RNAseq of barley leaf and roots exposed to control and drought stress conditions
RNAseq of barley leaf and roots exposed to control and drought stress conditions
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