This paper introduces ecosystem services (ES) assessment methods in the 7th Framework Programme of EU OpenNESS project in our Trnava city case study. The main objective of the project was operationalisation of the Natural Capital and Ecosystem Services concepts by developing conceptual and regulatory frameworks and evaluation method and testing these in 27 case studies of 17 countries. The main part of this paper is devoted to introducing the ES assessment applied in the Trnava case study, especially linked to landscape and spatial planning. Ten relevant ES were selected by experts and stakeholders and these were evaluated by four methods based on GIS procedures and stakeholder participation. The basic Spreadsheet method focuses on reclassification of current land cover applied in comparing municipal and regional area capacity for ES provision. We then employed the GreenFrame method with analytic maps providing detailed spatial data sets on landscape quality. These are expressed as spatial themes similar to those used in expressing landscape capacity for ES provision. Although this method is more demanding for input data, it is more relevant for landscape planning processes. The QuickScan method used is designed for participative decision-making, involving mainly planners, decision-makers and other respective stakeholders. While this method requires the most detailed ecosystem spatial information, it contributes significantly to urban and spatial planning processes. The last utilised method, ESTIMAP, provides a set of specific models for individual ES evaluation. Our case study evaluated recreational ES using combined selected spatial data and a questionnaire survey. The results of our research confirm our conclusion that suitability of using particular and combined ES assessment methods depends strongly on the availability of existing inputs, the purpose of the evaluation process and expected purpose of use of results.