Action of methyl-2-benzimidazolecarbamate (MBC), a breakdown product of benomyl, was studied in synchronous cultures of Ustilago maydis and Saccharomyces cerevesiae. Cells treated with MBC developed similarly to control cells until the two portions of the doublets (joined mother and daughter “cells”) were morphologically equivalent. At this point development of the treated cells ceased but control doublets separated to form two new cells. The compound did not prevent DNA synthesis during the first cell cycle of synchronized S. cerevesiae cells but did prevent completion of cell division, as it did also in synchronized U. maydis sporidia which already contained the DNA complement needed for completion of the first division. Mitosis did not go to completion in MBC-treated cells so the doublets formed contained only a single compact nucleus. The effect of MBC markedly resembles mitotic arrest caused by colchicine and isopropyl N-phenylcarbamate in higher plants and griseofulvin in the fungus Aspergillus nidulans. Inhibition of DNA synthesis in U. maydis and inhibition of cytokinesis in both organisms studied are secondary effects attributable to mitotic failure. A volatile compound toxic to U. maydis and S. cerevesiae was demonstrated in air over moistened benomyl. Hexane, through which such air samples were passed, or hexane extracts of aqueous suspensions of benomyl contained a substance having the same ir spectrum and retention time in two gas chromatographic systems as butyl isocyanate (BIC). BIC inhibited respiration of U. maydis and S. cerevesiae in a manner similar to benomyl. Benomyl partially inhibited respiration of subcellular particles from both organisms. MBC had essentially no effect on whole cell or subcellular particle respiration of either organism. Toxicity of benomyl preparations is not attributed to “benomyl” as such, but to two breakdown products, MBC and BIC. Therefore, differential effects of benomyl preparations and of MBC on growth and metabolism in fungi should be ascribed to BIC.