Table 4 A method comparison of the optimized conidia production in Pyrenophora sp. (this study) to the Lamari and Bernier 29 protocol

Vegetative Growth

Grow stock culture from infected leaf sample in V8-PDA (4–8 days)

Grow freeze dried mycelial plug in V8-PDA (5–7 days)

Conidiophore Production

Transfer and Incubate in the dark single mycelial plugs on V8-PDA (3–5 days)

Transfer 16 plugs to V8-PDA and incubate at N-UV (24 h)

Flood culture with water and flatten mycelia

Decant water and incubate in light at RT (18–24 h)

Decant water and incubate in light at RT (18–24 h)

Conidia Production

Subsequent dark incubation at 15 °C (18–24 h)

Subsequent dark incubation at 15 °C (18 h)

Spore Recovery

Flood plate with water and dislodge conidia with wire loop

Remove excess agar and liberate conidia with fine paint brush in a Tween 20 solution

Two to three additional water rinses

Spin down (5 min)

Incubate spore suspension at 5 °C (10–15 min)

Decant excess water and add Tween 20