C1 In vitro production of propagation material for reinforcement and reintroduction

In order to carry out the activities planned in actions C2, C3 and C7, the target species will be propagated through alternative techniques in respect to the classic multiplication methods (e.g. multiplication by seed, cuttings) using new in vitro propagation protocols. In recent years, many initiatives have promoted the potential of biotechnologies in the field of ex situ conservation of plant biodiversity. In particular, among the various applications of in vitro cultures, micropropagation plays an important role and so far has provided extremely interesting results. In fact, micropropagation allows to obtain in a short time an exponential number of specimens starting from a minimum stock of plant material gathered in the field, without involving significant changes to the conservation status of the natural populations. Moreover, this method is particularly suitable for the multiplication of the target species, especially considering the problems of genetic pollution which C. maritima is currently exposed to, due to the phenomena of hybridization with C. fulgida. In fact, the plant material to be used for multiplication will be taken from pure genetic lineages, after their identification through action A1.

During the first phase the material collected in the field will be sterilized and introduced in vitro in order to produce a first generation of individuals to be used during the successive phases of multiplication. Then, in vitro multiplication will be performed starting from nodal explants obtained in the previous phase, placed in the best growing conditions, as already identified from previous studies made at the IBBR laboratories of Palermo. The individuals obtained in this phase will be used in the subsequent rooting and acclimatization phases, also using already validated protocols. In particular, the rooting of the micro-cuttings will be carried out using the most effective hormones for the production of a root system functional to the transfer in open field. After the induction phase of the root system, the new plants will be gradually acclimatized until they are fully adapted to the open field conditions (see action C6). Finally, before transfer to the field, it will be necessary to evaluate the genetic stability of the new individuals obtained, as described in action D3.

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