Culture Media for Woody Plant Tissue Culture: From Classical Formulations to Rational Optimization

Plant tissue culture is essential for clonal propagation, germplasm conservation, genetic transformation, and genome editing in woody plants, yet these species remain highly challenging in vitro due to developmental recalcitrance, phenolic oxidation, endogenous contamination, seasonal effects, hyperhydricity, rooting difficulty, and strong genotype dependence. Medium formulation is therefore often a decisive factor rather than a routine technical step. This review examines the development, composition, and application of tissue culture media for woody species, emphasizing how specialized formulations emerged to overcome limitations of general-purpose media such as Murashige and Skoog medium. Major woody-targeted media, including Woody Plant Medium, Quoirin and Lepoivre medium, Anderson’s medium, Driver and Kuniyuki Walnut medium, Rugini Olive Medium, Litvay medium, and DCR medium, are discussed in relation to the physiological needs of their target species. Key medium components are reviewed, including mineral nutrients, carbon sources, vitamins, plant growth regulators, organic additives, gelling agents, and pH, with attention to nitrogen form, ionic strength, calcium supply, iron availability, and pathway-dependent plant growth regulator selection. The review further highlights applications in conservation, micropropagation, transformation, and genome editing, and discusses emerging approaches such as statistical optimization, machine learning, nanoparticles, temporary immersion systems, biostimulants, and morphogenic regulators.