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Pannebakker, B. A.

Publications and source records attributed to Pannebakker, B. A..

17 recordsLinked to original sources

Females do not avoid but prefer superparasitism in the parasitoid wasp Melittobia australica

To reduce competition over limited resources, organisms often avoid resources already used by others. In parasitoid insects, females are generally expected to avoid parasitizing hosts already parasitized by other females, called superparasitism, and this prediction has been broadly supported across numerous species. However, in the parasitoid wasp Melittobia australica, multiple females were observed ovipositing on the same host despite the presence of nearby unparasitized hosts in the field. In this study, we first analysed field data using randomization tests, and found that females strongly aggregated on particular hosts rather than distributing randomly across hosts. We then conducted laboratory experiments to examine (1) whether this aggregation reflects an active preference for superparasitism and (2) whether such behaviour could improve parasitism efficiency. A choice experiment, in which females were provided with a parasitized and an unparasitized host, showed that females preferred superparasitism when hosts were non-diapausing. This result indicates that females of M. australica not only engage in superparasitism, but actively prefer parasitized hosts. Another experiment, in which varying numbers of females were allowed to oviposit on a host, found that offspring number per female decreased with increasing female number on diapausing hosts, whereas it remained constant irrespective of female number on non-diapausing hosts. This host-dependent result suggests that multiple females can more effectively paralyze vigorous non-diapausing hosts, thereby offsetting the negative effects of offspring competition. As Melittobia females are likely to attack non-diapausing hosts in most cases in the field, including those in the present study, this benefit of parasitism by multiple females may help explain their preference for superparasitism, in contrast to many other parasitoid species.

animal behavior and cognition↗

Local population genetic structure of Andrena flavipes (Hymenoptera, Andrenidae), a nest-philopatric communal mining bee, reveals isolation-by-distance and high inbreeding

Wild bee populations are declining globally and understanding how species-intrinsic traits like nest-philopatry shape local population genetic structure is increasingly important for their conservation. However, few studies have explored this at local spatial scales. To fill this gap, we used SNPs in the nuclear vitellogenin (Vg) gene to assess the local population structure and genetic diversity of the philopatric yellow-legged mining bee, Andrena flavipes, in the Netherlands, one of the most fragmented and urbanized countries in Europe. We investigated whether its reported nest-philopatry, combined with a fragmented landscape, would lead to genetic differentiation and isolation-by-distance (IBD) between populations, and low genetic diversity and inbreeding within populations. A total of 144 A. flavipes bees were sequenced for the full-length Vg gene and genotyped for SNPs. Despite high overall genetic variation, we found a heterozygote deficit and positive inbreeding coefficient in most populations, suggesting non-random mating. Low genetic differentiation was detected between populations, with no distinct genetic clusters, but with a pattern of IBD. These results suggest extensive but spatially structured gene flow, consistent with philopatric behaviour, and indicate that increasing isolation between nest-aggregations potentially leads to genetic differentiation over time. Our findings demonstrate that even in a common and widespread species, nest-philopatry and landscape fragmentation can drive inbreeding and spatially structured gene flow at local scales, highlighting the importance of maintaining habitat connectivity and semi-natural elements in agricultural and urbanized landscapes to sustain wild bee populations.

