The pathogenicity and growth of six cultures of the bark-beetle-associated blue-stain fungus Ceratocystis polonica were compared. Four were subcultures, established at different times, of the same isolate (80–53/7), and at least one of these was suspected to have lost its pathogenicity to Norway spruce (Picea abies) during subculturing. Two other cultures (93–208/115 and 94–169/13) were pathogenic. The pathogenicity of all cultures to Norway spruce was compared in three inoculation experiments, using (i) massive inoculation (≈144 inoculations per tree), (ii) low-density inoculation (14 inoculations per tree) of larger trees and (iii) single inoculation of 2-year-old seedlings. Fungal growth rates were tested in vitro on malt agar, and on malt agar under oxygen-deficient conditions. Isolates 93–208/115 and 94–169/13 and one of the subcultures of isolate 80–53/7 were pathogenic in all three inoculation experiments. Two other subcultures were nonpathogenic, while the last subculture was intermediate. The pathogenic cultures had higher growth rates on malt agar than the nonpathogenic and intermediate cultures, and they also grew better under oxygen-deficient conditions. These results demonstrate that blue-stain fungi may lose their pathogenicity after serial vegetative transfers on artificial substrates. The results are also consistent with the hypothesis that rapid growth and the ability to grow in oxygen-poor environments are important pathogenicity factors in blue-stain fungi.