The success in all of plant tissue culture protocols is related to decontamination of explants. The purpose of the current study was to evaluate new compounds for the control of external and internal contamination and their impact on different grape cultivars in process of tissue culture. In this study leaf explants of grape Vitis vinifera L. (cv. "Rashe","Khoshnave" and "Farkhi") was used. Silver nanoparticles used for disinfection of explants surface in the process of tissue culture. In this study ethanol 95 % for 30 seconds + silver nanoparticles (100, 500 and 1000 ppm) used for surface disinfection during 10 and 20 minutes. The results showed low rates of burned explants in nanosilver treatments. Generally the explants tissues had high level of tolerance to silver nanoparticles. Silver nanoparticles treatments had moderate and high effect on bacterial contamination control, but had moderate effect for fungi contamination control. The best result was obtained with increasing concentrations of silver nanoparticles and time of disinfection.