genetics↗

Enemies, more than sex, shape butterfly post-mating odor evolution

Not all odors influencing mating behavior evolve as sex pheromones. Female butterflies post-mating odors have been considered species-specific anti-aphrodisiac pheromones shaped by sexual selection but may also serve broader ecological roles shaped by natural selection. Males transfer odors to females that repel rivals, yet the widespread use of these compounds across phyla makes them targets for eavesdropping, such as by phoretic egg parasitoids. We show that in cabbage white butterflies (Pieris spp.), these odors are highly variable and attract parasitoids, deter predators, and influence oviposition. Using gas chromatography and electroantennography, we demonstrate that odor emission and perception lack species-specificity: compounds once thought unique to P. brassicae and P. rapae are shared across Pieridae. In P. napi, odor variation among populations correlates with parasitoid pressure, but not with latitude, genetic distance, or mating frequency, suggesting ecological rather than sexual drivers. In P. brassicae, CRISPR/Cas9 disruption of odor perception alters oviposition and increases susceptibility to parasitism. Moreover, these odors render females unpalatable to birds. Together, our results show that post-mating odors in Pieris butterflies may act as aposematic signals. We provide evidence that these signals evolve under multiple selective pressures, balancing deterrence of mates and predators, parasitoid avoidance, and host-plant interactions. These findings suggest that chemical signals should be viewed as integrating ecological and reproductive pressures, rather than being interpreted solely through the lens of sexual communication.

ecology↗

Bombus terrestris Complementary Sex Determiner (BtCSD) is identified as a conserved hymenopteran sex determination region: evolution, breeding, and conservation implications

All Hymenoptera (bees, sawflies, ants, wasps) are haplodiploid. Haploid males develop from unfertilized eggs and diploid females from fertilized eggs. Many species also have single locus complementary sex determination (sl-CSD): hemizygosity of a CSD gene results in normal males and heterozygosity in normal females, but homozygosity typically yields sterile diploid males. Populations that lose CSD allelic diversity to critical level, increasingly produce detrimental diploid males. Emblematic of this problem are the bumblebees. Bumblebees are one of the fastest declining animal groups worldwide, with CSD diversity loss playing a likely role, especially in isolated populations. Presently utilized and potential future species in agriculture are also challenged by diploid males in production. Population management of both requires exact knowledge of the CSD locus. Here we report the identity of Bombus terrestris CSD, a bumblebee model that is also a major commercial pollinator. As CSD must be reciprocally homozygous in diploid males and heterozygous in diploid females, we whole-genome-sequenced these for four colonies. From this, we identified B. terrestris CSD (BtCSD) on Chromosome 2: a 26 Kb region with a long non-coding RNA region (lncRNA) and a proximate hypervariable region. Comparative analyses indicated conservation of this region across infraorder Aculeata, including other bumblebees, mason bees, and ants. This supports a deep-rooted ancestral origin for BtCSD despite a lack of homology with con-familial honey bee csd and rapid evolutionary divergence of insect sex determination cascades. Our analyses do not support the lncRNA region as functional CSD, as proposed for other taxa, because of inadequate variation to constitute distinct alleles. Rather, a downstream region is implicated for its massive structural variation and candidate coding genes including Creld and two copies of aCOP. We discuss how this identification of BtCSD has critical implications for hymenopteran evolution and bumblebee population management.

genomics↗

Investigating multiple types of resistance against a homing gene drive in European populations of Drosophila melanogaster

Gene drive technology may be a valuable tool for addressing several contemporary challenges, including combating disease vectors, conserving biodiversity, and controlling agricultural pests. Homing gene drives spread through a population by copying themselves onto the homologous chromosome in the germline of heterozygous individuals. However, it is possible that resistance will evolve against homing gene drives, especially if the goal is to suppress or eliminate a pest population so that resistance alleles have a large selective advantage over the gene drive. Resistance can result from a simple mutation at the drives target site, which is found in many studies but can potentially be avoided by improving drive design. However, a more complex polygenic type of resistance could also evolve through selection on standing genetic variation that affects the efficiency of the spread of the gene drive. In this study, we test an efficient homing gene drive in genetically diverse lines of Drosophila melanogaster, collected from across Europe by the DrosEU Consortium. We find that the gene drive shows considerable variability in homing efficiency, but that none of this variability can be ascribed to heritable genetic effects. Selection for complex resistance is thus unlikely and will be inefficient, probably still giving a gene drive enough time to fixate in the population. However, although our tested gene drive targets a highly conserved haploinsufficient gene with two gRNAs, we find simple resistance alleles in viable offspring. Half of these are the product of end-joining repair instead of homing and may still carry heavy fitness costs. However, the other half are the result of partial homing events. These alleles indicate that resistance could likely evolve against this gene drive in a simple, non-polygenic way. Therefore, more effective strategies may be required to address simple resistance mutations, whereas complex resistance may be unlikely to pose a substantial barrier to the employment of at least certain types of gene drive.

