day-old cR-grown WT and hid1(hidden treasure 1)seedlings using RNeasy Plant Mini Kits (Qiagen). Three biological replicates for each sample were subjected to RNA-seq. Poly (A) RNA from 1 µg total RNA was used to generate the cDNA library according to the TruSeq RNA Sample Prep Kit protocol. The library was then sequenced using a HiSeq 2000 system (Illumina). RNA-seq reads were mapped to TAIR10 using Tophat (version 2.0.9) (1). Differentially expressed genes were defined as those that differed by >2-fold with an adjusted P value =0.05 that was identified using Cuffdiff software (version 2.0.0)