The present invention relates to a timeslot multiplexing wireless chain multi-hop cross-layer optimization method, which is applicable to wireless long-distance chain multi-hop communication. A chain multi-hop timeslot allocation algorithm is designed by combining a TDMA channel access mechanism with an AODV routing protocol, and a transmit-receive timeslot is allocated to each node, to ensure that a same timeslot is allocated to any two nodes outside a two-hop neighbor node range, and implement cross-layer optimization. Because neighbor nodes in a two-hop range easily conflict, three timeslots need to be allocated for data transmission. A transmit-receive timeslot of a node is allocated by using a routing control message of a network layer without adding overhead of another control message. According to the optimization method, a channel waste problem caused by conventional TDMA is resolved in a timeslot deployment manner by using a cross-layer design method and comprehensively considering a TDMA network feature.