Abstract A total of 320 accessions of subterranean clover (Trifolium subterranenum L.) were screened for adaptation to the environmental conditions in southern Germany, as well as their suitability as cover crops or living mulches. The accessions, derived from collections of the Department of Agriculture and Food Western Australia and the International Research Center for Agriculture in Dry Areas, were propagated in the greenhouse and grown in rows. Selected accessions were then tested in plots and as an intercrop in wheat, in order to test their suitability as living mulch. Moreover, a frost resistance screening was carried out in a climate chamber. Adaptation to southern German conditions and frost tolerance was generally better than expected, as most accessions survived winters with snow cover and transitory temperatures as low as -10∘C. Accessions with particularly high frost tolerance were identified and several accessions persisted over four generations of self-reseeding. Although there was a large range of morphological characteristics, no differences concerning their suitability as living mulch could be observed. The results suggest that subterranean clover is sufficiently winter hardy to be grown as an overwintering cover crop or living mulch in southern Germany. Commercially available cultivars appear suitable in the first instance, even though these have not been selected for cold winters. However, adaptation might be further improved by targeted selection among accessions.