Abstract
Despite the potential of cellular RNA as a tool for the study of bacterial activities in natural environments, cellular RNA has rarely been measured because of methodological limitations. In a previous study, we quantitatively analyzed single‐cell RNA content with multiple 16S rRNA‐targeted fluorescent probes. In the present study, we explore the potential of this approach with 12 samples of natural planktonic bacteria collected from coastal water over a 6‐month period (late winter to summer). We measured fluorescence from single cells hybridized with multiple probes, cellular DNA and RNA by ethidium bromide (EtBr) fluorometry, and [3H]thymidine (TdR) incorporation rates. Fluorescence of the probe‐hybridized cells was highly correlated to cellular RNA determined by EtBr fluorometry. Winter and summer samples showed different patterns of fluorescence‐frequency distribution. Temperature had a large positive effect on TdR incorporation rates, but there was a negative correlation with cellular RNA content; only the samples taken at elevated summer temperatures showed a positive relationship between TdR incorporation and cellular RNA content. We estimated that the RNA content of the cells not visibly labeled with five probes was <0.3 fg.
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