The genetic variability found in tree species presents an opportunity for enhancing genetic traits and utilizing superior genotypes through clonal forestry. While traditional macropropagation methods have limitations in mass-producing clonal plants for challenging-to-root tree species, plant tissue culture-based biotechnology emerges as a solution to overcome these challenges and complements tree improvement programs. There are three main methods of micropropagation for tree species: axillary shoot proliferation, adventitious shoot regeneration (both direct and through the callus phase), and somatic embryogenesis (both direct and through the callus phase). Among these, axillary shoot induction is the preferred and safest method for producing genetically true-to-type planting material. Direct adventitious regeneration and somatic embryogenesis follow in preference. However, the presence of prolonged cultures (over 1-year old) may pose challenges of genetic variability. Therefore, before large-scale micropropagation for operational planting, it is crucial to conduct genetic fidelity studies using DNA markers to ensure a secure mode of regeneration and the maintenance of genetic uniformity in planting materials. Several factors significantly influence micropropagation, including (1) the type and source of explant (seedling/mature tree), (2) the management of trees serving as the explant source, (3) the timing of explant collection, (4) surface sterilants, their concentrations, and treatment duration, (5) nutrient media, (6) growth hormones and their concentrations, (7) genotypes/clones, (8) growth adjuvants, (9) gelling agents and media pH, (10) carbohydrate source and its concentrations, (11) incubation conditions of cultures, and (12) hardening conditions and duration. Micropropagation, requiring a small quantity of plant material (explant), offers high production potential for mass multiplication of selected genotypes/clones and plant rejuvenation. It can be carried out in small spaces, allowing year-round production unaffected by external environmental factors. The benefits include uniformity of planting material, high production rates, and germplasm preservation. This chapter provides detailed insights into the micropropagation of tree species through various modes of plant regeneration.