This study provided new knowledge on reproductive traits of the unique mudskipper Periophthalmodon septemradiatus that is also a potential aquarium pet. Fish specimens were obtained by using fishing rods at the muddy flat in tributaries of the Hau River from August 2017 to July 2018. Data analysis of a collection of 1,504 individuals (930 males and 674 females) showed that the sex ratio of this species was significantly different from the ratio 1:1. As mature gonads were found monthly during the study period, it is suggested that P. septemradiatus is an iteroparous species releasing eggs for a year‐round cycle. The fish length at first mature (Lm) showed an increase from estuary to upstream areas. Male P. septemradiatus had higher Lm values than that of females. Likely, the batch fecundity increased from estuary region (5,916 ± 312 SEM eggs) towards upstream area (11,451 ± 953 SEM eggs). The fish total length and weight values had strong positive relationships with batch fecundity, as shown by high determination values (r2), suggesting that batch fecundity increased with fish size. This study provided more information on reproductive biology of P. septemradiatus, which can be utilized for fish population conservation and artificial reproductive study.