Fig. 2: Distinct electrostatic profiles of PgAFP and AfAFP and the inhibitory effects of AFPs against A. flavus conidia production in corn and peanut seeds.

A Illustration of the electrostatic surface potentials of PgAFP and AfAFP, respectively, with their orientations aligned for direct comparisons. Negatively charged regions are depicted in red, positively charged regions in blue, and neutral regions in white. The graphical representations were visualized using the PyMol APBS electrostatics plugin. Both proteins displayed predominantly positive charges on their front surfaces. However, on the back surface, PgAFP exhibited multiple negatively charged regions involving the N-terminal residues Gly8, Glu9, Cys10, Thr16, and Thr18, and the C-terminal residues Asp41, His44, Glu46, Thr58, Pro59, and Val60. In contrast, AfAFP only featured a negatively charged region known as the “oxygen hole,” comprising residues Ala35, Asn36, Arg38, Thr40, Cys57, and Lys58. The molecular weights of PgAFP and AfAFP were determined using mass spectrometry, and their isoelectric points were calculated using SnapGene software (version 6.0). The electrostatic surfaces were computed using Adaptive Poisson–Boltzmann Solver (APBS) software (version 3.4.1)⁷². B, C Inhibitory effect of heat-treated rAFPs on conidia production by A. flavus in corn and peanut seeds. (D, E) Statistical analysis of conidia production by A. flavus in (B, C). Data were presented as mean ± SEM (n = 3). Statistical significance was determined by one-way ANOVA and indicated by ** for P < 0.01. All experiments were performed in triplicate. Source data are provided as a Source Data file.