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Pre-print Volume - Posters Topic 1: BIODIVERSITY AND CONSERVATION SCIENCE: CONTRIBUTING TO MANAGEMENT 41 st S.I.B.M. CONGRESS Rapallo (GE), 7-11 June 2010 87 Fig. 1 – Sampling locations of Calderón et al. (2006) (paddle) and of the present work (arrow). Località di campionamento di Calderón et al. (2006) (pagaia) e del presente lavoro (freccia). Each sample consisted of a 6-8 cm long fragment collected from the tip of a colony and preserved in 96% ethanol. Sampling was carried out only on those gorgonians that were at least 50 cm high. DNA was extracted from 20 polyps of each fragment using a Salting Out protocol modified from Aljanabi & Martinez (1997). The new protocol employed a highly concentrated lysing solution (EDTA 100 mM; Tris HCl 10 mM pH 7.5; SDS 0.6%; NaCl 400 mM) in which tissue was incubated overnight. DNA was PCR-amplified using the COI primers reported in Calderón et al. (2006). (COI Cni F: 5’-GGYACTYTATATTTACTATTTGG-3’; COI Cni R: 5’- CCSGCAGGATCAAAGAAWGTTG-3’). Amplified products were then purified and sequenced by a commercial company. Two individuals were sequenced from each location and the obtained nucleotide sequences aligned using the software BioEdit vers. 7.0.9.0 (Hall, 1999). Results – PCR amplifications delivered fragments of 545 bp size. Nucleotide sequences were inserted in GenBank and they confirmed amplification of the COI region. All obtained sequences were identical to each other and to the one of P. clavata deposited in GenBank by Calderón et al. (2006) (GenBank access n°: AY827539.1). Conclusions – The genetic homogeneity at the COI locus of P. clavata, in individuals sampled at locations that were several hundred kilometers distant, confirms Calderón et al.’s (2006) results even after enlargement of the study area and use of a higher number of individual colonies. The lack of polymorphism at the COI region in P. clavata could be attributed to a slow rate of mutation of the gene or efficient repairing system of this species’ mtDNA. To fully accomplish the objective of this work the characterization of genetic markers with a sufficient degree of polymorphism is needed. References ALJANABI S.M., MARTINEZ I. (1997) – Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucleic Acids Research, 25: 4692-4693. CALDERÓN I., GARRABOU J., AURELLE D. (2006) – Evaluation of the utility of COI and ITS markers as tools for population genetic studies of temperate gorgonians. Journal of Experimental Marine Biology and Ecology, 336: 184-197. CARPINE C., GRASSHOF M. (1975) – Les Gorgonaires de la Méditerranée. Bulletin de l'Institut Océanographique, 71: 1-140. CUPIDO R., COCITO S., BORDONE A., SGORBINI S., SANTANGELO G. (2008) – Response of a gorgonian (Paramuricea clavata) population to mortality events: recovery or loss? Aquatic Conservation: Marine and Freshwater Ecosystems, 18: 984-992. HALL T.A. (1999) – BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41: 95-98.
Topic 2 / Tema 2 Pre-print Volume Topic 2: MARINE ORGANISMS AND ECOSYSTEMS AS MODEL SYSTEMS MARINE ORGANISMS AND ECOSYSTEMS AS MODEL SYSTEMS ORGANISMI MARINI ED ECOSISTEMI QUALI SISTEMI MODELLO 41 st S.I.B.M. CONGRESS Rapallo (GE), 7-11 June 2010 88
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Castellano M.......................
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