Seven toxins (F1-F7) were purified from Tityus discrepans scorpion venom on a C18 HPLC column. The compounds were fungitoxic on Macrophomina phaseolina. The molecular masses of F1-F7 were (Da) 1061.1, 7328.8, 7288.3, 7268.5, 7104.6, 6924.6, and 6823.3, respectively. It is not known if F1 is a small peptide or some other kind of organic molecule. Compounds F2-F7 were peptides. The most potent was F7, with a minimal inhibition concentration of 0.4 μg/μL and a concentration for 50% inhibition of 0.13 μg/μL. Fungal esterase activity was abolished by F2, F3, and F5 and inhibited by 89, 60, 58, and 54% by F4, F6, F7, and F1, respectively. F1, F2, F5, and F7 induced an increase on hyphae chitin wall and septum thickness. Peptides F3-F6 induced efflux of the fluorescent dye Na-CoroNa Red complex from hyphae. Only F5 and F6 were inhibited by the prokaryote sodium channel blockers amiloride and mibefradil. Gas chromatography-mass spectrometry analysis suggested that F1, F5, F6, and F7 altered sterol biosynthesis either by inhibiting ergosterol biosynthesis or by producing ergosterol analogues. The peptides affect M. phaseolina viability by three mechanisms: decreasing esterase activity, altering Na(+) membrane permeability, and altering wall sterol biosynthesis. It seems that interfering with sterol synthesis is an important mechanism behind the effect of the fungicideal toxins. However, the antifungal effects at short times are indicative of a direct esterase inhibition, which, with the increased membrane leakiness to Na(+), makes the fungus inviable.