Pietro Andrea Mattioli (1501–1577), the renaissance Italian physician, became famous primarily due to publishing his herbal. The Matiolli´s herbal is a typical example of the 16th century treatise on plants, which describes them especially for their medicinal purposes. It is primarily based on the ancient work of Dioscorides De Materia Madica but it also comprises the novel contributions from Mattioli and his contemporaries. The first edition appeared in 1544 in Mattioli´s mother tongue – Italian. The Latin version followed in 1554. It became very popular during the time of renaissance and it was translated into several languages. Mattioli also spent about ten years in Bohemia as a court physician of Archduke Ferdinand Tyrolský (1529–1596). Tree editions of the herbal were prepared during Mattioli´s stay in Prague: the Czech translation by Tadeáš Hájek z Hájku (1562, Prague), the German version by Georg Handsch (1563, Prague) and also the new Latin edition by Mattioli himself (1565, Venice). Later, in 1569, a new Czech edition was also printed, using some other herbals as sources too. The Mattioli´s herbal was very popular among Czech people. It remained a source of herbal medicine for a very long time as can be seen from the number of modern editions. This treatise will deal especially with the two Czech editions of Mattioli´s herbal and it will put them into the context of Czech botany. Other Mattioli´s publications, which originated during his stay in Prague, will be discussed as well. It will also focus on the translator of the first Czech edition: Tadeáš Hájek z Hájku (1526–1600), person, which is of a great importance for development of Czech science.