Studies were carried out to determine the effect of vitamins, temperature and pH on the biomass production by ectomycorrhizal fungi: <i>Laccuria bicolor, Hebeloma crustuliniforme, Suillus bovinus</i> and <i>Pisolithus arhizus</i>. Responses of particular organisms were different Three-factor ANOVA which enables the comparison of the effect of experimental factors on biomass yield has shown that the pH of the medium had a stronger effect on <i>L. bicolor, H. crustuliniforme</i> and <i>S. bovinus</i> than vitamins. Temperature was the factor exerting the strongest effect on the growth of <i>P. tinctorius</i>.