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Biodiversity surveys for myxomycetes (Amoebozoa) were carried out in three consecutive years (2009 to 2011) in the cold arid Patagonian Steppe, Argentina. The surveys, the first to cover such an extensive area in South America, form part of the Myxotropic project funded by the Spanish Government. Specimens were collected in 174 localities in four different provinces (Neuquen, Rio Negro, Chubut and Santa Cruz), between 36 degrees and 52 degrees S latitudes. The most common types of substrate investigated were the dominant shrubs and grasses of the Patagonian steppe, and the Nothofagus forests, characteristic of the transition areas, but other plants such as small cacti and cushion plants were also included in the survey. A total of 133 different species and 5 varieties of myxomycetes representing 31 genera were identified in the 1134 specimens collected either in the field, or from moist chamber cultures prepared with samples of plant material obtained from the same collecting sites. The results include one species new to science, Perichaena nigra, and 17 species and two varieties that were previously unknown for either the Neotropics or South America, Badhamia armillata, Dianema mongolicum, Didymium annulisporum, D. leptotrychum, D. orthonemata, D. sturgisii, Echinostelium coelocephalum, Licea deplanata, L. nannengae, Macbrideola argentea, M. oblonga, Oligonema aurantium, Perichaena luteola, P. madagascariensis, Physarum luteolum, Protophysarum phloiogenum, Trichia contorta var. attenuata, T. contorta var. iowensis, T. erecta. An additional 19 species are new records for Argentina. These additions make Argentina the country in South America, at present, with the greatest number of myxomycetes catalogued having more than 50% of the species cited from the whole Neotropics. Diversity and biogeographic distribution of these organisms are discussed, and taxonomic comments on rare or unusual species are included and illustrated with photographs by LM and SEM. The results indicate that the myxomycetes, are widely distributed and are a normal component of Patagonian biota. Many of the substrates investigated were endemic plants from the region and are new substrates for a number of species of myxomycete. Differences between the variety of species in this area and others in Argentina and Chile, suggest a certain regional specialization of these organisms, the assemblage of which appear to depend on plant substrate species.