The kiss1/gpr54 signaling system is considered to be a critical regulator of reproduction in most vertebrates. However this presumption has not been tested vigorously in non-mammalian vertebrates. Distinct from mammals, multiple kiss1/gpr54 paralogous genes (kiss/kissr) have been identified in non-mammalian vertebrates, raising the possibility of functional redundancy among these genes. In this study, we have systematically generated the zebrafish kiss1(-/-), kiss2(-/-) and kiss1(-/-);kiss2(-/-) mutant lines as well as the kissr1(-/-), kissr2(-/-) and kissr1(-/-);kissr2(-/-) mutant lines using transcription activator like effector nucleases (TALENs). We have demonstrated that spermatogenesis and folliculogenesis as well as reproductive capability are not impaired in all of these six mutant lines. Collectively, our results indicate that kiss/kissr signaling is not absolutely required for zebrafish reproduction, suggesting that the kiss/kissr systems play non-essential roles for reproduction in certain non-mammalian vertebrates. These findings also demonstrated that fish and mammals have evolved different strategies for neuroendocrine control of reproduction.