Initially mycoheterotrophic plants have recently been declining in the wild, even without apparent causes. They are affected by a number of biotic and abiotic factors. The aim of this work is to investigate how selected factors, such as nitrate or symbiotic fungi, may influence their distribution in nature. In particular, the work applies in vitro experiments, molecular determination of fungal symbionts and stable isotope analyses. It describes both the effects of abiotic factor, specifically nitrate, and biotic interactions of initially mycoheterotrophic plants with fungal symbionts. The inhibition of germination by extremely low concentrations of nitrate in asymbiotic in vitro cultures was observed in several orchid species. The degree of sensitivity of each species to nitrate corresponds with the nitrate content of the soil and the nutrient availability requirements of the species according to Ellenberg indicator values. The inhibitory effect of nitrate on orchid germination was also observed in symbiotic in vitro cultures. Out of five tested fungal strains, only one Ceratobasidium was capable of eliminating the inhibitory effect of nitrate. Furthermore, the work reveals that green mixotrophic orchids use photosynthates to nourish the aboveground parts, whereas the belowground parts are...