Bart A. Pannebakker

Development of a Nasonia vitripennis outbred laboratory population for genetic analysis

The parasitoid wasp genus Nasonia has rapidly become a genetic model system for developmental and evolutionary biology. The release of its genome sequence led to the development of high‐resolution genomic tools, for both interspecific and intraspecific research, which has resulted in great advances in understanding Nasonia biology. To further advance the utility of Nasonia vitripennis as a genetic model system and to be able to fully exploit the advantages of its fully sequenced and annotated genome, we developed a genetically variable and well‐characterized experimental population. In this study, we describe the establishment of the genetically diverse HVRx laboratory population from strains collected from the field in the Netherlands. We established a maintenance method that retains genetic variation over generations of culturing in the laboratory. As a characterization of its genetic composition, we provide data on the standing genetic variation and estimate the effective population size (Ne) by microsatellite analysis. A genome‐wide description of polymorphism is provided through pooled resequencing, which yielded 417 331 high‐quality SNPs spanning all five Nasonia chromosomes. The HVRx population and its characterization are freely available as a community resource for investigators seeking to elucidate the genetic basis of complex trait variation using the Nasonia model system.

The Transcriptomic Basis of Oviposition Behaviour in the Parasitoid Wasp Nasonia vitripennis

Linking behavioural phenotypes to their underlying genotypes is crucial for uncovering the mechanisms that underpin behaviour and for understanding the origins and maintenance of genetic variation in behaviour. Recently, interest has begun to focus on the transcriptome as a route for identifying genes and gene pathways associated with behaviour. For many behavioural traits studied at the phenotypic level, we have little or no idea of where to start searching for “candidate” genes: the transcriptome provides such a starting point. Here we consider transcriptomic changes associated with oviposition in the parasitoid wasp Nasonia vitripennis. Oviposition is a key behaviour for parasitoids, as females are faced with a variety of decisions that will impact offspring fitness. These include choosing between hosts of differing quality, as well as making decisions regarding clutch size and offspring sex ratio. We compared the whole-body transcriptomes of resting or ovipositing female Nasonia using a “DeepSAGE” gene expression approach on the Illumina sequencing platform. We identified 332 tags that were significantly differentially expressed between the two treatments, with 77% of the changes associated with greater expression in resting females. Oviposition therefore appears to focus gene expression away from a number of physiological processes, with gene ontologies suggesting that aspects of metabolism may be down-regulated during egg-laying. Nine of the most abundant differentially expressed tags showed greater expression in ovipositing females though, including the genes purity-of-essence (associated with behavioural phenotypes in Drosophila) and glucose dehydrogenase (GLD). The GLD protein has been implicated in sperm storage and release in Drosophila and so provides a possible candidate for the control of sex allocation by female Nasonia during oviposition. Oviposition in Nasonia therefore clearly modifies the transcriptome, providing a starting point for the genetic dissection of oviposition.

It is time to bridge the gap between exploring and exploiting: prospects for utilizing intraspecific genetic variation to optimize arthropods for augmentative pest control – a review

Intraspecific genetic variation in arthropods is often studied in the context of evolution and ecology. Such knowledge, however, can also be very usefully applied to biological pest control. Selection of genotypes with optimal trait values may be a powerful tool to develop more effective biocontrol agents. Although it has repeatedly been proposed, this approach is still hardly applied in the current commercial development of arthropod agents for pest control. In this perspective study, we call to take advantage of the increasing knowledge on the genetics underlying intraspecific variation to improve biological control agents. We argue that it is timely now because at present both the need and the technical possibilities for implementation exist, as there is (1) increased economic importance of biocontrol, (2) reduced availability of exotic biocontrol agents due to stricter legislation, and (3) increased availability of genetic information on non‐model species. We present a step‐by‐step approach towards the exploitation of intraspecific genetic variation for biocontrol, outline that knowledge of the underlying genetic mechanisms is essential for success, and indicate how new molecular techniques can facilitate this. Finally, we exemplify this procedure by two case studies, one focussing on a target trait – offspring sex ratio – across species of hymenopteran parasitoids, and the other on a target species – the two‐spot ladybird beetle – where wing length and body colouration can be optimized for aphid control. With this overview, we aim to inspire scientific researchers and biocontrol agent producers to start collaborating on the use of genetic variation for the improvement of natural enemies.

