Abstract

Genome segmentation is mainly thought to facilitate reassortment. Here, we show that segmentation can also allow differences in segment abundance in populations of bluetongue virus (BTV). BTV has a genome consisting in 10 segments, and its cycle primarily involves periodic alternation between ruminants and Culicoides biting midges. We have developed a reverse transcription-quantitative PCR (RT-qPCR) approach to quantify each segment in wild BTV populations sampled in both ruminants and midges during an epizootic. Segment frequencies deviated from equimolarity in all hosts. Interestingly, segment frequencies were reproducible and distinct between ruminants and biting midges. Beyond a putative regulatory role in virus expression, this phenomenon could lead to different evolution rates between segments.IMPORTANCE The variation in viral gene frequencies remains a largely unexplored aspect of within-host genetics. This phenomenon is often considered to be specific to multipartite viruses. Multipartite viruses have segmented genomes, but in contrast to segmented viruses, their segments are each encapsidated alone in a virion. A main hypothesis explaining the evolution of multipartism is that, compared to segmented viruses, it facilitates the regulation of segment abundancy, and the genes the segments carry, within a host. These differences in gene frequencies could allow for expression regulation. Here, we show that wild populations of a segmented virus, bluetongue virus (BTV), also present unequal segment frequencies. BTV cycles between ruminants and Culicoides biting midges. As expected from a role in expression regulation, segment frequencies tended to show specific values that differed between ruminants and midges. Our results expand previous knowledge on gene frequency variation and call for studies on its role and conservation beyond multipartite viruses.

Highlights

  • ACIRAD, UMR ASTRE, Montpellier, France bASTRE, CIRAD, INRA, Univ Montpellier, Montpellier, France cUMR1161 Virologie, ANSES, INRA, Ecole Nationale Vétérinaire d’Alfort, Université Paris-Est, Maisons-Alfort, France dIstituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise G

  • We developed a SYBR green quantitative PCR approach to quantify genome segments similar to those used with multipartite viruses [1, 3, 4]

  • The SYBR green assay was chosen over other quantitative PCR (qPCR) assays due to its accuracy, simplicity, and cost efficiency in schemes requiring the validation of many primer pairs in parallel and the analysis of large sample numbers

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Summary

Introduction

ACIRAD, UMR ASTRE, Montpellier, France bASTRE, CIRAD, INRA, Univ Montpellier, Montpellier, France cUMR1161 Virologie, ANSES, INRA, Ecole Nationale Vétérinaire d’Alfort, Université Paris-Est, Maisons-Alfort, France dIstituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise G. IMPORTANCE The variation in viral gene frequencies remains a largely unexplored aspect of within-host genetics This phenomenon is often considered to be specific to multipartite viruses. A main hypothesis explaining the evolution of multipartism is that, compared to segmented viruses, it facilitates the regulation of segment abundancy, and the genes the segments carry, within a host. These differences in gene frequencies could allow for expression regulation. Multipartite viruses are a diverse group of viruses, including DNA and RNA viruses, that infect mainly plants [2] Their genomes are composed of several segments, and in contrast to segmented viruses, each segment is encapsidated individually. ASheep, Ovis aries; cow, Bos taurus; midge, Culicoides imicola; Curated, samples in which RT-qPCRs of all segments provided a CT value below 31

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