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Biology subjects

Remnant, E. J.

Publications and source records attributed to Remnant, E. J..

7 recordsLinked to original sources

Primary and secondary antiviral RNAi responses throughout Varroa destructor life stages reveal the vertical transmission of viruses

One of the most devastating threats to global honey bee health is the ectoparasitic mite and viral vector Varroa destructor, yet the transmission dynamics of viruses carried by mites are poorly understood. RNA interference (RNAi) is a major antiviral defence mechanism in invertebrates including Varroa, where actively replicating viruses are degraded into virus-derived small interfering RNAs (vsiRNAs). Insects typically produce 20-22 nt vsiRNAs with sense and antisense polarity, however established viral infections in V. destructor lead to the production of 24-nt antisense vsiRNA fragments, which could indicate the presence of secondary siRNA synthesis. To better understand viral infection and transmission dynamics in V. destructor, we conducted small RNA sequencing of male and female mites throughout development, from egg to reproductive stages. Viral community structure was largely driven by developmental stage, with younger and older life stages clustering separately. We identified five viruses that are consistently degraded into antisense 24-nt vsiRNA across all developmental stages, suggesting that these viruses are transmitted vertically and form part of Varroas core virome. This includes the highly diverse Varroa destructor virus 2 (VDV-2), for which we observe eight distinct VDV-2 strains that simultaneously co-infect individual mites throughout development. In contrast, sense and antisense 23-nt vsiRNA fragments are generated in response to the honey bee pathogen, Iflavirus aladeformis (deformed wing virus A, DWV-A) in eggs, but the vsiRNA size profile transitions to 24-nt antisense fragments at later life stages. Our results suggest that once a virus is first acquired by Varroa, a primary 23-nt sense and antisense antiviral response precedes the production of secondary 24-nt antisense vsiRNAs as the infection progresses. We confirm this observation using synthetic dsRNA, which show both primary and secondary siRNA processing, revealing how exogenous dsRNA processing occurs in Varroa. These results show distinct primary and secondary antiviral RNAi responses across V. destructor life stages and demonstrate how vsiRNA profiles can be used to infer virus transmission routes and long-term persistence within vector populations.

genetics↗

Landscape viromics of introduced honeybees and bumblebees reveal distinct environmental and host-specific effects

Understanding how viral communities vary across co-occurring hosts and environments is essential for assessing species-specific viral risks under changing land use and climate. This is particularly relevant for managing introduced bees, which face persistent viral threats themselves, as well as transmitting plant viruses. Here, we compare RNA viromes of the long-established honeybee (Apis mellifera, introduced to Tasmania in 1831) and the more recent invader, the bumblebee (Bombus terrestris, invasive since 1992), across 14 Tasmanian sites - an island still free of the viral vector, Varroa destructor. Using a metatranscriptomic approach on total RNA from whole bees, we identified insect- and plant-associated viruses and inferred phylogenetic patterns of insect viral sharing, divergence, and potential cross-species transmission. We also assessed spatial and environmental drivers of viral composition, diversity, and richness. Geographic longitude, precipitation, temperature, and pasture percentage influenced the total, insect-, and plant-associated viromes of B. terrestris. In contrast, for A. mellifera, only precipitation and temperature were associated with insect and plant viral alpha diversity and community composition. Phylogenetic analyses revealed that Black Queen Cell virus in A. mellifera from Tasmania has diverged from mainland Australian sequences, and two distinct sub-strains of Lake Sinai virus 1 were shared by both bee species. Lake Sinai virus 3 showed evidence of interspecies transmission between A. mellifera and B. terrestris. Notably, this study provides the first detection of Moku virus in Australian bees and globally in bumblebees, suggesting potential interspecies transmission among social Hymenoptera. Overall, our findings demonstrate local viral diversification and reveal that B. terrestris viromes are more strongly shaped by environmental factors than those of A. mellifera, underscoring the importance of monitoring invasive pollinators as reservoirs and vectors of viral emergence.

evolutionary biology↗

Deformed wing virus genotypes A and B do not elicit immunologically different responses in naive honey bee hosts

Deformed wing virus (DWV), in association with Varroa destructor, is currently the leading factor associated with global honey bee deaths. With the exception of Australia, the virus and mite have a near global distribution, making it difficult to separate the effect of one from the other. Over time, the prevalence of the two main DWV genotypes (DWV-A and DWV-B) has changed, leading to the suggestion that the two strains elicit a different immune response by the host, the western honey bee Apis mellifera. Here we use a honey bee population naive to both the mite and the virus to investigate if honey bees show a different immunological response to DWV genotypes. We examined the expression of 19 immune genes by RT-qPCR and comprehensively analysed the small RNA response in honey bees after experimental injection with DWV-A and DWV-B. We found no evidence to indicate that DWV-A and DWV-B elicit a different immune response in honey bees. We found that RNA interference genes are up-regulated during DWV infection and that the small interfering RNA (siRNA) response is proportional to viral loads, yet does not inhibit the virus from accumulating to high loads. We also found that the siRNA response towards DWV was weaker than the response to another honey bee pathogen, Black queen cell virus. This suggests that DWV is comparatively better at evading antiviral host defences. There was no evidence for the production of virus-derived PIWI-RNAs in response to DWV infection. In contrast to previous studies, and in the absence of V. destructor, we found no evidence that DWV has an immunosuppressive effect in honey bees. Overall, our results advance our understanding of the immunological effect DWV elicits in honey bees.