Absence of complementary sex determination in the parasitoid wasp genus Asobara (Hymenoptera: Braconidae)

An attractive way to improve our understanding of sex determination evolution is to study the underlying mechanisms in closely related species and in a phylogenetic perspective. Hymenopterans are well suited owing to the diverse sex determination mechanisms, including different types of Complementary Sex Determination (CSD) and maternal control sex determination. We investigated different types of CSD in four species within the braconid wasp genus Asobara that exhibit diverse life-history traits. Nine to thirteen generations of inbreeding were monitored for diploid male production, brood size, offspring sex ratio, and pupal mortality as indicators for CSD. In addition, simulation models were developed to compare these observations to predicted patterns for multilocus CSD with up to ten loci. The inbreeding regime did not result in diploid male production, decreased brood sizes, substantially increased offspring sex ratios nor in increased pupal mortality. The simulations further allowed us to reject CSD with up to ten loci, which is a strong refutation of the multilocus CSD model. We discuss how the absence of CSD can be reconciled with the variation in life-history traits among Asobara species, and the ramifications for the phylogenetic distribution of sex determination mechanisms in the Hymenoptera.

Population-level consequences of complementary sex determination in a solitary parasitoid

BackgroundSex determination mechanisms are known to be evolutionarily labile but the factors driving transitions in sex determination mechanisms are poorly understood. All insects of the Hymenoptera are haplodiploid, with males normally developing from unfertilized haploid eggs. Under complementary sex determination (CSD), diploid males can be produced from fertilized eggs that are homozygous at the sex locus. Diploid males have near-zero fitness and thus represent a genetic load, which is especially severe under inbreeding. Here, we study mating structure and sex determination in the parasitoid Cotesia vestalis to investigate what may have driven the evolution of two complementary sex determination loci in this species.ResultsWe genotyped Cotesia vestalis females collected from eight fields in four townships in Western Taiwan. 98 SNP markers were developed by aligning Illumina sequence reads of pooled DNA of eight different females against a de novo assembled genome of C. vestalis. This proved to be an efficient method for this non-model species and provides a resource for future use in related species. We found significant genetic differentiation within the sampled population but variation could not be attributed to sampling locations by AMOVA. Non-random mating was detected, with 8.1% of matings between siblings. Diploid males, detected by flow cytometry, were produced at a rate of 1.4% among diploids.ConclusionsWe think that the low rate of diploid male production is best explained by a CSD system with two independent sex loci, supporting laboratory findings on the same species. Fitness costs of diploid males in C. vestalis are high because diploid males can mate with females and produce infertile triploid offspring. This severe fitness cost of diploid males combined with non-random mating may have resulted in evolution from single locus CSD to CSD with two independent loci.

Diploid males support a two-step mechanism of endosymbiont-induced thelytoky in a parasitoid wasp

BackgroundHaplodiploidy, where females develop from diploid, fertilized eggs and males from haploid, unfertilized eggs, is abundant in some insect lineages. Some species in these lineages reproduce by thelytoky that is caused by infection with endosymbionts: infected females lay haploid eggs that undergo diploidization and develop into females, while males are very rare or absent. It is generally assumed that in thelytokous wasps, endosymbionts merely diploidize the unfertilized eggs, which would then trigger female development.ResultsWe found that females in the parasitoid wasp Asobara japonica infected with thelytoky-inducing Wolbachia produce 0.7–1.2xa0% male offspring. Seven to 39xa0% of these males are diploid, indicating that diploidization and female development can be uncoupled in A. japonica. Wolbachia titer in adults was correlated with their ploidy and sex: diploids carried much higher Wolbachia titers than haploids, and diploid females carried more Wolbachia than diploid males. Data from introgression lines indicated that the development of diploid individuals into males instead of females is not caused by malfunction-mutations in the host genome but that diploid males are most likely produced when the endosymbiont fails to activate the female sex determination pathway. Our data therefore support a two-step mechanism by which endosymbionts induce thelytoky in A. japonica: diploidization of the unfertilized egg is followed by feminization, whereby each step correlates with a threshold of endosymbiont titer during wasp development.ConclusionsOur new model of endosymbiont-induced thelytoky overthrows the view that certain sex determination mechanisms constrain the evolution of endosymbiont-induced thelytoky in hymenopteran insects. Endosymbionts can cause parthenogenesis through feminization, even in groups in which endosymbiont-diploidized eggs would develop into males following the hosts’ sex determination mechanism. In addition, our model broadens our understanding of the mechanisms by which endosymbionts induce thelytoky to enhance their transmission to the next generation. Importantly, it also provides a novel window to study the yet-poorly known haplodiploid sex determination mechanisms in haplodiploid insects.

Genetics of decayed sexual traits in a parasitoid wasp with endosymbiont-induced asexuality.

