The assessment of DNA from marine otganisms via a modified salting-out protocol

• Autorzy: Ferrara G B , Murgia B. , Parodi A.M. , Valisano L. , Cerrano C. , Palmisano G. , Bavestrello G. , Sara M.
• Języki publikacji: EN

Abstrakty

EN

We developed a rapid, practical and non-toxic salting-out method for the extraction of DNA from marine organisms, and tested it on two representative species of Porifera and Cnidaria, both living in association with symbiotic zooxanthellae. We tested the efficiency of the protocol by comparing the output of the method for fresh tissue, frozen tissue and tissue stored in ethanol. It proved to be effective for extracting DNA in the case of the methods of preservation considered here, and for obtaining quantities of DNA comparable to those obtained via the traditional approach. The DNA from both species was of good quality. The DNA obtained was amplified by PCR using specific primers for the large ribosomal subunit, allowing the identification of the presence of both the host and symbiont genomes.

Słowa kluczowe

EN

Porifera; extraction; sea organism; Symbiodinium; mortality; benthic organism; polymerase chain reaction; DNA; sea invertebrate; Cnidaria

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2006
• Tom: 11
• Numer: 2
• Opis fizyczny: p.155-160,fig.,ref.

Twórcy

autor

Ferrara G B

• Universita di Genova, Viale Benedetto XV, 16132 Genova, Italy

autor

Murgia B.

autor

Parodi A.M.

autor

Valisano L.

autor

Cerrano C.

autor

Palmisano G.

autor

Bavestrello G.

autor

Sara M.

Bibliografia

• 1. Glynn, P.W. Coral reef bleaching in the 1980s and possible connections with global warming. Trends Ecol. Evol. 6 (1991) 175-179.
• 2. Hayes, M.L., Bonaventura, J., Mitchell, T.P., Prosper, J.M., Shinn, E.A., Van Dolah, F. and Barber, R.T. How are climate and marine biological outbreaks functionally linked? Hydrobiologia 460 (2001) 213-220.
• 3. Muller-Parker, G. and D’Elia, C.F. Interactions between corals and their symbiotic algae. In: Life and Death of Coral Reefs (Birkeland, C. Ed.) Chapman & Hall, New York., 1997, 96-113.
• 4. Wakefield, T.S., Farmer, M.A. and Kempf, S.C. Revised description of the fine structure of in situ “zooxanthellae” genus Symbiodinium. Biol. Bull. 199 (2000) 76-84.
• 5. Goulet, T.L. and Coffroth, M.A. Stability of an octocoral-algal symbiosis over time and space. Mar. Ecol. Prog. Ser. 250 (2003) 117-124.
• 6. Rowan, R., Knowlton, N., Baker, A.C. and Jara, J. Landscape ecology of algal symbiont communities explains variation in episodes of coral bleaching. Nature 388 (1997) 265-269.
• 7. Cowen, R. The role of algal Symbiosis in reefs through time. Palaios 3 (1998) 221-227.
• 8. Hoegh-Guldberg, O. Climate change, coral bleaching and the future of the world’s coral reefs. Mar. Freshw. Res. 50 (1999) 839-866.
• 9. Muscatine, L. and Porter, J.W. Reef corals – mutualistic symbioses adapted to nutrient-poor environments. BioScience 27 (1997) 454-460.
• 10. Junghans, H. and Metzlaff, M. A simple and rapid method for preparation of total plant DNA. Biotechnique 8 (1990) 176.
• 11. Coffroth, M.A., Lasker, H.R., Diamond, M.E., Bruenn, J.A. and Bermingham, E. DNA fingerprints of a gorgonian coral: a method for detecting clonal structure in a vegetative species. Mar. Biol. 114 (1992) 317-325.
• 12. Sinha, R.P, Dautz, M. and Hader, D.P. A simple and Efficient Method for the Quantitative Analysis of Thymine Cyanobacteria, Phytoplancton and Macroalgae. Acta Protozool. 40 (2001) 187-195.
• 13. Chen, C.A. and Yu, J.K. Universal primers for amplification of mithochondrial small subunit ribosomal RNA-encoding gene in scleractinian corals. Mar. Biotechnol. 2 (2000) 146-153.
• 14. LaJeunesse, T.C. Investigating the biodiversity, ecology and phylogeny of endosymbiotic dinoflagellates in the genus Symbiodinium using ITS region. In: Search of a “specie” level marker. J. Phycol. 37 (2001) 866-880.
• 15. Santos, S.R., Gutierrez-Riodriguez, C., Lasker, H.R. and Coffroth, M.A. Symbiodinium sp. Association in the gorgonian Pseudopterogorgia elisabethae in the Bahamas: high level of genetic variability and population structure in symbiotic dinoflagellates. Mar. Biol. 143 (2003) 111-120.
• 16. Crabbe, M.J. A novel method for the transport and analysis of genetic material from polyps and zooxanthellae of scleractinians corals. J. Biochem. Biophys. Methods 57 (2003) 171-176.
• 17. Miller, S.A., Dykes, D.D. and Polesky, H.F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acid Res. 16 (1988), 1215.
• 18. Rawlings, L.H. and Donellan, S.C. Phylogeographic analysis of the green phyton, Morelia viridis, reveals criptic diversity. Mol. Phylogen. Evol. 27 (2003) 36-44.
• 19. Underwood, A.J. Techniques of analysis of variance in experimental marine biology and ecology. Annu. Rev. Ocean. Mar. Biol. 19 (1981) 513-605.