The aim of this paper is to analyze the reception of Spanish Renaissance sources containing knowledge about nature in the New World in the Czech Lands in the 16th and the beginning of the 17 century. Attention will be paid to Czech herbariums, cosmographical works and travel reports (e.g. Tadeáš Hájek, Sigismund of Púchov, Daniel Adam of Veleslavín or Adam Zalužanský of Zalužany), which provide information about products of nature of the New World, for example, based on the knowledge of works of C. Columbus, Mártir de Anghiera, N. Monardes or José de Acosta. The space of Czech castle libraries and cabinets of curiosities (especially the important Czech Renaissance dynasty of the Rosenbergs), which give evidence of the interest in Spanish Renaissance science and knowledge about the New World, will also be mapped. Based on the analysis of selected representative texts, illustrations and material objects (scientific or artistic), this paper will show the culturally contextual dependence and specificity of interpretation of Iberian natural-science knowledge about the New World in the Central European area.