The aim of this study was to characterize karyotypes of central European spiders of the genera Arctosa, Tricca, and Xerolycosa (Lycosidae) with respect to the diploid chromosome number, chromosome morphology, and sex chromosomes. Karyotype data are reported for eleven species, six of them for the first time. For selected species the pattern in the distributions of the constitutive heterochromatin and the nucleolar organizer regions (NORs) was determined. The silver staining technique for detecting NORs of lycosid spiders was standardized. The male karyotype consisted of 2n = 28 (Arctosa and Tricca) or 2n = 22 (Xerolycosa) acrocentric chromosomes. The sex chromosome system was X1X20 in all species. The sex chromosomes of T. lutetiana and X. nemoralis showed unusual behaviour during late diplotene, namely temporary extension due to decondensation. C-banding technique revealed a small amount of constitutive heterochromatin at the centromeric region of the chromosomes. Two pairs of autosomes bore terminal NORs. Differences in karyotypes among Arctosa species indicate that the evolution of the karyotype in this genus involved autosome translocations and size changes in the sex chromosomes. Based on published results and those recorded in this study it is suggested that the ancestral male karyotype of the superfamily Lycosoidea consisted of 28 acrocentric chromosomes. and Petr DOLEJŠ, Tereza KOŘÍNKOVÁ, Jana MUSILOVÁ, Věra OPATOVÁ, Lenka KUBCOVÁ, Jan BUCHAR, Jiří KRÁL.