genetics↗

Genomic prediction in insects: a case study on wing morphology traits in the jewel wasp Nasonia vitripennis

BackgroundBiological control is a sustainable strategy to combat agricultural pests. Yet, due to legislation, importing non-native biocontrol agents is increasingly restricted. Thus, selective breeding of biocontrol traits of native species is suggested to enhance performance of existing biocontrol agents. Genomic prediction is a new alternative to exploit genetic variation for improving biocontrol efficacy. This study aims to establish proof-of-principle for genomic prediction in insect biocontrol agents, using wing morphology traits in the parasitoid Nasonia vitripennis Walker (Pteromalidae) as a model. MethodsWe performed genomic prediction using a Genomic Best Linear Unbiased Prediction (GBLUP) model, using a total of 1,230 individuals with 8,639 SNPs generated by genotyping-by-sequencing (GBS). We used individuals from two generations from the outbred HVRx population, 717 individuals from generation G169 and 513 from generation G172. To assess genomic prediction accuracy, we used across- generation validation: forward validation for G172 from G169, backward in time validation for G169 from G172, and also 5-fold cross-validation, randomly using one fifth of the population as validiation and the others as training groups. ResultsFor size-related traits, including tibia length, wing length, width, and second moment wing area, the accuracy of genomic prediction was close to zero in both across-generation validations, but much higher in 5-fold cross-validation (ranging 0.54-0.68). For the shape-related trait wing aspect ratio, a high accuracy was found for all three validation strategies, with 0.47 for across-generation forward validation, 0.65 for across-generation backward validation, and 0.54 for 5-fold cross-validation. ConclusionPromising accuracies were observed for all traits in 5-fold cross-validation, but not in the across-generation validations. Overall, applying genomic selection in insect biocontrol agents with a relative small effective population size seems promising. However, factors such as the biology of insects, the techniques of phenotyping, and costs of large-scale genotyping still challenge the application of genomic selection to biocontrol agents.

genetics↗

Uncovering large effect loci and epistasis for lifespan in standing genetic variation in the fruit fly Drosophila melanogaster

Lifespan is a heritable trait with a polygenic architecture. Experimental evolution in combination with re-sequencing has often been used to identify candidate loci for lifespan in Drosophila melanogaster. Previous experiments showed that Drosophila populations experimentally evolved to increase late-life reproduction showed a correlated responses in development time, body size, but also lifespan. Subsequent whole genome sequencing allowed for the identification of candidate loci that correlated to lifespan differentiation. However, it remains difficult to assess whether candidate loci affect lifespan and to what extent such loci pleiotropically underpin multiple traits. Furthermore, recent studies indicate that lifespan effects of loci are often context dependent, but genotype-by-genotype interactions remain understudied. Therefore, here, we report on a study where we genotyped 3210 individuals for 32 candidate loci that emerged from our evolve and re-sequence experiment and tested, (1) whether these loci significantly affected lifespan, (2) the effect size of each locus, and, (3) how these loci mutually interact, i.e. determine the level of epistasis in moulding lifespan. Of the 32 loci, six showed significant main effect associations, of which three loci showed effects of 6.6 days difference in lifespan or larger, while the overall average lifespan was 41.7 days. Eight additional significant pairwise interactions between loci were found, of which four (single) main effects and one three-way interaction was significant. Lastly, we found that alleles that increased lifespan did not necessarily have higher frequencies in populations that showed increased lifespan, indicating that lifespan itself had not been the major target of selection. Our study indicates that individual genotyping following an evolve and re-sequencing study is essential to understand the mechanistic basis of polygenetic adaptation.

