Autoradiographic method for isolation of diverse microbial species with unique catabolic traits.

Applied and Environmental Microbiology
J DunbarLarry J Forney

Abstract

A novel autoradiographic method for isolation of bacteria with unique catabolic traits was developed to overcome many of the limitations of traditional selective enrichment techniques. The method consists of five steps. (i) An environmental sample is directly plated (without enrichment) on a microporous filter atop a solid medium that allows cultivation of diverse kinds of microorganisms. (ii) Once colonies form, two replicas of the filter are prepared and the colonies are regrown. (iii) The replica filters are starved 24 to 72 h to deplete intracellular carbon reserves and then (iv) placed on Na(inf2)(sup35)SO(inf4)-containing solid media with and without a test compound. (v) Following an incubation period, the replica filters are exposed to film in order to identify colonies that incorporate more (sup35)S into cell biomass in the presence of the test compound than in its absence, providing presumptive evidence for metabolism of the compound. The colonies identified in this manner can be recovered from the master filter. To demonstrate this technique, bacteria capable of degrading benzoate were isolated from a single soil slurry by traditional enrichment as well as by autoradiography. From the enrichment culture, a single isol...Continue Reading

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Citations

Sep 10, 2005·Applied and Environmental Microbiology·Leen De GelderEva M Top
Jan 29, 2021·Water Environment Research : a Research Publication of the Water Environment Federation·Marwa GuembriHadda Imene Ouzari
Oct 20, 2005·Journal of Bioscience and Bioengineering·K Watanabe, P W Baker
Apr 1, 2000·Applied and Environmental Microbiology·J A KlappenbachT M Schmidt
Apr 1, 1997·Applied and Environmental Microbiology·J DunbarL Forney

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