Micropropagation has become a reliable and routine approach for large-scale rapid plant multiplication, which is based on plant cell, tissue and organ culture on well defined tissue culture media under aseptic conditions. A lot of research efforts are being made to develop and refine micropropagation methods and culture media for large-scale plant multiplication of several number of plant species. However, many forest and fruit tree species still remain recalcitrant to in vitro culture and require highly specific culture conditions for plant growth and development. The recent challenges on plant cell cycle regulation and the presented potential molecular mechanisms of recalcitrance are providing excellent background for understanding on totipotency and what is more development of micropropagation protocols. For large-scale in vitro plant production the important attributes are the quality, cost effectiveness, maintenance of genetic fidelity, and long-term storage. The need for appropriate in vitro plant regeneration methods for woody plants, including both forest and fruit trees, is still overwhelming in order to overcome problems facing micropropagation such as somaclonal variation, recalcitrant rooting, hyperhydricity, polyphenols, loss of material during hardening and quality of plant material. Moreover, micropropagation may be utilized, in basic research, in production of virus-free planting material, cryopreservation of endangered and elite woody species, applications in tree breeding and reforestation.