evolutionary biology↗

Insecticide resistance in Myzus persicae collected from sweet pepper

Populations of the green peach aphid, Myzus persicae, rapidly develop resistance to insecticides applied in agriculture, necessitating regular resistance monitoring of pest populations. Previous research identified two dominant multilocus genotypes (MLGs) in conventional (using insecticides + biological control agents) Dutch sweet pepper greenhouses: MLG-A under pymetrozine application and MLG-R under flonicamid application. This suggests positive selection for the genotypes by the application of these insecticides. However, no resistance of M. persicae to these insecticides has been reported yet. To investigate whether the insecticides were selectively driving the emergence of these specific MLGs, we compared the sensitivity of MLG-A and MLG-R to pymetrozine and flonicamid to that of six other genotypes from the same crop system. Additionally, we screened the M. persicae populations from Dutch sweet pepper greenhouses for known mutations conferring resistance to carbamates, pyrethroids, neonicotinoids, and tetronic and tetramic acid derivatives. Our results show that both MLG-A and MLG-R are less sensitive to pymetrozine compared to the other genotypes investigated. Furthermore, full mortality for MLG-R, the genotype least sensitive to flonicamid, was not achieved at the recommended field dose for this insecticide. Resistance mutations for carbamates and pyrethroids were prevalent among the MLGs, including MLG-A and MLG-R, while mutation A2226V, which is linked to resistance to tetronic and tetramic acid derivatives, was absent. Notably, the neonicotinoid resistance mutation R81T was found only in MLG-R, making this the northernmost detection of R81T in M. persicae to date. This study shows that various resistance mechanisms can accumulate in a single aphid genotype and that insecticides likely play a role in selecting for the dominant genotypes of M. persicae in conventional greenhouses.

genetics↗

Population genetic structure of the aphid pest Myzus persicae in organic and conventional greenhouses

Aphids display remarkable adaptability to pest control strategies and their partheno-genetic reproduction results in rapid numerical increase of higher-fitness clones. Con-sequently, a high prevalence of only a few clonal lines could indicate positive selection for these genotypes, potentially reflecting adaptation to pest control methods. Here, we investigated the clonal diversity and population genetic structure of the green peach aphid Myzus persicae using microsatellite markers, in both organic and conventionally managed Dutch sweet pepper greenhouses over four consecutive years. In total, 26 distinct multilocus genotypes (MLG) were detected, with higher clonal diversity in organic than in conventional greenhouses. Strikingly, a single MLG dominated conventional greenhouses in 2019, only to be completely replaced by a new dominant MLG by 2022. Whole-genome sequencing of 15 sampled lines -- seven sharing the same MLG and eighth with unique MLGs -- revealed that all aphids with the dominant MLG of 2019, collected from various locations and over multiple years, originated from a single parthenogenetic ancestor. Our findings indicate that the population genetic structure of M. persicae differs between organically and conventionally managed sweet pepper greenhouses. The presence of dominant MLGs in conventional crop systems may suggest positive selection or evolutionary forces such as a founder effect. Understanding the forces driving these differences in population genetic structure and their impact on the efficacy of biocontrol agents will further help us improve control strategies for M. persicae in greenhouse crops.

genetics↗

Improving the suppressive power of homing gene drive by co-targeting a distant-site female fertility gene