immunology↗

Virus replication in the honey bee parasite, Varroa destructor

Arthropod vectors such as mites and ticks introduce an alternative viral transmission route between their hosts. The ectoparasitic mite Varroa destructor is the leading threat to the health of Western honey bees (Apis mellifera) primarily through its action as a vector of viruses. However, it is unclear whether viruses transmitted by V. destructor actively infect and replicate in mites, which could facilitate increased transmission and select for more virulent strains. To better understand the role of V. destructor as a vector, we took advantage of differences between bee and mite antiviral RNA interference pathways to identify the host specificity of replicating viruses. We used small RNA sequencing of individual V. destructor mites to examine viral small interfering RNA (vsiRNA) profiles of Deformed wing virus genotypes (DWV-A and DWV-B), associated with colony declines, as well as nine other viruses present in our samples. We found active replication of six V. destructor-associated viruses, including a novel virus, Varroa destructor virus 9 (VDV-9), and replication of two honey bee associated viruses, including both DWV-A and -B genotypes, suggesting that mites are biological vectors for important bee pathogens. We show that the antiviral RNAi response can be used define the host range of viruses in host-parasite interactions, such as honey bees and their parasites, enabling a better understanding of the role of a vector in the evolution and spread of honey bee pathogens.

genetics↗

Abundant small RNAs in the reproductive tissues of the honey bee, Apis mellifera, are a plausible mechanism for epigenetic inheritance and parental manipulation of gene expression

Polyandrous social insects such as the honey bee are prime candidates for parental manipulation of gene expression in offspring. Although there is good evidence for parent-of-origin effects in honey bees the epigenetic mechanisms that underlie these effects remain a mystery. Small RNA molecules such as miRNAs, piRNAs and siRNAs play important roles in transgenerational epigenetic inheritance and in the regulation of gene expression during development. Here we present the first characterisation of small RNAs present in honey bee reproductive tissues: ovaries, spermatheca, semen, fertilised and unfertilised eggs, and testes. We show that semen contains fewer piRNAs relative to eggs and ovaries, and that piRNAs and miRNAs which map antisense to genes involved in DNA regulation and developmental processes are differentially expressed between tissues. tRNA fragments are highly abundant in semen and have a similar profile to those seen in semen in other animals. Intriguingly we find abundant piRNAs that target the sex determination locus, suggesting that piRNAs may play a role in honey bee sex determination. We conclude that small RNAs play a fundamental role in honey bee gametogenesis and reproduction and provide a plausible mechanism for parent-of origin-effects on gene expression and reproductive physiology.

genomics↗

DNA methylation is not a driver of gene expression reprogramming in young honey bee workers

Intragenic DNA methylation, also called gene body methylation, is an evolutionarily-conserved epigenetic mechanism in animals and plants. In social insects, gene body methylation is thought to contribute to behavioral plasticity, for example between foragers and nurse workers, by modulating gene expression. However, recent studies have suggested that the majority of DNA methylation is sequence-specific, and therefore cannot act as a flexible mediator between environmental cues and gene expression. To address this paradox, we examined whole-genome methylation patterns in the brains and ovaries of young honey bee workers that had been subjected to divergent social contexts: the presence or absence of the queen. Although these social contexts are known to bring about extreme changes in behavioral and reproductive traits through differential gene expression, we found no significant differences between the methylomes of workers from queenright and queenless colonies. In contrast, thousands of regions were differentially methylated between colonies, and these differences were not associated with differential gene expression in a subset of genes examined. Methylation patterns were highly similar between brain and ovary tissues and only differed in nine regions. These results strongly indicate that DNA methylation is not a driver of differential gene expression between tissues or behavioral morphs. Finally, despite the lack of difference in methylation patterns, queen presence affected the expression of all four DNA methyltransferase genes, suggesting that these enzymes have roles beyond DNA methylation. Therefore, the functional role of DNA methylation in social insect genomes remains an open question.

genomics↗

Vector-mediated viral transmission favours less virulent viruses

While it is well-established that the ectoparasitic mite Varroa destructor is largely responsible for the widely-reported decline of populations of the Western honeybee Apis mellifera, the exact role the mite plays in honeybee health remains unclear. The last few years have seen a surge in studies associating RNA viruses vectored by the mite with the death of honeybee colonies. Varroa facilitates the spread of RNA viruses because it feeds on developing bee brood and transfers haemolymph from bee-to-bee. Such a change in transmission, from horizontal and vertical to vector-based, is predicted to lead to an increase in virulence of RNA viruses, thus potentially providing an explanation for the observed association between Varroa and certain viruses. Here we document the effect of changing the route of transmission of honeybee viruses contained in the haemolymph of honeybee pupae. We find that a change in mode of transmission rapidly increases viral titres of two honeybee viruses, Sacbrood virus (SBV) and Black queen cell virus (BQCV). This increase in viral titre is accompanied by an increase in virulence. In contrast, the virus most often associated with Varroa, Deformed wing virus (DWV), shows a reduction in viral titre in the presence of SBV and BQCV. In addition, DWV does not cause mortality to honeybee pupae in isolation. Most likely a change in mode of transmission due to the arrival of a vector quickly eliminates the most virulent honeybee viruses resulting in an association between Varroa and less virulent viruses such as DWV. Our work therefore provides empirical evidence for an alternative explanation for the widely-observed association between Varroa and DWV.

evolutionary biology↗