109 Revista del Jardín Botánico Nacional 30-31: 109-111, 2009-2010 In vitro conservation of Angelica pachycarpa, an Iberian endemic Apiaceae of the Portuguese Berlenga Islands Ana Cristina Pessoa Tavares*, Lígia Salgueiro** & Jorge Canhoto* *Jardim Botânico, Universidade de Coimbra. Portugal. **Laboratório de Farmacognosia/CEF, Universidade de Coimbra. Portugal. INTRODUCTION Continuing our research on the in vitro and ex situ conservation of the Iberian endemic Apiaceae in Portugal (Tavares & al. 2007), we present the first results on the micropropagation of Angelica pachycarpa Lange. This species requires particular conservation measures since its geographical distribution in Portugal is resctrited to very small areas of the west coast in the north of Lisbon and in the Berlenga Islands (Parker, 1981; Castroviejo & al., 2003; Gimenez & al. 2004). For a long time, plants from the Apiaceae family have been used as spices or drugs and herbal medicinal products from this botanic family are actually described in some Pharmacopoeias, as having antiseptic, expectorant, diuretic, carminative, vasodilator, or spasmolytic actions (Ekiert 2000; Nalawade & al. 2003). As other members of the Apiaceae family, Angelica L. is an essential oil producing plant and is largely cultivated in Europe due to its flavouring qualities for food and liquor production (Eeva & al. 2003) and also for the anti-oxidant properties of the essential oils (Wei & Shibamoto 2007). Most of the pharmaceutical industry is highly dependent on wild population for the supply of raw materials for extraction of medicinally important compounds. Due to a lack of proper cultivation practices, destruction of plant habitats, and illegal and indiscriminate collection of plants from these habitats, many medicinal plants are severely threatened. Advanced biotechnological methods of culturing plant, cells and tissues should provide new means of conservation and propagation of valuable, rare and endangered medicinal plants (Nalawade & al. 2003). Somatic embryogenesis and other techniques of micropropagation provide a means for large scale plant production and seems to be an ideal method to achieve the multiplication of species for purposes of plant conservation. Since the first report of the induction of somatic embryogenesis in Daucus carota (Steward & al. 1958), this technique of micropropagation has been successfully applied to the propagation of a great number of angiosperms and gymnosperms, including some genera of the Iberian Apiaceae endemic in Portugal (Castroviejo, S. & al. 2003), such as Bunium persicum (Grewal, 1996), Thapsia garganica (Jager & al. 1993), Eryngium foetidum (Ignacimuthu & al. 1999) and Angelica sinensis (Tsay & Huang 1998). The objective of the present study was to establish reliable protocols for the propagation of Angelica pachycarpa Lange that can be further used for essential oil production and conservation. MATERIAL AND METHODS Mature seeds of Angelica pachycarpa were collected in the field (Berlenga Islands) and kept at room temperature until the initiation of the experiments. Seeds were soaked in ethanol solution (90% v/v) for 1 min., surface sterilized in 7% (v/v) hypochlorite solution for 20 min. and left in sterilized water overnight. Following three washes in sterile double-distilled water the seeds were germinated in test tubes containing MS (Murashige & Skoog 1962) medium reduced at half strength and 0.087M sucrose. Cultures were kept in a greenhouse and submitted to a daily illumination regime of 15-20 μmol m -2 s -1 photosynthetically active radiation provided by cool-white fluorescent lamps. For shoot proliferation shoot tips (5 mm) from 4-6 week- old seedlings were cultured on MS medium supplemented with 0.44, 1.76 and 4.4 μM BA (benzyladenine) and 0.087 M sucrose. All experiments were carried out three times. For each treatment a minimum of 13 and a maximum of 40 explants were used. The number of shoots formed from shoot tips was evaluated after 4 weeks of culture and the initial explant inoculated for another 4 weeks in the same culture conditions. Roots obtained from the same seedlings were cut into segments of 0.5-1.0 cm long and cultured in the same basal medium containing 4.5 μM 2,4-D (2,4- dichlorophenoxyacetic acid) to induce somatic embryo formation. Root segments from in vitro rooted plantlets were also used for the same purpose. All media were