The survey of occurrence of six chosen virus pathogens and phytoplasmas complex was done on plants of grapevine (Vitis vinifera L.) which showed symptoms of short-shoot syndrom. The results of serological and molecular tests did not confirm either virus or phytoplasma infection as the main source of short-shoot syndrom. The presence of 6 viruses in samples taken from 45 affected plants of grapevine on 4 habitats. The highest occurrence of viruses was found out on habitat Moravská Nová Ves, where all taken samples were infected by Grapevine leafroll-associated virus GLRaV-1 (100 %). In 66 % of the samples taken from that habitat were detected mixed infection of Grapevine virus A and GLRaV-1. These 2 pathogens but were not detected in samples from affected plants from the other habitats or very sporadically – just in 2 plants. Another 4 virus pathogens were detected either sporadically (Grapevine fleck virus, Grapevine leafroll-assoc. virus GLRaV-3, Arabis mosaic virus) or not at all (Grapevine fanleaf virus). From 270 tests made to 6 viruses were only 20 positive, e. g. 7.4 %. It means that from 45 plants were 15 infected at least by 1 virus (33 %). The phytoplasmas complex was tested in 28 plants. The result was positive only in 1 plant, by another test the Potato stolbur phytoplasma was confirmed.