Development of plant in vitro regeneration protocols has significantly contributed to research investigations and rapid propagation in commercial scale. However, due to considerable variations among plants, the technique encounters various hurdles. Major challenges are encountered during the micropropagation of woody plants, due to their range of secondary metabolites. Tissue and media browning, systemic contamination, vitrification, shoot tip necrosis (STN), tissue fasciations, somaclonal variations are some of the commonly encountered problems. Researchers in the recent years introduced various novel and unique protocols to eradicate these challenges, like nanosilver mediated sterilization, various new culture media for woody plant propagation, discovery of many new natural PGR having better properties like tapolins, diethyl aminoethyl hexanoate (DA-6), thidiazuron (TDZ) etc. have contributed immensely to the development of the subject. Although the basic concept of plant tissue culture remains same, the interpretation is subjected to development from time to time.