Psilorhynchus homaloptera mainly inhabits turbulent waters in the lower reach of the Yarlung Zangbo River, Tibet, China, and it is the only species of Psilorhynchus in China. However, many important aspects of its biology remain poorly documented, which hinders the conservation efforts. Therefore, we studied its reproduction using 801 specimens from the Motuo sector of the lower Yarlung Zangbo River from December 2015 to November 2016. The sex ratio was 0.68?1 (male?female) for the overall population. Females reached a larger total length and weight (134 mm and 18.6 g) than males did (113 mm and 12.2 g). Lengths at maturity were estimated 80.0 mm for males and 93.1 mm for females. Our analyses of gonad development and the size distribution of oocytes suggest that P . homaloptera is a single spawner with the peak spawning period between September and October. Furthermore, the estimated mean absolute fecundity was 557±204 eggs per fish, and mean relative fecundity was 50.9±10.9 eggs per g of female weight. The fecundity of P . homaloptera increased linearly with increasing total length, total weight, ovary weight, and age. This study on the reproductive biology of P . homaloptera demonstrates that the fish was in a vulnerable state due to the increasing anthropogenic activities in the lower Yarlung Zangbo River.