| Summary: | Native tomato landrace varieties (<i>Solanum lycopersicum</i> L.) are sources of high genetic diversity and are adaptable to local environmental conditions. The in vitro propagation system can be used as a tool to produce disease-free, high-quality propagation material and preserve the unique characteristics of commercial Greek tomato varieties, such as the variety Areti and the traditional landrace variety Makedonia, both of exceptional quality and economic importance. Twenty- and 27-day-old cotyledon and leaf explants were cultivated in 4 regeneration media supplemented with 0 or 0.1 mg∙L<sup>−1</sup> indole-3-acetic acid and 0.5 or 1 mg∙L<sup>−1</sup> zeatin. Regeneration efficiency was dependent on variety, explant type and age, and regeneration media. Areti was a more appropriate target genetic material, exhibiting a six-fold greater response to regeneration compared to Makedonia, regardless of media and explant age. The regenerated shoots of both varieties were successfully rooted (60%) and acclimatized (98%). This regeneration protocol would be valuable in the production of propagation material for commercial and conservation practices and in breeding programs for genetic improvement.
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