The genus Narcissus has several endemic, rare and/or threatened species in the Iberian Peninsula and North Africa. In vitro propagation is a useful tool for threatened plants conservation used in ex situ strategies. Thus, the aim of this work was to study the propagation in vitro of bulb scale explants of five endemic, rare and/or endangered Narcissus species from the Iberian Peninsula, treated with different PGR combinations. Initiation was achieved in half-strength Murashige and Skoog (MS) basal salts and vitamins, 10 g/L sucrose, 500 mg/L casein hydrolysate, 2 mg/L adenine, 10 mg/L glutathione and 5.5 g/L plant agar. In the multiplication phase, the highest bulblet proliferation was obtained in MS medium supplemented with 30 g/L sucrose and the combination of 10 μM 6-Benzylaminopurine (BAP) + 5 μM α-Naphthaleneacetic acid (NAA) in N. alcaracensis, N. eugeniae and N. hedraeanthus; 20 μM BAP + 5 μM NAA in N. jonquilla and N. yepesii. The highest rooting was obtained with 5 μM NAA + 1 μM Indole-3-butyric acid (IBA) for all species (>75%) and more than 80% of the produced bulblets were successfully acclimatized.