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Segmentation of landholders has rarely been undertaken in the natural resource management literature, yet it provides a rich set of information for planners to improve targeting of public investment and resources. Segmentation allows the identification of segments of landholders that are ‘investment ready’, their location and characteristics, and the land conservation incentive programmes that they would be most willing to participate in. Segments that are not investment ready are also identified, and the instances where they reside in critical ecological habitats. Further, segmentation analysis provides insight into constraints to participation among these segments. We report the results of a large-scale segmentation exercise involving qualitative research and a quantitative survey of 5904 landholders in Australia. Using several new constructs related to human capital, trust and social connectedness not previously used for segmentation, we find three mainstream and two lifestyle segments which differ substantially in their socio-demographics and their propensity to be involved in incentive programmes.