Pulsed-field gel electrophoresis analysis of virus assemblages present in a hypersaline environment.

A method for analyzing virus assemblages in aquatic environments was developed and used for studying the highest-salinity ponds (from 13.4 to 35% salinity) from a multi-pond solar saltern in Alicante, Spain. The protocol consisted of a series of concentration and purification steps including tangential flow filtration and ultracentrifugation, followed by the preparation of total viral nucleic acids that were subsequently separated by pulsed-field gel electrophoresis. For every sample analyzed, a characteristic DNA pattern was obtained, whose complexity was related to viral diversity. The comparison of our results with a similar analysis carried out with marine virus assemblages shows that, as expected, the viral diversity corresponding to the analyzed hypersaline environment is considerably lower than that of a marine environment.

[1]  C. Cantor,et al.  Pulsed-field gel electrophoresis and the technology of large DNA molecules. , 1988 .

[2]  G. Bratbak,et al.  High abundance of viruses found in aquatic environments , 1989, Nature.

[3]  G. Bratbak,et al.  Enumeration and biomass estimation of planktonic bacteria and viruses by transmission electron microscopy , 1990, Applied and environmental microbiology.

[4]  Jed A. Fuhrman,et al.  Viral mortality of marine bacteria and cyanobacteria , 1990, Nature.

[5]  A. Klieve,et al.  Estimation of ruminal bacteriophage numbers by pulsed-field gel electrophoresis and laser densitometry , 1993, Applied and environmental microbiology.

[6]  F. Rodríguez-Valera,et al.  Evaluation of prokaryotic diversity by restrictase digestion of 16S rDNA directly amplified from hypersaline environments , 1995 .

[7]  F. Rodríguez-Valera,et al.  Sequencing of Bacterial and Archaeal 16S rRNA Genes Directly Amplified from a Hypersaline Environment , 1995 .

[8]  A. Monaco.  Pulsed field gel electrophoresis: a practical approach. , 1995 .

[9]  J. Nolan,et al.  Natural variability and diurnal fluctuations within the bacteriophage population of the rumen , 1996, Applied and environmental microbiology.

[10]  G. Bratbak,et al.  Viral lysis and bacterivory as prokaryotic loss factors along a salinity gradient , 1996 .

[11]  L. Proctor.  Advances in the study of marine viruses , 1997, Microscopy research and technique.

[12]  G. Bratbak,et al.  Occurrence of virus-like particles in the Dead Sea , 1997, Extremophiles.

[13]  Analysis of Marine Viral Assemblages , 1999 .

[14]  Microbiology and biogeochemistry of hypersaline environments , 1999 .

[15]  J. Fuhrman.  Marine viruses and their biogeochemical and ecological effects , 1999, Nature.

[16]  J. Ravel,et al.  Population Dynamics of Chesapeake Bay Virioplankton: Total-Community Analysis by Pulsed-Field Gel Electrophoresis , 1999, Applied and Environmental Microbiology.

[17]  J. Rodríguez,et al.  Enumeration and isolation of viral particles from oligotrophic marine environments by tangential flow filtration. , 1999, International microbiology : the official journal of the Spanish Society for Microbiology.

[18]  N. Guixa-Boixereu,et al.  The microbial food web along salinity gradients. , 2000, FEMS microbiology ecology.

[19]  C. R. Bell,et al.  Microbial Biosystems: New Frontiers , 2000 .