Mitogen-activated protein kinase kinase kinase (MAPKKK, MAP3K) is located upstream of the mitogen-activated protein kinase (MAPK) cascade pathway and is responsible for receiving and transmitting external signals to the downstream MAPKKs. Although a large number of MAP3K genes play important roles in plant growth and development, and response to abiotic and biotic stresses, only a few members’ functions and cascade signaling pathways have been clarified, and the downstream MAPKKs and MAPKs of most MAP3Ks are still unknown. As more and more signaling pathways are discovered, the function and regulatory mechanism of MAP3K genes will become clearer. In this paper, the MAP3K genes in plants were classified and the members and basic characteristics of each subfamily of MAP3K were briefly described. Moreover, the roles of plant MAP3Ks in regulating plant growth and development and stress (abiotic and biotic) responses are described in detail. In addition, the roles of MAP3Ks involved in plant hormones signal transduction pathway were briefly introduced, and the future research focus was prospected.