Direct PCR Amplifica tion,Sequ encing

From Melted Ice

Acknowledgments.We thank C. J. K. Wang for providing assistancein the cultural and taxonomic aspects of thisstudy. We also thank H. Fan for technicalassistance and J. D. Castello for reviewing themanuscript.

ReferencesAbyzov, S. S. G. (1993) Micro-organisms in the Antarctic

ice, in Antarctic Microbiology (ed. E. 1. Friedman), pp.265-295. New York: John Wiley & Sons, Inc.

Bartholomew, J. W. & Paik, G. (1966) Isolation andidentificationofobligatethermophilicsporeformingbacillifrom oceanbasin cores.Journal ofBacteriology92:635-638.

Catranis, C. & Starmer, W. T. (1991) Micro-organismsentrapped in glacial ice. Antarctic Journal of theUnited States 26: 234-236.

Cross, T. & Atwell, R. W. (1974) Recovery of viablethermoactinomycete endospores from deep mudcores, in Spore Research (eds. A. N. Barker, G. W.Gould & J. Wold), pp. 11-20.London: Academic Press.

Gest,H.&Mandelstam, J. (1987)Longevity ofmicro-organismsin natural environments. Microbiological Sciences4:69-71.

Lindahl, T. (1993) Instability and decay of the primarystructure of DNA. Nature 362: 709-715.

Liu, Y., Ammirati, J. F. & Rogers, S. O. (1997)Phylogenetic Relationships of Dermocybe andCortinarius species based on nuclear ribosomal DNAinternal transcribed spacers. Canadian Journal ofBotany. 75: 519-532.

Ma,1. J.,Fan,H.,Catranis, C.,Rogers,S. 0., & Starmer, W.T.(1997) Isolation and characterization of fungi entrappedin glacial ice. Inoculum 48(3):MSA Abstracts, p.23.

Reeh, N., Johnsen, S. J. & Dahl-Jensen, D. (1985) Datingthe Dye 3 deep ice core by flow model calculations, inGreenland Ice Core: Geophysics, geochemistry, and theEnvironment, Geophysical Monograph 33 (eds. C. C.Langway, Jr., H. Oeschger & W. Dansgaard), pp. 57-65. Washington, D. C.: American Geophysical Union.

Rogers, S. O. & Bendich, A. J. (1985) Extraction of DNAfrom milligram amounts of fresh, herbarium andmummified plant tissues.Plant MolecularBiology 5:69-76.

Seaward, A. C., Cross, T. & Unsworth, B. A. (1976)Viablebacterial spores recovered from an archaeologicalexcavation. Nature 261: 407-408.

Shi, T., Reeves, R. H., Gilichinsky, D. A. & Friedman, E.1. (1997) Characterization of viable bacteria fromSiberian permafrost by 16S rDNA sequencing.Microbial Ecology 33: 169-179.

Volume 13, Part 2, May 1999

Ice Core From Stor age Facility

I 11111/1/

~ III I \ \ \'

~~~~~Section Selected Surface Trea ted with Inner Core Separated

UV Irradiation (middle From Outer Shellcovered with Aseptica lly @ _20°C

aluminum foil) /

lJ--- Filtered Su bsa mpleUnfi ltered .. (taken through same processes asSuh sample with unfiltered samples below)

~ ~Inn er Core Melted

@ 22°C(Stored @ .20°C)

]BACTERIA AND FUNGUS ASSAYS

VIRUS ASSAYS / \

Cultures

/8 Media

(with replicates)

+VariousIncuhationTimes and

Temp eratures

Micros~ 1~ Extraction,Examinations PCR Amplilication,

'" Sequencing "-Characterize Sequ ence Compar isons

and/or Identify .. (DNA DatabaseSearches and

Phylogenetic Ana lyses)

Fig 2 Protocol for ice core isolation, culturing, and DNA sequencing.

Sneath, P. H. A. (1962) Longevity of micro-organisms.Nature 195: 643-646.

Swofford, D. (1993) PAUP: phylogenetic analysis usingparsimony, Version 3.1.1, University of Illinois,Champaign, 11.

Taylor, K. C., Mayewski, P. A., Alley, R. B., Brook, E. J.,Gow, A. J., Grootes, P. M., Meese, D. A., Saltzman, E.S., Severinghaus, J. P., Twickler, M. S., White, J. W.C., Whitlow, S., & Zielinski, G. A. (1997) TheHolocene-Younger dryas transition recorded atSummit, Greenland. Science 278: 825-827.

Vishniac,H. S. (1996) Biodiversity of yeasts and filamentousmicrofungi in terrestrial Antarctic ecosystems.Biodiversity and Conservation 5:1365-1378.

The BMS HomepageThe following are recent additions and amendments to the BMS Homepage (underlined entries are links):

• Foray Box (References for Identifying Fungi), has been updated to January 1999.

• Library: Books, has been reformatted and updated to 1994by integrating a supplement.

• Below Ground Research at York describes their work on roots and mycorrhizas.

• The Warwickshire Fungus Survey has been added to Fungus Groups.

• Photographs of fungi: The Quoditch Moor Nature Research has a nice selection.

RTM

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