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Brewer, W. T.

Publications and source records attributed to Brewer, W. T..

2 recordsLinked to original sources

Untangling an insect virome from its endogenous viral elements

AO_SCPLOWBSTRACTC_SCPLOWInsects are an important reservoir of viral biodiversity, but the vast majority of viruses associated with insects have not been discovered. Recent studies have therefore employed high-throughput sequencing of RNA, which has led to rapid advances in our understanding of insect viral diversity. However, insect genomes frequently contain transcribed endogenous viral elements with significant homology to exogenous viruses, complicating the use of RNAseq for viral discovery. In this study, we use a multi-pronged sequencing approach to study the virome of an important agricultural pest and prolific vector of plant pathogens, the potato aphid Macrosiphum euphorbiae. We first used rRNA-reduced RNAseq to characterize the bacteria and viruses found in individual insects. We then characterized the frequency of a heritable Flavivirus and an Ambidensovirus in our population. We next generated a quality draft genome assembly for M. euphorbiae using Illumina-corrected Nanopore sequencing. This analysis showed that the Ambidensovirus, previously described from an RNAseq viral screen, is not a exogenous virus and instead is a transcribed endogenous viral element in the M. euphorbiae genome. Our study generates key insight into an important agricultural pest and highlights a widespread challenge for the study of viral diversity using RNAseq.

genomics↗

Second Harmonic Generation Spectroscopy of Membrane Probe Dynamics in Gram-Positive Bacteria

Bacterial membranes are complex mixtures with dispersity that is dynamic over scales of both space and time. In order to capture adsorption onto and transport within these mixtures, we conduct simultaneous second harmonic generation (SHG) and two photon fluorescence measurements on two different gram-positive bacterial species as the cells uptake membrane-specific probe molecules. Our results show that SHG can not only monitor the movement of small molecules across membrane leaflets, but is also sensitive to higher-level ordering of the molecules within the membrane. Further, we show that the membranes of Staphylococcus aureus remain more dynamic after longer times at room temperature in comparison to Enterococcus faecalis. Our findings provide insight into the variability of activities seen between structurally similar molecules in gram-positive bacteria while also demonstrating the power of SHG to examine these dynamics.\n\nSTATEMENT OF SIGNIFICANCEBacterial membranes are highly adept at discerning and modifying their interactions with different small molecules in their environment. Here we show how second harmonic generation (SHG) spectroscopy can track the dynamics of structurally similar membrane probes in two gram-positive bacterial species. Our results reveal behavior that is dependent on both the probe molecule and the membrane composition. Specifically, we observe flip-flop between leaflets for one molecule, while the other molecule produces a signal indicative of larger scale ordering in the membrane. These phenomena can all be explained by considering potential differences in the membrane fluidity and surface charge between the two bacterial species. Overall, our work highlights the dynamic differences between bacterial membranes and SHGs sensitivity to probing these systems.

biophysics↗