Gene drive technology has the potential to address major biological challenges, including the management of disease vectors, invasive species, and agricultural pests. After releasing individuals carrying the gene drive in the target population, suppression gene drives are designed to spread at a rapid rate and carry a recessive fitness cost, thus bringing about a decline in population size or even complete suppression. Well-studied homing suppression drives have been shown to be highly efficient in Anopheles mosquitoes and were successful in eliminating large cage populations. However, for other organisms, including Aedes mosquitoes, homing gene drives are so far too inefficient to achieve complete population suppression, mainly due to lower rates of drive conversion, which is the rate at which wild type alleles are converted into drive alleles. Low drive conversion is also a major issue in vertebrates, as indicated by experiments in mice. To tackle this issue, we propose a novel gene drive design that has two targets: a homing site where the drive is located and drive conversion takes place (with rescue for an essential gene), and a distant site for providing the fitness cost for population suppression (preferably a female fertility gene, for which no rescue is provided). We modeled this design and found that the two-target system allows suppression to occur over a much wider range of drive conversion efficiency. Specifically, in the new design, the suppressive power depends mostly on total gRNA cutting efficiency instead of just drive conversion efficiency, which is advantageous because cut rates are often substantially higher than drive conversion rates. We constructed a proof of concept in Drosophila melanogaster and show that both components of the gene drive function successfully. However, embryo drive activity from maternally deposited Cas9 as well as fitness costs for female drive heterozygotes both remain significant challenges for two-target and standard suppression drives. Overall, our improved gene drive design eases the development of strong homing suppression gene drives for many species where drive conversion is less efficient.

genetics↗

Genetic differentiation at extreme latitudes in the socially plastic sweat bee Halictus rubicundus

The sweat bee Halictus rubicundus is an important pollinator with a large latitudinal range and many potential barriers to gene flow. Alongside typical physical barriers, including mountain ranges and oceans, the climate may also impose restrictions on gene flow in this species. The climate influences voltinism and sociality in H. rubicundus, which is bivoltine and can nest socially at lower latitudes but can be univoltine and solitary in the north of its range and at higher altitudes where the climate is cooler. Variation in voltinism due to the climate may result in differences in phenology between populations across this species geographical range. Differences in phenology could limit gene flow, rendering populations at extreme latitudes genetically isolated and potentially more vulnerable to environmental stressors. A previous study found that the Irish Sea restricts gene flow in this species, but there was no evidence that differences in phenology had a similar effect as there was no genetic differentiation between H. rubicundus populations on mainland Britain. Here we extend the previous study to consider populations of H. rubicundus at extreme northern and southern latitudes in the UK. Using 12 microsatellite markers for genotyping, we found that bees from a population in the far north of Scotland were genetically differentiated from bees collected in Cornwall in the south-west of England. In contrast, bees collected across the Irish Sea in Northern Ireland showed genetic overlap with both the Scottish and Cornish bees. Our results suggest that when populations at extreme latitudes are considered, phenology and the climate may act alongside physical barriers such as the Scottish Highlands and the Irish Sea to restrict gene flow in H. rubicundus. We discuss the implications of our results for local adaptation in the face of rapidly changing selection pressures which are likely under climate change.

genetics↗

A single QTL with large effect is associated with female functional virginity in an asexual parasitoid wasp

During the transition from sexual to asexual reproduction, a suite of reproduction-related sexual traits become superfluous, and may be selected against if costly. Female functional virginity refers to asexual females resisting to mate or not fertilizing eggs after mating. These traits appear to be among the first that evolve during the gradual transition from sexual to asexual reproduction. The genetic basis of female functional virginity remains elusive. Previously, we reported that female functional virginity segregates as a single recessive locus in the asexual parasitoid wasp Asobara japonica. Here, we investigate the genetic basis of this trait by quantitative trait loci (QTL) mapping and candidate gene analyses. Consistent with the segregation of phenotypes, a single QTL of large effect was found spanning over 4.23 Mb and comprising at least 131 protein-coding genes, of which 15 featured sex-biased expression in the related sexual Asobara tabida. We speculate that two of these 15 genes may be of particular interest: CD151 antigen and nuclear pore complex protein Nup50. Overall, our results are consistent with a single gene or a cluster of linked genes underlying rapid evolution of female functional virginity in the transition to asexuality. Once a mutation for rejection to mate has swept through a population, the region comprising the gene(s) does not get smaller due to lack of recombination in asexuals.

