The hordatines, antifungal substances previously isolated from barley seedlings grown in the dark, are also produced under conditions which approximate those in the field. The compounds, and some of their derivatives, inhibit the spore germination of a number of fungi in concentrations as low as 10−5 M. Inhibition is reversed by the addition of some divalent cations; this observation supports the identity of the hordatines with the natural defense agents of barley which have been demonstrated in earlier work.