The invention relates to a rhinogobio ventralis manual propagation technology which specifically includes four steps of domesticating, strengthening propagation, parent fish spawning induction, fertilization incubation and the like. The method breaks through the rhinogobio ventralis manual propagation technology for the first time at home and abroad, successfully solves the technical difficulties of high stress, difficult domesticating and streak gonads in the rhinogobio ventralis domesticating propagation process, and successfully achieves rhinogobio ventralis manual propagation for the first time. In the period from June 27th to May 9th in 2014, spawning induction of rhinogobio ventralis is conducted successfully for five times, about 97 thousand fertilized eggs are obtained, the fertility rate reaches 98.9%, about 30.5 thousand new-born fish fries are obtained, and stock enhancement of the rhinogobio ventralis becomes possible. The method achieves the technical reference and demonstration effects on protection of valuable rare fishes in the upstream of the Yangtze River and has great economical benefit, social benefit and ecological benefit.