The study tested the influence of the low pH values of the nutrient medium for rooting on the growth and development of the shoots of five white poplar (Populus alba L.) genotypes. After 35 days of in vitro cultivation following characters were measured: height of shoot, the number of roots, length of the longest root, the percentage of survival and rooting. Three different pH values of the medium: 3.0, 4.0 and 5.5, buffered by sodium citrate buffer, and standard medium for rooting with pH 5.5 without the addition of citric acid (control), sterilized in microwave oven, were used. Generally, a statistically significant lower values in all studied morphological parameters have been recorded on the media with pH 3.0, compared to all the other tested media. The highest values of all investigated parameters were observed in clone L-80. The results indicate that the in vitro test culture might help to narrow the selection of white poplar genotypes that could be used on acid soils.