Isolating "Uncultivable" Microorganisms in Pure Culture in a Simulated Natural Environment

Tiedje J. M., ASM News 60, 524 (1994).

D. Allsopp R. R. Colwell D. L. Hawksworth Microbial Diversity and Ecosystem Function (CAB International Wallingford UK 1995).

J. T. Staley M. P. Bryant N. Pfennig J. G. Holt Eds. Bergey's Manual of Systematic Bacteriology (Williams & Wilkins Baltimore MD 1989).

Young P., ASM News 63, 417 (1997).

10.1016/S0065-2911(08)60166-6

S. G. Giovannoni M. Rappé in Microbial Ecology of the Oceans D. L. Kirchman Ed. (Wiley-Liss New York 2000) pp. 47–84.

10.1038/345060a0

10.1128/aem.56.3.782-787.1990

10.1038/345063a0

10.1073/pnas.89.12.5685

10.1038/356148a0

10.1073/pnas.91.5.1609

10.1128/AEM.66.4.1617-1621.2000

Butkevich V. S., Trans. Oceanogr. Inst. Moscow 2, 5 (1932).

10.4319/lo.1959.4.2.0128

C. E. ZoBell Marine Microbiology: A Monograph on Hydrobacteriology (Chronica Botanica Waltham MA 1946).

10.1128/aem.36.3.506-512.1978

R. R. Colwell D. J. Grimes in Nonculturable Microorganisms in the Environment R. R. Colwell D. J. Grimes Eds. (American Society for Microbiology Washington DC 2000) pp. 1–6.

10.1146/annurev.mi.39.100185.001541

Details of experimental procedures are available on Science Online at www.sciencemag.org/cgi/content/full/296/5570/1127/DC1.

10.1007/s002489900049

10.1128/AEM.64.7.2691-2696.1998

10.1128/AEM.66.4.1715-1719.2000

L. S. Clesceri A. E. Greenberg A. D. Eaton Standard Methods for the Examination of Water and Wastewater (American Public Health Association Washington DC ed. 20 1998).

10.1139/m70-087

D. R. Boone R. W. Castenholz G. Garrity Eds. Bergey's Manual of Systematic Bacteriology (Springer-Verlag Berlin ed. 2 2001).

10.1099/00207713-44-4-846

D. J. Grimes A. L. Mills K. H. Nealson in Nonculturable Microorganisms in the Environment R. R. Colwell D. J. Grimes Eds. (American Society for Microbiology Washington DC 2000) pp. 209–227.

Out of 60 chamber–to–petri dish passages 19 were successful.

Out of 34 petri dish–to–petri dish passages 33 were negative and one resulted in the mixed culture MSC1/MSC2 (Fig. 3 D and E).

Out of 34 petri dish–to–chamber passages 27 were successful.

The detailed sequence comparison was difficult because of the abundant unspecified sites in the A. nitrofigilis sequence available in GenBank. The next closest relative of MSC2 is an uncultured Arcobacter [96% 16 S rRNA similarity (20)].

F. C. Tenover C. L. Fennel in Prokaryotes A. Ballows H. G. Trüper M. Dworkin W. Harder K.-H. Schleifer Eds. (Springer-Verlag Berlin 1992) pp. 3488–3511.

G. M. Dunny S. C. Winans Eds. Cell-Cell Signaling in Bacteria (American Society for Microbiology Washington DC 1999).

We thank W. Fowle and D. Woebken (Northeastern University Electron Microscopy Center) for help with scanning electron microscopy. We also thank E. Jarroll and M. Kaeberlein for comments on earlier versions of the paper. This work was funded by the U.S. National Science Foundation (grant OCE-0102248).