This study was designed to evaluate different substrates and temperatures on the germination of Cnidosculus phyllacanthus seeds in the laboratory, to define a protocol for the species to be used in the rules for testing seeds. The constant temperatures of 25ºC and 30ºC and alternated temperatures 30-35º, 20-30º and 25-35ºC combined with substrates of filter paper, sand, vermiculite and towel paper, moistened with nistatina solution 0,2% were evaluated.