The invention discloses a cross-layer cooperative transmission protocol in a multi-hop wireless Ad Hoc network and an application thereof. In the protocol, an information receiver and a cooperative relay in one hop are determined at the same time, the receiver is closer to a target node, and the cooperative relay has a better channel condition. Furthermore, as an inter-flow signal reference problem caused by transmission of multiple information flows existing in the network is considered, an interference factor is introduced while the cooperative relay is selected in the protocol. As a result, the selected cooperative relay while transmitting information has relatively low interference to other information flows. The basic principle and method of the invention can be applied to a multi-hop multi-flow wireless Ad Hoc network in a Rayleigh fading environment.