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Propagation of horticultural plants through in vitro culture has become a viable alternative to conventional propagation methods worldwide. Rapid production of true-to-type and disease-free plants in controlled laboratory conditions, in a small space, in a short time, and regardless of seasonality are especially desired for producing cuttings for commercial purposes in horticulture. On the other hand, in vitro culture is an important tool in plant breeding, as well as in plant conservation and long-term storage. This reprint contains an anthology of studies regarding the latest advancements in the field of in vitro culture of horticultural plants. Readers will gain a deeper understanding of how this technology is shaping the future of crop improvement and horticulture. The reprint not only highlights the remarkable progress made in in vitro technology, but also emphasizes ongoing efforts to overcome issues related to somaclonal variations, virus elimination, and acclimatization of micropropagated plants.