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. 2023 Jun 5;89(6):e00705-23. doi: 10.1128/aem.00705-23

FIG 2.

FIG 2

Expression of BbVRF1 and AMP expression levels after infection. (A) PCR detection for the presence of the fragment (cassette gpdA promoter + Bbsp + VRF1 + mCherry + trpC terminator) in the genomic DNA of the wild-type (WT) strain (negative control) and transformant BbVRF1. M, marker. (B) Semi-quantitative RT-PCR analysis of VRF1 in the cDNA of WT and BbVRF1. The actin gene was used as an internal reference. (C) Western blot of the secretory venom protein VRF1 in Sabouraud dextrose broth (SDB) of WT and BbVRF1. M, marker. (D to F) qRT-PCR analysis of several AMP genes regulated by the Toll pathway in IOZCAS-Ha-I cells (H. armigera cell line) 72 h after coculture with B. bassiana strains. Mock, cells infected with 1 × PBS. WT, cells infected with WT (B. bassiana ARSEF 2860). BbVRF1, cells infected with transformant BbVRF1. Data are shown as means ± standard deviation (SD) of three biological replicates. Ordinary one-way analysis of variance (ANOVA) with Tukey’s multiple-comparison test was performed (P ≤ 0.05).