The black tiger prawn, Penaeus monodon, is considered one of the world’s most commercially important decapod crustaceans. Despite the extensive progress in expanding P. monodon aquaculture, the species’ reproductive biology is far from understood; control of ovarian development and spawning is essential for a sustainable domestication program. One key challenge is the failure of ovarian maturation and spawning in most captive-bred female P. monodon, which requires compromising the eyestalk to secure seed supply. Since eyestalk ablation is deemed unfavorable due to production and animal welfare aspects, exploring cost-effective and efficient alternative(s) to replace eyestalk ablation requires a better understanding of female penaeid reproduction. An in-depth understanding of the intrinsic factors regulating ovarian development in P. monodon is a pre-requisite towards the development of a farm-applicable method to overcome ovarian developmental arrest in domesticated broodstock, a long-term goal for sustainable P. monodon aquaculture. To address the above challenge, we have set to characterize key molecular factors governing ovarian maturation in P. monodon using a multi-omics study, to develop and validate non-invasive tools for assessing ovarian stages in captive-bred P. monodon broodstock, and to develop in vitro assays for assessing factors impacting reproductive development in P. monodon broodstock without sacrificing them.