The purpose of this study was to analyze the role of the incubation temperature of papuyu fish eggs (Anabas testudinius Bloch 1792) to obtain the ratio of female fish seeds. This research was conducted for 3 months starting from March 2019 until May 2019, at the Wet Laboratory of Lambung Mangkurat University, South Kalimantan Province. Fertilized Climbing Perch eggs are incubated at 20oC, 22oC, 24oC, and 26oC at 12 incubators (small box) with a stocking density of 25 eggs during incubation are observed and calculated the hatchability of eggs after the larvae hatch then cooling down the incubator is stopped. This study uses a completely randomized design (CRD) with 4 treatments and 3 replications, so that it will produce 12 experimental units. The results of analysis of variance (ANOVA) hatchability ranging from 90.67 to 100% had no significant effect. This study shows that the incubation temperature of papuyu fish eggs gives an effect on the female genital ratio of papuyu with the highest sex ratio in treatment C (24oC) of 100% and the lowest in treatment D (26oC) of 65.61% .The DMRT test results show that the sex ratio treatment C with D was significantly different also treatment A with D was different while other treatments were not significantly different.