evolutionary biology↗

Braconidae revisited: Bracon brevicornis genome showcases the potential of linked-read sequencing in identifying a putative complementary sex determiner gene

Bracon brevicornis is an ectoparasitoid of a wide range of larval-stage Lepidopterans, including several pests of important crops, such as the corn borer, Ostrinia nubilalis. It is also one of the earliest documented cases of complementary sex determination in Hymenoptera. Here, we present the linked-read genome of B. brevicornis, complete with an ab initio-derived annotation and protein comparisons with fellow braconids, Fopius arisanus and Diachasma alloem. We demonstrate the potential of linked-read assemblies in exploring regions of heterozygosity and search for structural and homology-derived evidence of the complementary sex determiner gene (csd).

genomics↗

Hybrid genome assembly and evidence-based annotation of the egg parasitoid and biological control agent Trichogramma brassicae

Trichogramma brassicae (Bezdenko) are egg parasitoids that are used throughout the world as biological control agents and in laboratories as model species. Despite this ubiquity, few genetic resources exist beyond COI, ITS2, and RAPD markers. Aided by a Wolbachia infection, a wild-caught strain from Germany was reared for low heterozygosity and sequenced in a hybrid de novo strategy, after which several assembling strategies were evaluated. The best assembly, derived from a DBG2OLC-based pipeline, yielded a genome of 235 Mbp made up of 1,572 contigs with an N50 of 556,663 bp. Following a rigorous ab initio-, homology-, and evidence-based annotation, 16,905 genes were annotated and functionally described. As an example of the utility of the genome, a simple ortholog cluster analysis was performed with sister species T. pretiosum, revealing over 6000 shared clusters and under 400 clusters unique to each species. The genome and transcriptome presented here provides an essential resource for comparative genomics of the commercially relevant genus Trichogramma, but also for research into molecular evolution, ecology, and breeding of T. brassicae.

genomics↗

Genomics of sex allocation in the parasitoid wasp Nasonia vitripennis

BackgroundWhilst adaptive facultative sex allocation has been widely studied at the phenotypic level across a broad range of organisms, we still know remarkably little about its genetic architecture. Here, we explore the genome-wide basis of sex ratio variation in the parasitoid wasp Nasonia vitripennis, perhaps the best studied organism in terms of sex allocation, and well known for its response to local mate competition (LMC). ResultsWe performed a genome-wide association study (GWAS) for single foundress sex ratios using iso-female lines derived from the recently developed outbred N. vitripennis laboratory strain HVRx. The iso-female lines capture a sample of the genetic variation in HVRx and we present them as the first iteration of the Nasonia vitripennis Genome Reference Panel (NVGRP 1.0). This panel provides an assessment of the standing genetic variation for sex ratio in the study population. Using the NVGRP, we discovered a cluster of 18 linked SNPs, encompassing 9 annotated loci associated with sex ratio variation. Furthermore, we found evidence that sex ratio has a shared genetic basis with clutch size on three different chromosomes. ConclusionsOur approach provides a thorough description of the quantitative genetic basis of sex ratio variation in Nasonia at the genome level and reveals a number of inter-related candidate loci underlying sex allocation regulation.

evolutionary biology↗

Jekyll or Hyde? The genome (and more) of Nesidiocoris tenuis, a zoophytophagous predatory bug that is both a biological control agent and a pest

