Micropropagation and optimisation of in vitro production of the rare and threatened moss Entosthodon pulchellus (Funariaceae)

The establishment of axenic cultures and the propagation of the rare moss Entosthodon pulchellus (H. Philip.) Brugués (Funariaceae), as well as the optimisation of its ex situ growth conditions and rapid biomass production were the goals of this study. The results obtained also provide insights into the developmental biology of this moss in in vitro conditions, particularly regarding basal media contents with or without supplements of selected plant growth regulators or sugars, i.e. its nutritional needs. The procedure for establishing axenic cultures of this rare and threatened moss species is described. The optimisation protocol of the growth and production conditions is elaborated. The spore germinability of dried herbarium samples remained rather high in the tested laboratory conditions. Suitability tests on different media types showed KNOP basal media to be the best fit for the rapid biomass production of both secondary protonemata and leafy gametophores, without the addition of sugars or plant growth regulators. However, sugar, namely sucrose, induced rapid and massive protonemal development and can be used when this developmental stage is needed.