Molecular Analysis of theIn SituGrowth Rates of Subsurface Geobacter Species
ABSTRACTMolecular tools that can provide an estimate of thein situgrowth rate ofGeobacterspecies could improve understanding of dissimilatory metal reduction in a diversity of environments. Whole-genome microarray analyses of a subsurface isolate ofGeobacter uraniireducens, grown under a variety of conditions, identified a number of genes that are differentially expressed at different specific growth rates. Expression of two genes encoding ribosomal proteins,rpsCandrplL, was further evaluated with quantitative reverse transcription-PCR (qRT-PCR) in cells with doubling times ranging from 6.56 h to 89.28 h. Transcript abundance ofrpsCcorrelated best (r2= 0.90) with specific growth rates. Therefore, expression patterns ofrpsCwere used to estimate specific growth rates ofGeobacterspecies during anin situuranium bioremediation field experiment in which acetate was added to the groundwater to promote dissimilatory metal reduction. Initially, increased availability of acetate in the groundwater resulted in higher expression ofGeobacter rpsC, and the increase in the number ofGeobactercells estimated with fluorescentin situhybridization compared well with specific growth rates estimated from levels ofin situ rpsCexpression. However, in later phases, cell number increases were substantially lower than predicted fromrpsCtranscript abundance. This change coincided with a bloom of protozoa and increased attachment ofGeobacterspecies to solid phases. These results suggest that monitoringrpsCexpression may better reflect the actual rate thatGeobacterspecies are metabolizing and growing duringin situuranium bioremediation than changes in cell abundance.