RiboSyn is a new method for measuring specific growth rates of distinctmicrobial populations in natural systems

摘要

Measurements of microbial activities are important in understanding thecycling and transformations of organic and chemical compounds in aquatic andterrestrial environments. As such, a number of molecular biology based methodshave been developed to measure microbial diversity, community activities andmicrobial transformations in cultured and mixed-populations. Although thesetechniques provide valuable information about the community composition andnutrient uptake, there are currently no methods that directly measure specific growthrates of distinct microbial populations in environmental samples. Recently, RiboSynwas developed to measure the specific growth rates of distinct microbial populationsby determining the specific rate of ribosome synthesis. In this technique, cells areexposed to chloramphenicol, inhibiting the processing of pre16S rRNA to 16S rRNA.Using reverse transcription and primer extension, the ratio of pre16S rRNA to 16SrRNA can be measured and ribosome doubling time determined. Due to RTRiboSyn'sability to target specific nucleotide sequences, the specific growth rate ofdistinct microbial populations in mixed cultures can be determined. In order tocharacterize community structure and function of specific bacterial populations inmixed natural assemblages, the RiboSyn method can be used parallel with fluorescentin situ hybridization (FISH), to identify and measure growth rates of individualpopulations of bacteria. In this proceeding, we will describe our research objectiveswhich will be presented at the EWRI conference. The ability to measure specificgrowth rates of distinct microbial populations in a sample from a natural systemwould provide greater understanding of the microbial processes responsible for theutilization or transformation of chemical contaminants in the environment.