RNA-seq analysis during adventitious root formation in cuttings of PuHox52 overexpression and wild type Populus ussuriensis.

RNA-seq was performed to examine the differential expressed transcriptomes with five-point experiment (6, 12, 24, 48 and 96 h) at the stem bases of cuttings in PuHox52 overexpression line compare to wild type Populus ussuriensis. Overall design: For the RNA-seq experiments of PuHox52-mediated adventitious root formation, two week-old WT and 35S::PuHox52 in vitro plants were grown on the 1/2 MS medium at different rooting stages (6, 12, 24, 48 and 96 h) and harvested the basal ends of stem cuttings (0.5 cm). Samples were collected from two biological replicates (40 cuttings each). Total RNA isolation, library construction and sequencing were performed by Biomarker Technologies (Beijing, China). A total amount of 1 µg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (#E7530L, NEB, USA) following manufacturer's recommendations. The library preparations were sequenced on an Illumina Hiseq 2500 platform and paired-end reads were generated. About 4 Gb of high-quality 125-bp paired-end reads were generated from each library.