| Summary: | Artichoke (<i>Cynara cardunculus</i> L. var. <i>scolymus</i> (L.) Fiori) is a perennial plant of the Asteraceae’s family native to the Mediterranean basin. Italy has rich artichoke biodiversity, but many landraces are subjected to genetic erosion caused by increasing use of commercial varieties that are more homogenous in production. An Apulian landrace ‘Troianella’ was established in vitro to valorize and provide high-quality material for propagation in nurseries and, subsequently, for cultivation in production fields. The shoot proliferation was tested on four different growth media, adding cytokinin (-6-benzylamminopurine (BAP-0.05 mg L<sup>−1</sup>). Among these, the best results were achieved on MS519-A and BM media in which MS macronutrients were supplemented with additional doses of CaCl<sub>2</sub> and MgSO<sub>4</sub> (plus 120 mg L<sup>−1</sup> and 190 mg L<sup>−1</sup>, respectively). In vitro root induction was obtained with 10 mg L<sup>−1</sup> of Indole-3-acetic acid (IAA) and 30 g L<sup>−1</sup> of sucrose. Plants derived from tissue culture were acclimatized in greenhouse using mycorrhizal symbiosis to increase survival during the acclimatization phase and to improve their performance after transplanting in field. Three arbuscular mycorrhizal (AM) fungi (<i>Septoglomus viscosum</i>, <i>Funelliformis mosseae</i>, and Symbivit, a commercial mix) were added to a sterile substrate and compared to a sterile control without any AM fungal inocula. After 3 months, the best growth and plant appearance were on substrates with <i>S. viscosum</i> fungus or the commercial mycorrhizal fungi mix. The results supported a development of an efficient micropropagation protocol and a production of high quality plant material for sustainable farming of the endangered ’Troianella’ landrace.
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