Cx40 ph

To collect fungal spores, 10 ml of 0.1(v/v) % Tween 80^® solution was poured onto a culture plate and the surface of the fungal colony gently scraped to release the spores. The spore suspension was collected using a single channel automatic micropipette and stored at 4 °C for 3–5 days. The spore concentration was estimated using Haemocytometer (Bright-Line^®, Z359629) and a light microscope (Olympus, CX40-PH).
Spore viability was analyzed following a protocol modified from (Tupe et al. 2017 (link)). The spore suspensions were diluted into 1–3 × 10^–3 spore/ml concentration with 0.1%(v/v) Tween 80^® solution, then the diluted spore suspension was spread (100 µL) onto SDAY media. After 2–5 days of incubation the number of colonies were counted, and spore viability was estimated. For the bioassay, a spore concentration at ≥ 10⁷ conidia/ml was chosen (following Tupe et al. 2017 (link)). In addition, the work of Mubeen et al. (2022 (link)) suggests that there is no statistically significant mortality difference for fungal spore concentrations of 10⁷–10⁹ conidia/ml in S. frugiperda. We also ensured that the spore viabilities of all candidates were ≥ 85%. To generate reliable data, the spore count and viability was performed in three technical replications.

To collect fungal spores, 10 ml of 0.1(v/v) % Tween 80^® solution was poured onto a culture plate and the surface of the fungal colony gently scraped to release the spores. The spore suspension was collected using a single channel automatic micropipette and stored at 4 °C for 3–5 days. The spore concentration was estimated using Haemocytometer (Bright-Line^®, Z359629) and a light microscope (Olympus, CX40-PH).
Spore viability was analyzed following a protocol modified from (Tupe et al. 2017 (link)). The spore suspensions were diluted into 1–3 × 10^–3 spore/ml concentration with 0.1%(v/v) Tween 80^® solution, then the diluted spore suspension was spread (100 µL) onto SDAY media. After 2–5 days of incubation the number of colonies were counted, and spore viability was estimated. For the bioassay, a spore concentration at ≥ 10⁷ conidia/ml was chosen (following Tupe et al. 2017 (link)). In addition, the work of Mubeen et al. (2022 (link)) suggests that there is no statistically significant mortality difference for fungal spore concentrations of 10⁷–10⁹ conidia/ml in S. frugiperda. We also ensured that the spore viabilities of all candidates were ≥ 85%. To generate reliable data, the spore count and viability was performed in three technical replications.