Centaurea L. is a large genus which comprised of several species, many of which are used in folk medicine. Centaurea paxorum Phitos & Georgiadis (Asteraceae) is a range-restricted Greek endemic only to a single island (Paxoi-Antipaxoi) and is assessed as vulnerable according to the IUCN criteria. Tissue culture techniques have been established as a useful approach for ex situ conservation of rare, endemic or threatened plant species. This report describes the micropropagation of C. paxorum, an extremely rare-endangered indigenous plant species endemic to the Greek flora, as a conservation measure which does not cause damage to the wild plants used as explant source. There are no studies identified aiming to propagate this species in vitro. The effect of the cytokinin; BA applied at 0.1 mg/l in combination with 3 different auxins (IBA, NAA, IAA) each applied at 0.025 mg/l on multiple shoot induction potential was studied. Induction of adventitious shoot regeneration from shoot tips was achieved approximately 100% with the production of 4.83 shoots per explant of average mean length of 15.51 mm after 30 days of culture in MS medium supplemented with 0.1 mg/l BA+ 0.025 mg/l IBA. In the subsequent rooting stage, the individual effect of the 3 auxins (IBA, NAA, IAA) each applied at 0.1 mg/l was studied. The elongation of the initial shoot tips was best enhanced (41.67 mm) with the application of 0.1 mg/l NAA to the MS rooting medium, being increased approximately by 1.5 cm compared to the control explants (auxins-free). Regenerated adventitious shoots were best rooted on MS medium containing 0.1 mg/l IAA (30 days) exhibiting 100% rooting with the formation of 8 roots per rooted microplant 43.26 mm long. The present study shows the results on the regeneration capability of C. paxorum in tissue culture. This study is the first to record an efficient protocol to mi­cropropagate C. paxorum using shoot-tip explants.