In-vitro setting and multiplication of Pulicaria microcephala, an endemic species from Berlengas Island
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1
MARE – Marine and Environmental Sciences Centre, ESTM, Instituto Politécnico de Leiria, Portugal
The Berlengas archipelago is located in the Atlantic Ocean, on the Portuguese continental shelf, on the western side of Iberian Peninsula, close to Cape Carvoeiro (Peniche). It distances approximately 5.7 miles from mainland. It is a protected area since 1981. In 2011 Berlengas’ became part of the list of patrimony of UNESCO, aiming to conserve representative terrestrial and marine ecosystems of the Portuguese coast.
As a result to insular isolation, and to very difficult climatic and edaphic conditions, terrestrial flora evolved differently from the species of mainland. In fact, these plants struggle to survive with extreme dryness, very little soil, strong winds and large amounts of salt. These harsh conditions lead to speciation process that differentiated three endemic species, of great conservation value: Armeria berlengensis, Herniaria lusitanica subsp. berlengiana and Pulicaria microcephala. P. microcephala, as well as A. berlengensis, is registered in Annex II of the Habitats Directive, due to its conservation relevance, and is considered vulnerable.
The aim of this project, therefore, was to multiply, in vitro, the species P. microcephala. Plant shoots were collected in the autumn 2015 from a natural wild population in the Berlengas Island. Shoots were obtained from different mother plants in similar maturation stages, trying to interfere the least with the natural propagation process in the protected area. The material was sterilized by soaking in a bleach solution, and after that with ethanol. Shoots were then rinsed four times in sterile distilled water and placed initially on culture tubes with Murashige and Skoog (MS) mineral formulation (Sigma) supplemented with 0.3% (w/v) sucrose and 0.2% (w/v) gelrite (Sigma, Germany). After the end of the first week, the contaminated culture tubes were discarded. Culture tubes were maintained in a growth chamber with a 16h-light photoperiod under cool, white fluorescent lamps (60 µmol.m−2.s−1), at a temperature of 23ºC. Shoots were subcultured in fresh medium every 4 weeks. For the assessment of the multiplication rates, shoots with 2-4 cm high were selected as explants, from stocks maintained in establishment medium. The medium used at this stage contained MS mineral salts, 3% (w/v) sucrose, 1ml.L-1 Gamborg vitamin formulation (Sigma), 1.5% (w/v) Phytoagar and different growth regulators. Plant growth regulators tested included various concentrations of cytokinins (BA, KIN), singly or in combination with auxin (NAA, IAA, IBA). The same basal medium was also used as control, without the addition of any growth regulators. This constitutes the basis for the establishment of the full process of micropropagation of this species. The establishment of adequate micropropagation methods may constitute a valuable route for the conservation of this threatened species, and contribute in a near future to the recovery of this species in its natural habitats.
Acknowledgements
This study is being financed by the project LIFE Berlengas - Conserving threatened habitats and species in Berlengas SPA through sustainable management, LIFE13 NAT/PT/000458. This study had the support of Fundação para a Ciência e Tecnologia (FCT), through the strategic project UID/MAR/04292/2013 granted to MARE.
Keywords:
Berlengas,
Micropropagation,
Shoot multiplication,
Endemic species,
Pulicaria microcephala
Conference:
IMMR | International Meeting on Marine Research 2016, Peniche, Portugal, 14 Jul - 15 Jul, 2016.
Presentation Type:
Poster presentation
Topic:
Biodiversity, Conservation and Coastal Management
Citation:
Franco
I,
Mouga
TM and
Afonso
CN
(2016). In-vitro setting and multiplication of Pulicaria microcephala, an endemic species from Berlengas Island.
Front. Mar. Sci.
Conference Abstract:
IMMR | International Meeting on Marine Research 2016.
doi: 10.3389/conf.FMARS.2016.04.00076
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Received:
28 Apr 2016;
Published Online:
13 Jul 2016.
*
Correspondence:
Prof. Clélia N Afonso, MARE – Marine and Environmental Sciences Centre, ESTM, Instituto Politécnico de Leiria, Peniche, 2520-641, Portugal, clelia@ipleiria.pt