The application of Trichoderma spp. for the suppression of plant-parasitic nematode populations is a promising tool in biological control. Sixteen strains of six Trichoderma species (T. atroviride, T. harzianum, T. rossicum, T. tomentosum, T. virens and T. asperellum) were tested in vitro in order to identify the most appropriate strains to control the dagger nematode Xiphinema index. Mortality assays revealed that the strains of the widely investigated T. harzianum species have caused significant reduction of X. index populations, although T. harzianum strains were not the most efficient among all the tested fungi. Certain T. virens and T. atroviride strains and T. rossicum have triggered faster and higher mortality. Generally, our data indicate that Trichoderma species have innate ability to decrease X. index population. Furthermore, as we had difficulties with maintaining X. index in vitro, we successfully used a newly developed method to keep X. index specimens viable during the experiments.