In order to determine DON content of strains, the conidia solution of PH-1 and FgMet3, FgMet14 deletion mutants (2×103 conidia/mL) were transferred into the toxin production medium GYEP and cultured at 28°C, 175rpm in the dark condition for 7 days (Bian et al., 2021 (link)). Then the DON content of strains was determined by DON ELISA Plate Kit (Weisai) (Duan et al., 2020 (link)). The DON content (μg/g) of the strain was expressed by the DON content per unit weight of hyphae.
To determine the expression of Tri5, Tri6, which related to DON synthesis, the conidia of PH-1 and FgMet3, FgMet14 deletion mutants were placed in GYEP medium at 28°C, 175rpm for 36 hours in the dark. The hyphae were collected, RNA was extracted by RNA extraction kit, and inversely converted to cDNA with HiScript II Q RT SuperMix (+ gDNA wiper) (Vazyme Biotech Co. Ltd)., then the expression of Tri5, Tri6 in the strain was determined by qPCR (Bian et al., 2021 (link)). FgGadph was used as the reference gene. The experiment was repeated three times.
To determine the expression of Tri5, Tri6, which related to DON synthesis, the conidia of PH-1 and FgMet3, FgMet14 deletion mutants were placed in GYEP medium at 28°C, 175rpm for 36 hours in the dark. The hyphae were collected, RNA was extracted by RNA extraction kit, and inversely converted to cDNA with HiScript II Q RT SuperMix (+ gDNA wiper) (Vazyme Biotech Co. Ltd)., then the expression of Tri5, Tri6 in the strain was determined by qPCR (Bian et al., 2021 (link)). FgGadph was used as the reference gene. The experiment was repeated three times.
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