Small nucleolar RNA HIDDEN TREASURE 2 decrease drought tolerance through multiple pathways in Arabidopsis thaliana
To understand the role of hid2 in regulating drought tolerance, we performed the comparative transcriptome profiling WT and hid2 mutant seedlings. Overall design: 12-day-old WT and hid2 mutant seedlings were transferred from 1/2 Murashige and Skoog (MS) plates to soil and grown for 12 additional day. The rosette leaves of this 24-day-old seedlings were harvested after dehydration for 0, 2, and 4 h, and then frozen in liquid nitrogen immediately. Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA,USA) according to the manufacturer's protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and checked using RNase free agarose gel electrophoresis. After total RNA was extracted, eukaryotic mRNA was enriched by Oligo (dT) beads. Then the enriched mRNA was fragmented into short fragments using fragmentation buffer and reversly transcribed into cDNA by using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB#7530,New England Biolabs, Ipswich, MA, USA). The purified double-stranded cDNA fragments were end repaired. A base added, and ligated to Illumina sequencing adapters. The ligation reaction was purified with the AMPure XP Beads (1.0X).And polymerase chain reaction (PCR) amplified. The resulting cDNA library was sequenced using Illumina Novaseq6000.