The rural land use sector could potentially mitigate a large amount of GHG emissions. Implementation requires the engagement of farmers and other land managers. Understanding the barriers and enablers for the uptake of these practices is essential both to inform policy-makers and to achieve effective policy outreach. In Scotland, the rural land use sector is subject to a greenhouse gas (GHG) emission reduction target of 21% by 2020 relative to 1990 levels. This study contributes to the body of research on stakeholders’ perspectives about suitability of climate change mitigation practices at the regional level. Mixed-methods were used to collect the data, namely participatory workshops with scientists and relevant stakeholders, a farmer questionnaire, and focus groups with farmers. Findings show that farmers were mainly willing to expand the uptake of mitigation practices they were already implementing because they consider these are the most cost-effective. Barriers to the implementation of mitigation practices are mainly related to physical–environmental constraints, lack of information and education and personal interests and values. Similarly, enablers are also related to physical–environmental factors and personal interests and values. Economic incentives, voluntary approaches and provision of information have been identified by workshop participants as the most favourable approaches needed to promote the uptake of technically feasible mitigation practices. Farmers, however, consider that agriculture is a “special case” and should have not to comply with GHG emission reduction targets. Mitigation practices, should, therefore, be integrated with other initiatives.