In vitro micropropagation of rare and endangered moss Entosthodon hungaricus (Funariaceae)

Abstract

The moss Entosthodon hungaricus (Boros) Loeske is an European endemic species typical of dry and saline soils extending from the Iberian Peninsula to Aral-Caspian steppes, similarly to some other xerothermic bryophytes. However, the distribution range is fragmented and localities are quite scattered and the species is considered as rare and vulnerable because of its ephemeral characteristics and specialized ecology. With the aim to develop an active protection plan for this species, the ex situ conservation requirements of E. hungaricus were developed. The axenic culture in in vitro conditions were established, and the optimal growth parameters were adjusted to achieve fully developed gametophytes ready to be reintroduced to its native range and other potentially native areas, where this species was once reported but has not been collected in recent times, suggesting its local extinction (i.e. some areas in Vojvodina, N. Serbia). Starting materials were derived from recent herbarium specimens and from fresh materials collected from Hungarian populations. Several means for sterilization of stating material and growing nutritive media were assayed in different regimes of light and temperature. Here we describe the conditions to achieve full plant development and for its micropropagation. Such materials are adequate for ex situ conservation purposes and for experimental introductions in native and potentially native areas. The first axenical culture of E. hungaricus was successfully established, and the first in vitro micropropagation of this rare and endangered species was achieved. Our study contributes to the conservation biology as well as for the potential use of this moss species in biotechnological research.