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The systematics and distribution of South American sigmodontine rodents a matter of continuous revision and debate. The silky mice, genus Eligmodontia Cuvier, 1837, are among the most specialized murid rodents endemic to South America and its diversification for desert existence is associated with the uplift of the Andes and the early development of and Landscapes. Aiming to clarify the systematics of the species of silky mice occurring in the driest portion of the temperate Monte Desert in Argentina, qualitative and quantitative external and cranial characters, cytogenetics and molecular relationships, were studied. We characterized three karyotypes of Eligmodontia; two of which are described for the first time, and allocated them to previously named species. E. moreni Thomas, 1896 (2n=52 and FN=50), E. typus Cuvier, 1837 (2n=44 and FN=44) and E. marica Thomas, 1918. The later shows the same diploid number of E. typus, but its X-chromosome is not METACENTRIC but ACROCENTRIC. A discriminant analysis of external and cranial data separates E. moreni from E. typus and E. marica. Whereas these last ones show some degree of overlap. The morphological and chromosomal differentiation of Eligmodontia is sustained by DNA distances. Phylogenetic analyses show two major clades. One formed by E. moreni, E. puerulus and E. hirtipes, sharing a high FN and a northern distribution, and THE other formed by E. typus, E. marica and E. morgoni with Low FN and a central-southern distribution. Two centers of diversification are proposed to explain the evolution of Eligmodontia. (c) 2006 Deutsche Geseltschaft fur Saugetierkunde. Published by Elsevier GmbH. All rights reserved.