fill N. benthamiana isa.study

studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/protocols/BlumeriaHordei.md

 ## Blumeria hordei
-*Blumeria hordei* isolate K174 was maintained on intact plants of barley cv. Golden Promise grown in soil at 19 °C, 60% relative humidity and a photoperiod of 14 h with a light intensity of 100 μmol ·m-2 s-1. Control and *GBP* knock-out lines were grown in soil in a growth chamber (poly klima) at 19 °C, 60% relative humidity and a photoperiod of 14 h with a light intensity of 100 μmol ·m−2·s−1. Primary leaves of seven-day-old plants were cut and placed with the adaxial side down on 1% plant agar plates before gravity inoculation with *B. hordei* isolate K1 at a conidial density of about 20 conidia per mm. Leaves were harvested at 48 h and cleared in 70% ethanol before staining of the fungal structures with Coomassie brilliant blue solution (0.1% [w/v] Coomassie brilliant blue R-250 in 50% ethanol and 10% acetic acid) for 10-15 s. Bright field microscopy was used to assess secondary hyphae formation in 50 germinated conidia spores in the tip area and 50 germinated conidia in the middle of the leaf. At least six independent leaves were examined at 48 h after infection for fungal penetration success.
+*Blumeria hordei* isolate K1 (Hinze et al., 1991) was maintained on intact plants of barley cv. Golden Promise grown in soil at 19 °C, 60% relative humidity and a photoperiod of 14 h with a light intensity of 100 μmol ·m-2 s-1. Control and *GBP* knock-out lines were grown in soil in a growth chamber (poly klima) at 19 °C, 60% relative humidity and a photoperiod of 14 h with a light intensity of 100 μmol ·m−2·s−1. Primary leaves of seven-day-old plants were cut and placed with the adaxial side down on 1% plant agar plates before gravity inoculation with *B. hordei* isolate K1 at a conidial density of about 20 conidia per mm. Leaves were harvested at 48 h and cleared in 70% ethanol before staining of the fungal structures with Coomassie brilliant blue solution (0.1% [w/v] Coomassie brilliant blue R-250 in 50% ethanol and 10% acetic acid) for 10-15 s. Bright field microscopy was used to assess secondary hyphae formation in 50 germinated conidia spores in the tip area and 50 germinated conidia in the middle of the leaf. At least six independent leaves were examined at 48 h after infection for fungal penetration success.
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studies/HordeumVulgare/protocols/PlantMaterialAndGrowthConditions.md

 ## PlantMaterialAndGrowthConditions
-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 (Gol et al., 2021) 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).
+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 (Gol et al., 2021) 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). 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).
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