Autochthonous cultivars Plavac mali, Maraština, Pošip i Škrlet are of economical importance of Croatian viticulture but up to this time they have been neglected in viticulture production. In the last decade there has been an increased interest in preserving the autochthonous cultivars so the process of economical revitalization had been started. In cultivars Plavac mali, Maraština, Pošip i Škrlet is recorded high rate of virus infection, yet there is a low number of healthy vines – in some cultivares the healthy vines have not been found. Therefore the need for recovery from virus infections and production of healthy plant material have occurred. To stop the spread of virus infection the tissue culture in vitro is applied and it is considered most efficient technique of obtaining virus free plant material. In Croatian cultivars there is an infestation of GLRaV­1, 3(Grapevine leafroll associated virus) which cause leafroll and GFLV(Grapevine fanleaf virus) which causes fanleaf in grapevine. The research includes healthy genotypes of cultivares Plavac mali, Maraština, Pošip i Škrlet and genotypes infected with GLRaV1, 3 and GFLV. The technique of micropropagation of nodal segments is applied which means the nodal segments have been inoculated on culture media ½ MS . Two and four weeks after the inoculation the observation and measuring were performed. Two weeks after an inoculation the culture establishment was observed – the rate of inoculated nodal segments in relation to appearing of plantlets, and four weeks after the inoculation the measuring of was performed: the length of appearing plantlets and number of nodes. The culture media ½ MS showed good results for in vitro growth of cultivares Plavac mali, Maraština, Pošip i Škrlet. The best success in culture establishment reached the cultivares Plavac mali i Pošip and lower the cultivares Maraština i Škrlet. Plavac mali has achieved the highest length and number of nodes, the lowest length achieved Pošip, and Maraština and Škrlet have achieved the smallest number of nodes.