Collecting Times and Sterilization Methods Affect Tissue Culture of Rare and Endangered Species from Western Alps

Climate change and human actions are compromising the conservation status of natural habitats, spontaneous plants, and animal species. To cope with these changes, the European Union has set up the Natura 2000 Network, a network of sites of community interest and special protection areas created for the protection and conservation of habitats, animal and plant species biodiversity. Among the methods used to conserve biodiversity, micropropagation is an in vitro culture method of plant tissues. The aim of this study was to examine the effect of different collecting times and sterilization methods with the potential to enhance in vitro performance for ex situ conservation of six rare and endangered plant species of the Ligurian and Maritime Alps (Clematis alpina (L.) Miller, Dracocephalum ruyschiana L., Gentiana asclepiadea L., Hyssopus officinalis L., Phyteuma cordatum Balb. and Ruscus hypoglossum L.). Results showed that for both C. alpina and D. ruyschiana, as later in the summer were collected explants as higher values in established explants with shoot induction. While the use of plant preservative mixture in tissue culture media was not effective in increasing explant establishment in the studied species. The findings raise concerns regarding the most promosing in vitro protocols for the multiplication of rare and endangered alpine species.