Illumina HiSeq 2500 sequencing; RNA-seq of leaf tissue of barley cultivar Bowman and three derived introgression lines  carrying mutations in HvELF3 (BW290), HvLUX1 (BW284) and EARLY MATURITY 7 (EAM7) (BW287) loci grown under diel (12h light / 12h dark) and constant light conditions
RNA-seq of leaf tissue of barley cultivar Bowman and three derived introgression lines  carrying mutations in HvELF3 (BW290), HvLUX1 (BW284) and EARLY MATURITY 7 (EAM7) (BW287) loci grown under diel (12h light / 12h dark) and constant light conditions
After three weeks of growth, the second expanded leaf after the cotyledon leaf was harvested for two replicate plants per genotype and time point every 4h for 24h starting with lights off. For Bowman, an extra time-point at 14-h was taken. After completion of a 12h dark/12h light diel cycle, the growth chamber was switched to constant light and 20°C and sampling continued for 36 h more every 4 hours starting from the first subjective dusk. Four spring barley genotypes were used, the wild-type spring barley Bowman (BW) and three derived introgression lines with mutations in HvELF3 (BW290), HvLUX1 (BW284) and EARLY MATURITY 7 (EAM7) (BW287). The lines were sown in soil (Einheitserde) in 96-well format. Seeds were maintained at 4°C for three days, followed by germination in 12 h light/12 h dark photoperiods at 20°C with a photon flux density 300µmol m-2 s-1 during the day and 18°C during the night and grown for three weeks. Total RNA was extracted from ground leaf tissue using a hybrid protocol of TRIZOL (Invitrogen) and purification columns from an RNeasy RNA extraction kit (Qiagen). Extracted total RNA was DNAse treated (Ambion). The concentration and integrity of the extracted RNA were determined on a BioAnalyzer (Agilent) before library preparation. The library preparation was carried out following the TruSeq protocol.
Experimental Factor: genotype
HvLUX1 mutant
Experimental Factor: time
68
Experimental Factor: environmental history
continuous light regimen
Experimental Factor: temperature
20