Regular mowing of grassland is often necessary for plant conservation, but uncut vegetation is needed by many arthropods for overwintering. This may lead to conflicting management strategies for plant and arthropod conservation. Rotational fallows are a possible solution. They provide a spatio-temporal mosaic of mown and unmown areas that may combine benefits to both plants and arthropods. We tested if rotational fallows enhance spider overwintering in fen meadows. Rotational fallows consisted of three adjoining strips 10 m wide and 35–50 m long. Each year, one of these strips was left unmown (fallow) in an alternating manner so that each strip was mown two out of three years. Spiders were sampled during spring with emergence traps in nine pairs of currently unmown fallow strips and completely mown reference plots. Fallows significantly enhanced orb-weavers (Araneidae), sac spiders (Clubionidae) and ground spiders (Gnaphosidae). However, only 4.7% of the total variation in community composition was attributable to fallows. Community variation was larger between landscapes (34.5%) and sites (38.2%). Also β diversity was much higher between landscapes (45 species) and sites (22 species) than between fallows and mown reference plots (10 species). We conclude that the first priority for spider conservation is to preserve as many fen meadows in different landscapes as possible. Locally, rotational fallows enhance overwintering of the above-mentioned spider families, which are sensitive to mowing in other grassland types as well. Thus, rotational fallows would probably foster spider conservation in a wide range of situations. However, stronger effects can be expected from larger and/or older fallow areas.