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Commit 6f02f331 authored by Viktoria Petrova's avatar Viktoria Petrova
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add FungalMaterialGrowthConditionsAndBarleyColonizationAssays assay

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## Blumeria hordei
*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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## Rhizophagus irregularis
Barley control and mutant seeds were sterilized and germinated as described previously (Mahdi et al., 2022). Germinated seedlings were transferred to 9 × 9 cm pots containing autoclaved 1:1 silica sand:vermiculite mixture inoculated with 700 mg *R. irregularis* spore inoculum (10,000 spores·g-1 moist diatomaceous earth powder [50% water]) (Symplanta, Darmstadt, Germany). Approximately 500 mg of the inoculum was evenly mixed into the lower two-third substrate layer, further 200 mg were evenly sprinkled into a hole in the upper third of the substrate layer, into which the seedlings were transplanted. The seedlings were grown in the greenhouse and watered weekly with 30 mL of deionized water and tap water in a 1:1 ratio. Roots were harvested at 28 dpi and stored in 50% EtOH at 4 °C until staining.
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## Serendipita indica, Serendipita vermifera and Bipolaris sorokiniana
The growth conditions for barley, *S. indica* (DSM11827, Si), *S. vermifera* (MAFF305830, Sv) and *B. sorokiniana* (ND90Pr, Bs) and the preparation of fungal suspensions for plant inoculation have been described previously (Sarkar et al., 2019; Mahdi et al., 2022). Four-day-old barley seedlings were transferred to sterile jars on 1/10 PNM (pH 5.7) and inoculated with 3 mL of either sterile water as control, *Si* chlamydospores (500,000 spores·mL-1), *Sv* mycelium (1 g per 50 mL), or *Bs* conidia (5,000 spores·mL-1). Plants were grown on a day/night cycle of 16/8 h at 22/18 °C and 60% humidity under a light intensity of 108 μmol·m−2·s−1. Plant roots were harvested six days post inoculation (dpi), washed thoroughly to remove extraradical fungal hyphae, and frozen in liquid nitrogen. Four barley plants were pooled per biological replicate.
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