Involvement of pH and IBA on adventitious root initiation was investigated with Malus domestica Borkh. microcuttings. The pH of unbuffered root initiation medium (RIM) increased from 5.6 to 7 within 2 days. Buffering with 2[N-morpholino] ethanesulfonic acid (MES) adjusted to specific pHs with potassium hydroxide prevented pH changes and resulted in a 2-fold higher root count at pH 5.5 compared to pH 7 or unbuffered medium. As pH decreased, lower concentrations of IBA were required to increase root counts. Colorimetric measurement of IBA in buffered RIM showed greater IBA loss and higher root count were associated with lower pH levels in all cultivars. This suggests that IBA loss from RIM depends on medium pH, which affects root count. Root count differences between easy-to-root through difficult-to-root cultivars were not consistent with amount of IBA loss from RIM. Cultivar differences in root count could not be explained solely by IBA loss from RIM.