Aquatic Sciences Meeting, Albuquerque 2001

SS04 Environmental Microbial Genomics (Environmental and Disciplinary Connections)

Date: Wednesday, February 14, 2001, Time: 3:45:00 PM

Location: Brazos

Bertilsson, S, , MIT, Civil and Environmental Engineering, Cambridge, USA, stebe@mit.edu

Polz, M, F, MIT, Civil and Environmental Engineering, Cambridge, USA, mpolz@mit.edu

GENERATION AND APPLICATION OF A FUNCTIONAL DIVERSITY ARRAY FOR AQUATIC MICROORGANISMS

The simultaneous identification of biogeochemical-ecological function and phylogeneitic affiliation of microbial populations is one of the biggest challenges in microbial ecology and aquatic sciences. Here, we present a novel culture-independent approach for rapidly identifying individual populations that actively utilize defined organic substrates by simultaneously screening an entire microbial community. The approach combines a 16S rDNA-based oligonucleotide array (Functional Diversity Array) with full water incubations with various radiotracer substrates. Total RNA extracted from the incubations is hybridized to the community array of species-specific probes allowing the identification of populations that were active in the metabolism of the labeled substrates. Making the array is greatly facilitated by a new technique, presented here, that allows the PCR-generation of species-specific oligonucleotide probes without prior knowledge of the sequence of the target organism. This overcomes both time-consuming design of oligonucleotide probes based on sequence generation and high costs associated with determining hundreds to thousands of 16S rDNA sequences from a microbial community. Thus, the Functional Diversity Array presents an effective way to monitor parameters of microbial populations from growth rate to metabolism of specific substrates.

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Aquatic Sciences Meeting, Albuquerque 2001

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