Tigers Panthera tigris face a wide and complex array of threats. Given limited time and resources it is essential to direct conservation actions based on the relative importance of each threat. The Sundarbans Reserve Forest is the last stronghold of tigers in Bangladesh and supports one of the largest populations of tigers in the world. As in other tiger landscapes, the threats faced by the tigers have yet to be assessed. This study follows an approach developed by The Nature Conservancy to identify and prioritize threats and set a time-frame for their reduction. We identified a total of 23 threats; four were linked to tigers, two to prey and 17 to habitat. Of the identified threats, the highest ranked included poaching of tigers, poaching of prey, sea-level rise, upstream water extraction/divergence, wood collection, fishing, and harvesting of other aquatic resources. All threats were then scheduled for reduction, based on the rank and current information base for each threat and the likely time-frame for implementing potential solutions. This study demonstrates how the application of a prioritization framework can greatly improve the focus and likelihood of success of any species- or ecosystem-based conservation programme.