Nesidiocoris tenuis (Reuter) is an efficient predatory biological control agent used throughout the Mediterranean Basin in tomato crops but regarded as a pest in northern European countries. Belonging to the family Miridae, it is an economically important insect yet very little is known in terms of genetic information - no published genome, population studies, or RNA transcripts. It is a relatively small and long-lived diploid insect, characteristics that complicate genome sequencing. Here, we circumvent these issues by using a linked-read sequencing strategy on a single female N. tenuis. From this, we assembled the 355 Mbp genome and delivered an ab initio, homology-based, and evidence-based annotation. Along the way, the bacterial "contamination" was removed from the assembly, which also revealed potential symbionts. Additionally, bacterial lateral gene transfer (LGT) candidates were detected in the N. tenuis genome. The complete gene set is composed of 24,688 genes; the associated proteins were compared to other hemipterans (Cimex lectularis, Halyomorpha halys, and Acyrthosiphon pisum), resulting in an initial assessment of unique and shared protein clusters. We visualised the genome using various cytogenetic techniques, such as karyotyping, CGH and GISH, indicating a karyotype of 2n=32 with a male-heterogametic XX/XY system. Additional analyses include the localization of 18S rDNA and unique satellite probes via FISH techniques. Finally, population genomics via pooled sequencing further showed the utility of this genome. This is one of the first mirid genomes to be released and the first of a mirid biological control agent, representing a step forward in integrating genome sequencing strategies with biological control research.

genomics↗

Genome-wide disruption of DNA methylation by 5-aza-2’-deoxycytidine in the parasitioid wasp Nasonia vitripennis

DNA methylation of cytosine residues across the genome influences how genes and phenotypes are regulated in a wide range of organisms. As such, understanding the role of DNA methylation and other epigenetic mechanisms has become very much a part of mapping genotype to phenotype, a major question in evolutionary biology. Ideally, we would like to manipulate DNA methylation patterns on a genome-wide scale, to help us to elucidate the role that epigenetic modifications play in phenotypic expression. Recently, the demethylating agent 5-aza-2-deoxycytidine (5-aza-dC; commonly used in the epigenetic treatment of certain cancers), has been deployed to explore the epigenetic regulation of a number of traits of interest to evolutionary ecologists, including facultative sex allocation in the parasitoid wasp Nasonia vitripennis. In a recent study, we showed that treatment with 5-aza-dC did not ablate the facultative sex allocation response in Nasonia, but shifted the patterns of sex allocation in a way predicted by genomic conflict theory. This was the first (albeit indirect) experimental evidence for genomic conflict over sex allocation facilitated by DNA methylation. However, that work lacked direct evidence of the effects of 5-aza-dC on DNA methylation, and indeed the effect of the chemical has since been questioned in Nasonia. Here, using whole-genome bisulphite sequencing of more than 4 million CpGs, across more than 11,000 genes, we demonstrate unequivocally that 5-aza-dC disrupts methylation on a large scale across the Nasonia vitripennis genome. We show that the disruption can lead to both hypo- and hyper-methylation, may vary across tissues and time of sampling, and that the effects of 5-aza-dC are context- and sequence specific. We conclude that 5-aza-dC does indeed have the potential to be repurposed as a tool for studying the role of DNA methylation in evolutionary ecology, whilst many details of its action remain to be discovered. Author SummaryShedding light on the mechanistic basis of phenotypes is a major aim in the field of evolutionary biology. If we understand how phenotypes are controlled at the molecular level, we can begin to understand how evolution has shaped that phenotype and conversely, how genetic architecture may constrain trait evolution. Epigenetic markers (such as DNA methylation) also influence phenotypic expression by regulating how and when genes are expressed. Recently, 5-aza-2-deoxycytidine (5-aza-dC), a hypomethylating agent used in the treatment of certain cancers, has been used to explore the epigenetic regulation of traits of interest to evolutionary ecologists. Previously, we used 5-aza-dC to validate a role for DNA methylation in facultative sex allocation behaviour in the parasitoid wasp Nasonia vitripennis. However, the direct effects of the chemical were not examined at that point and its efficacy in insects was questioned. Here, we demonstrate that 5-aza-dC disrupts DNA methylation on a genome-wide scale in a context- and sequence-specific manner and results in both hypo- and hyper-methylation. Our work demonstrates that 5-aza-dC has the potential to be repurposed as a tool for studying the role of DNA methylation in phenotypic expression.

evolutionary biology↗