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Synergies between biodiversity conservation objectives and ecosystem service management were investigated in the Succulent Karoo biome (83,000 km(2)) of South Africa, a recognised biodiversity hotspot. Our study complemented a previous biodiversity assessment with an ecosystem service assessment. Stakeholder engagement and expert consultation focussed our investigations on surface water, ground water, grazing and tourism as the key services in this region. The key ecosystem services and service hotspots were modelled and mapped. The congruence between these services, and between biodiversity priorities and ecosystem service priorities, were assessed and considered in relation to known threats. Generally low levels of overlap were found between these ecosystem services, with the exception of surface and ground water which had an 80% overlap. The overlap between ecosystem service hotspots and individual biodiversity priority areas was generally low. Four of the seven priority areas assessed have more than 20% of their areas classified as important for services. In specific cases, particular service levels could be used to justify the management of a specific biodiversity priority area for conservation. Adopting a biome scale hotspot approach to assessing service supply highlighted key management areas. However, it underplayed local level dependence on particular services, not effectively capturing the welfare implications associated with diminishing and limited service provision. We conclude that regional scale (biome level) approaches need to be combined with local level investigations (municipal level). Given the regional heterogeneity and varied nature of the impacts of drivers and threats, diverse approaches are required to steer land management towards sustainable multifunctional landscape strategies.