Volicitin (17-hydroxylinolenoyl-L-Gln) present in the regurgitant of Spodoptera exigua (beet armyworm caterpillars) activates the emission of volatile organic compounds (VOCs) when in contact with damaged Zea mays cv Delprim (maize) leaves. VOC emissions in turn serve as a signaling defense for the plant by attracting female parasitic wasps that prey on herbivore larvae. A tritiated form of volicitin was synthesized and shown to induce volatiles in the same fashion as the biological form. [3H]-L-Volicitin rapidly, reversibly, and saturably bound to enriched plasma membrane fractions isolated from Z. mays leaves with an apparent Kd of 1.3 nM and a Hill coefficient of 1.07. Analog studies showed that the L-Gln and hydroxy moieties of volicitin play an important role in binding. Treatment of plants with methyl jasmonate (MeJA) increased the total binding of [3H]-L-volicitin to the enriched plasma membrane more than threefold, suggesting that MeJA activates transcription of the gene encoding the binding protein. S. exigua feeding also increased total binding fourfold. Cycloheximide pretreatment of plants significantly decreased binding of radiolabeled volicitin to the enriched plasma membrane. These data provide the first experimental evidence that initiation of plant defenses in response to herbivore damage can be mediated by a binding protein-ligand interaction.