ISHS Acta Horticulturae 812: III International Symposium on Acclimatization and Establishment of Micropropagated Plants
MICROPROPAGATION OF CISTUS CLUSII DUNAL: A VERY RARE SPECIES IN ITALY |
| Authors: | C. Ruta, C. Ferraro, I. Morone Fortunato |
| Keywords: | endangered species, in vitro propagation, germplasm conservation, disinfection, 6-benzylaminopurine (BAP), indole butiric acid (IBA) |
| Abstract:
Cistus clusii Dunal (Cistaceae) is a spontaneous shrub of the Mediterranean flora, which is very rare in Italy and critically endangered in the Apulia region. This species, in fact, has a very restricted distribution and in Apulia it only occurs in the small area of Gargano National Park near Lesina between the mouth of the Fortore River and the Mileto Cape. Micropropagation can provide a rapid method for the multiplication of endangered species. The objective of this study was to define a rapid system to multiply C. clusii shoots in vitro to increase the plant number for germplasm conservation. Shoot tips and nodes from field-grown plants were cut and surface-sterilized in 70% (v:v) ethanol for 2 min. and then immersed in 20% (v:v) commercial hypochlorite solution for 20 min. or in a 0.05% HgCl2 solution for 15 or 20 min. and then washed with sterile distilled water. The explants were cultured on a medium containing macronutrients (Murashige and Skoog, 1962) and micronutrients (Nitsch and Nitsch, 1972), Fe-EDTA (30 mg L-1), thiamine HCl (0.4 mg L-1), myoinositol (100 mg L-1), sucrose (30 g L-1) and agar (7 g L-1). Plant growth regulators were incorporated in the medium using varying concentrations and combinations to induce proliferation and rooting. All explants were incubated at 23°C under a light intensity of 60 μEs-1 m-2 with 16 h light/8 h dark photoperiod. After initial problems with disinfection were solved, the explants rapidly formed axillary buds on culture medium containing 6-benzylaminopurine (BAP 0.5 mg L-1) while the best response for rooting was obtained with a 0.1 mg L-1 indol butiric acid (IBA) supplement. Rooted plantlets were acclimatized to greenhouse conditions. The success of this work indicates that micropropagation can be a useful tool for the conservation of Cistus clusii Dunal. | |
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