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