A new peptide of melon seeds which shows sequence homology with vicilin: Partial characterization and antifungal activity

Plant resistance against pathogens involves a broad variety of proteins, which can be either constitutive or induced. Different types of antimicrobial proteins have been purified from plant seeds, including chitinases, β-1,3-glucanases, defensins, thionins, lipid transfer proteins, lectins and vicilins. The aims of this study were to isolate and characterize defense-related proteins, particularly antimicrobial peptides, present in melon (Cucumis melo L.) seeds. Seed proteins were extracted with phosphate buffer, pH 5.4 and subjected to precipitation with 90% relative ammonium sulfate saturation. The precipitate was re-dissolved in distilled water and heated at 80 °C for 15 min. A DEAE-Sepharose column equilibrated with 0.1 M Tris-HCl pH 8.0 was employed for further purification of the peptides with antifungal properties. Fractions with inhibitory activity were detected in the non-retained basic peak (P1). To test whether P1 peptides were able to permeabilize the yeast plasma membrane, we monitored their effects on glucose-stimulated acidification of the incubation medium by yeast Saccharomyces cerevisiae cells. This fraction strongly inhibited glucose-stimulated acidification of the medium by yeast cells in a dose-dependent manner, and inhibition was up to 60% for all concentrations tested. Effects of the P1 fraction upon the growth of the phytopathogenic fungus Fusarium oxysporum were also observed. Proteins in P1 fraction were also subjected to automated N-terminal amino acid sequencing. Partial N-terminal sequence of a major 8 kDa protein revealed homology with previous reported storage vicilins from different seeds.