Abstract

Although bacterial host colonization is a dynamic process that requires population growth, studies often focus on comparing bacterial populations at a given time point. However, this may not reflect the dynamics of the colonization process. Time-course assays provide important insights into the dynamics of host colonization but are laborious and may still lack resolution for immediate processes affecting populations. An alternative way to address this issue, using widely accessible tools (such as quantitative PCR [qPCR]), is to take advantage of the relationship between bacterial chromosomal replication and cell division to determine population growth status at the sampling time. Conceptually, the ratio between the number of copies at the origin of replication and that at the terminus of replication should be correlated with the measured bacterial growth rate. This peak-to-trough ratio (PTR) to estimate instantaneous population growth status was tested with the slow-growing plant-pathogenic bacterium Xylella fastidiosa. We found no correlation between PTR and the measured growth rate when using genome-level data but overall sequencing depth of coverage trends matched theoretical expectations. On the other hand, the population growth status of X. fastidiosa was predicted by PTR when using qPCR data, which was improved by the pretreatment of cells with a photoreactive DNA-binding dye. Our results suggest that PTR could be used to determine X. fastidiosa growth status both in planta and in insect vectors. We expect PTR will perform better with fast-growing bacterial pathogens, potentially becoming a powerful tool for easily and quickly assessing population growth status.

Highlights

  • Bacterial host colonization is a dynamic process that requires population growth, studies often focus on comparing bacterial populations at a given time point

  • Our first goal was to determine whether peak-to-trough ratio (PTR) could be used to predict X. fastidiosa growth dynamics at the genome scale, and whether X. fastidiosa sequencing depth of coverage followed expectations based on observations of fast-growing bacteria

  • We showed that quantitative PCR or real-time PCR (qPCR) may provide information on bacterial growth status for a slow-growing plant pathogen using a single time point if two primer pairs are used at opposite regions of the chromosome, one near the origin and another at the terminus of replication

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Summary

Introduction

Bacterial host colonization is a dynamic process that requires population growth, studies often focus on comparing bacterial populations at a given time point. This may not reflect the dynamics of the colonization process. The ratio between the number of copies at the origin of replication and that at the terminus of replication should be correlated with the measured bacterial growth rate. This peak-to-trough ratio (PTR) to estimate instantaneous population growth status was tested with the slow-growing plant-pathogenic bacterium Xylella fastidiosa. We expect PTR will perform better with fast-growing bacterial pathogens, potentially becoming a powerful tool for and quickly assessing population growth status

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