Trait decay may occur when selective pressures shift, owing to changes in environment or life style, rendering formerly adaptive traits non-functional or even maladaptive. It remains largely unknown if such decay would stem from multiple mutations with small effects or rather involve few loci with major phenotypic effects. Here, we investigate the decay of female sexual traits, and the genetic causes thereof, in a transition from haplodiploid sexual reproduction to endosymbiont-induced asexual reproduction in the parasitoid wasp Asobara japonica. We take advantage of the fact that asexual females cured of their endosymbionts produce sons instead of daughters, and that these sons can be crossed with sexual females. By combining behavioral experiments with crosses designed to introgress alleles from the asexual into the sexual genome, we found that sexual attractiveness, mating, egg fertilization and plastic adjustment of offspring sex ratio (in response to variation in local mate competition) are decayed in asexual A. japonica females. Furthermore, introgression experiments revealed that the propensity for cured asexual females to produce only sons (because of decayed sexual attractiveness, mating behavior and/or egg fertilization) is likely caused by recessive genetic effects at a single locus. Recessive effects were also found to cause decay of plastic sex-ratio adjustment under variable levels of local mate competition. Our results suggest that few recessive mutations drive decay of female sexual traits, at least in asexual species deriving from haplodiploid sexual ancestors.

Drawbacks and benefits of hygienic behavior in honey bees (Apis mellifera L.): a review

The hygienic behavior of honey bee workers contributes to the social immunity of colonies. The ability of workers to detect and remove unhealthy or dead brood prevents the transmission of brood diseases inside the colony. Over the last five decades, this trait has been extensively studied and improved in several research and breeding programs. Given the strong interest for hygienic behavior, we here review the costs and benefits associated with this trait, extending preceding reviews on this subject from the late 1990s. Since the 1990s, there have been no major new insights on the efficiency of this behavior against American foulbrood and chalkbrood. However, the number of publications on hygienic behavior against the mite Varroa destructor has considerably increased, fueling the debate regarding the efficiency of hygienic behavior against this parasite. Breeding programs have shown that selection for a specific trait might also impact other traits. Thus, we also review the cost of trade-offs between hygienic behavior and other economically important traits for bee breeders. Overall, the benefits of hygienic behavior seem to largely outweigh its costs for both colonies and bee breeders.

The chemical basis of mate recognition in two parasitoid wasp species of the genus Nasonia

To recognize ones mate is essential for all sexually reproducing animals. In insects, mate recognition is often based on chemical cues such as hydrocarbons which are distributed over the insects cuticle. In the parasitoid wasp genus Nasonia (Hymenoptera: Pteromalidae), interspecific mating possibly occurs in microsympatry between Nasonia vitripennis Walker and Nasonia giraulti Darling despite post‐zygotic isolation mechanisms preventing hybridization. Males of N. vitripennis are known to equally court con‐ and heterospecific females, which they recognize by means of cuticular hydrocarbons. A recent study surprisingly showed that this might not be the case in N. giraulti, leaving open how males of this species achieve the recognition of mating partners. In this study, we investigated chemical mate recognition in N. giraulti in more detail and compared observed behaviors with behaviors of N. vitripennis by conducting experiments with both species concurrently and under the same experimental conditions. We disentangled the role of female‐derived non‐polar cuticular lipids – i.e., cuticular hydrocarbons – and more polar cuticular lipids in the ability of males to recognize con‐ and heterospecific females. In addition, we tested whether females of the two species discriminate similarly between con‐ and heterospecific males. We demonstrate that, in contrast to N. vitripennis, males of N. giraulti prefer live conspecific females over heterospecific ones. Furthermore, in contrast to N. vitripennis, mate recognition in N. giraulti males is not based on cuticular hydrocarbons, but rather involves other chemical messengers, presumably more polar cuticular lipids. In both species, discrimination against heterospecific males decreases with female age.

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 many genes and phenotypes are regulated. 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 elucidate the role of epigenetic modifications 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. Recently, we showed that treatment with 5-aza-dC shifted patterns of sex allocation as predicted by genomic conflict theory in the parasitoid wasp Nasonia vitripennis. This was the first (albeit indirect) experimental evidence for genomic conflict over sex allocation facilitated by DNA methylation. However, this work lacked confirmation of the effects of 5-aza-dC on DNA methylation, drawing commentary on the efficacy of 5-aza-dC in a novel system. Here, using whole-genome bisulphite sequencing, we demonstrate unequivocally that 5-aza-dC disrupts methylation across the Nasonia vitripennis genome. We show that disruption leads 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 has the potential to be repurposed as a tool in evolutionary ecology for studying the role of DNA methylation.