Sensitivity to substrate-borne vibrations is widespread in animals and evolutionarily precedes hearing but, compared with other sensory modalities, we know little about vibrational communication, particularly in vertebrates . For plant-dwelling arthropods, vibrations are likely as important as sound [1-3]. Arboreal vertebrates excite plant vibrations with most movements , but the behavioral relevance of these vibrations has not been tested experimentally [5, 6]. In playback experiments using a robotic model frog and an electrodynamic shaker, we demonstrate that plant-borne vibrations generated by the shaking (tremulation) display of male red-eyed treefrogs (Agalychnis callidryas) are a vibrational signal, necessary and sufficient to elicit tremulations in response. A trend toward increased aggression during visual playbacks suggests that the visual component of tremulations may also convey information. In male-male contests, tremulations were the most frequent aggressive display, and their use and vibrational characteristics varied with male size and conflict context. Nearly all of A. callidryas' signaling behaviors, including tremulations and acoustic calls, excite strong, stereotyped vibrations that travel through plants and could be informative to receivers. Our results demonstrate that vibrational signals serve a key role in the biology of one well-known arboreal frog and suggest that consideration of the vibrational modality may significantly broaden our appreciation of the behavior and evolution of arboreal vertebrates.