The structure and quality of the adventitious root system formed on a microcutting stem is crucial to the successful acclimation, survival, and ultimate performance of micropropagated plants. Despite increasing evidence that the rooting method impacts on the character of the framework root system, very little research has elucidated the consequences of standard rooting methods on plant quality and performance. Root initiation on microcuttings is handled by a wide range of strategies in commercial practice. In comparison to in vitro roots, ex vitro roots have smaller root diameters, larger vascular diameters, greater length, more branch root development, and root hairs. One important microenvironmental parameter - light - was investigated to determine its contribution to root character. Typically, in vitro root systems are exposed to light throughout the root initiation period. Parallel treatments were established of in vitro light and dark rooting and ex vitro rooting of four woody species. Regardless of light exposure, the overall diameter of adventitious roots was larger for in vitro treatments than the ex vitro treatment. Vascular development was significantly more advanced ex vitro. These results suggest that light is not a major influence contributing to the differences between ex vitro and in vitro root character.