Encapsulation of moss buds: an efficient method for the in vitro conservation and regeneration of the endangered moss Splachnum ampullaceum

In vitro culture techniques are usually employed for ex situ conservation of endangered plant species. However, encapsulation to preserve threatened bryophytes is scarcely used, and only as a pretreatment prior to cryopreservation. In our study, two different methods of germplasm conservation, involving calcium-alginate encapsulation of moss material, were assessed. The plant material used was gametophyte buds (gametophores) of Splachnum ampullaceum Hedw., a rare species of moss. Moss regeneration was evaluated at different periods of time to examine the efficacy of the technique for moss germplasm conservation. The effects of encapsulation and cold storage on developmental parameters such as protonematal colony diameter, bud length, and number of buds were also studied. The results suggest that moss encapsulation with no prior treatment may be a suitable method for germplasm conservation during long periods of time. With our method survival rates as high as 50% could be reached after 2.5 years of cold storage versus 0% of 24-h cryopreserved beads. This technique together with cold storage, avoiding freezing, may be especially important in desiccation intolerant mosses.