Until today, two important questions about the population genetic structure and parental assignment remained unclear: (1) What is the degree of genetic differences among the Hilsa shad populations available in different migratory routes of the species? (2) Are the juveniles of Hilsa shad (locally known as Jatka) of a particular habitat return to their respective natal rivers (rivers where they were born/nursed) for spawning as adults? To unravel the answers of the above two questions, the present study was conducted by identifying a set of neutral and adaptive genetic markers. The NextRAD sequencing technique was employed to genotype 300 individuals of the species collected from nine strategic aquatic habitats including sea, estuary and different rivers for revealing parental assignment and stock structure of the Hilsa shad population in Bangladesh waters. We applied the FST OutFLANK and pcadapt approach to determine adaptive vs. neutral SNP loci, and additional analysis were employed to verify the significance of these putatively adaptive loci under selection in the context with different migratory routes and habitats.