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The road network in Cyprus has seen an 88% increase in the last 20 years. This expansion has not been followed by any kind of assessment on the effects of the network on nature conservation. This is the first island-wide quantitative assessment of the size, character (surface types), pervasiveness and distribution of the road system with particular reference to Natura 2000 network on the island. We mapped roadless areas (i.e. areas at least one km away from nearest road) for the whole island and examined the spatial distribution with respect to Natura 2000. We tested the relationship between overall road density and road density of different road categories within terrestrial Natura 2000 sites to four zones which were defined on the basis of landform, principal land use and ownership. We employed three indices i.e. effective mesh size, splitting and division to measure fragmentation caused by the road network within Natura 2000 and investigated the relationship between road density and the above fragmentation metrics. Mean road density in Cyprus is 2.3 km/km2 which is comparable to road density values recorded in other Mediterranean countries such as France, Spain and Italy, which have much larger area and population. Roadless areas cover 4.5% of the island, and despite being scattered 80% is found within Natura 2000, which demonstrates the added value of the network for nature conservation. Road expansion has taken place throughout the island with the same intensity irrespectively of the zones examined. Fragmentation has been lower in sites on mountainous areas where sites are larger and under state ownership. Road density is negatively correlated (r = - 0.383, p = 0.05) with effective mesh size and positively correlated with both landscape division (r = 0.376, p = 0.05) and splitting index (r = 0.376, p = 0.05). Results corroborate that spatial configuration is an important property of the road network in addition to traffic load, length and density.With the shift from site based conservation to landscape level there is a challenge for integrating technical, human and ecological requirements into infrastructure planning.