HordeumVulgare.md

Hordeum vulgare

All experiments, including the generation of CRISPR/Cas9 knock-out lines, were performed with the spring barley (H. vulgare L.) cv. Golden Promise Fast, an introgression line carrying the Ppd-H1 allele that confers fast flowering.42 From here on, we use “control” to name this non-mutagenized cultivar that carries normal copies of GBP1 and GBP2. For ROS burst assays, barley seeds were surface sterilized with 6% sodium hypochlorite for 1 h and then washed extensively (5 × 30 mL sterile water). Seeds were germinated on wet filter paper at room temperature in the dark under sterile conditions for three days before transfer to sterile jars containing solid 1/10 plant nutrition medium (PNM), pH 5.7 and 0.4% Gelrite (Duchefa, Haarlem, the Netherlands).82 Seedlings were cultured for four days in a growth chamber under long-day conditions (day/night cycle of 16/8 h, 22 °C/18 °C, light intensity of 108 μmol·m−2·s−1).