title: RNAseq of barley leaf and roots exposed to control and drought stress conditions
design: RNAseq of barley leaf and roots exposed to control and drought stress conditions
library_construction_protocol: Leaf and root tissues were collected in three replicates, each consisting of three fragments of independent leaves/roots.  The plant material used in this study was the spring barley (Hordeum vulgare (L.)) cultivar 'Sebastian'. 'Sebastian' is a two-rowed cultivar and exhibits quite a high level of drought tolerance. Drought stress was applied using a previously described method (Collin et al., 2020). Briefly, we used boxes filled with a soil mixture of sandy loam and sand (7:2) with specific physicochemical properties and a nutrient medium. Plants could easily access water at a volumetric water content (vwc) of 14% in the soil, while the water deficit was 1.5%. At 14% vwc, leaf water content was approximately 100%. Soil moisture was checked daily using a time-domain reflectometer (TDR) EasyTest. For the first 10 d after sowing, plants were grown in a greenhouse with optimal water (14% vwc), 20/18°C day/night temperature, 16/8 h photoperiod, and 420 µmol m−2 s−1 PAR from fluorescent lamps. Irrigation was then withheld until the soil moisture reached 3% on the 15th day (15DAP, Days After Planting), marking the start of the drought. Plants were then moved to a chamber with a 25°C/20°C day/night regime, 16/8 h photoperiod, and 420 µEm−2s−1 light for 10 days (till 25 DAP), indicating prolonged drought at 1.5% vwc. The control plants had optimal water availability and were grown simultaneously. Drought stress was applied using a previously described method (Collin et al., 2020). Briefly, we used boxes filled with a soil mixture of sandy loam and sand (7:2) with specific physicochemical properties and a nutrient medium. Plants could easily access water at a volumetric water content (vwc) of 14% in the soil, while the water deficit was 1.5%. At 14% vwc, leaf water content was approximately 100%. Soil moisture was checked daily using a time-domain reflectometer (TDR) EasyTest. For the first 10 d after sowing, plants were grown in a greenhouse with optimal water (14% vwc), 20/18°C day/night temperature, 16/8 h photoperiod, and 420 µmol m−2 s−1 PAR from fluorescent lamps. Irrigation was then withheld until the soil moisture reached 3% on the 15th day (15DAP, Days After Planting), marking the start of the drought. Plants were then moved to a chamber with a 25°C/20°C day/night regime, 16/8 h photoperiod, and 420 µEm−2s−1 light for 10 days (till 25 DAP), indicating prolonged drought at 1.5% vwc. The control plants had optimal water availability and were grown simultaneously.  RNA was extracted using a mirVana RNA isolation kit (Thermo Fisher Scientific) according to the manufacturer's instructions. True Seq libraries were constructed for each sample. PE 150