Tritrophic effects mediate host suitability for two parasitoids of the carob moth (Lepidoptera: Pyralidae), infesting pistachio kernels.

The carob moth, Ectomyelois ceratoniae (Zeller) (Lepidoptera: Pyralidae), is an important pest of pistachio causing direct damage to nuts during ripening, and in storage. We examined the tritrophic effects of 5 pistachio cultivars, representing the full range of kernel suitability for E. ceratoniae, on 2 larval parasitoids, Habrobracon hebetor (Say) (Hymenoptera: Braconidae), a gregarious, synovigenic, idiobiont, ectoparasitoid, and Venturia canescens Gravenhorst (Hymenoptera: Ichneumonidae), a solitary, pro-ovigenic, koinobiont, endoparasitoid. The suitability of pistachio cultivars for development and reproduction of the 2 parasitoids was closely correlated, despite their divergent lifestyles and developmental patterns, and paralleled variation in kernel suitability for host larvae. Although kernels of the most resistant cultivar reduced demographic parameters of the parasitoids, the potential impact on numerical responses was minor. Successful biological control of carob moth within the vulnerable period of nut ripening will likely depend on the functional responses of earlier generations of adult parasitoids emerging from other fruit and nut crops, or those released in an augmentation program.

A chromosome scale assembly of the parasitoid wasp Venturia canescens provides insight into the process of virus domestication.

The parasitoid wasp Venturia canescens is an important biological control agent of stored products moth pests and serves as a model to study the function and evolution of domesticated endogenous viruses (DEVs). The DEVs discovered in V. canescens are known as virus-like particles (VcVLPs), which are produced using nudivirus-derived components and incorporate wasp-derived virulence proteins instead of packaged nucleic acids. Previous studies of virus-derived components in the V. canescens genome identified 53 nudivirus-like genes organized in six gene clusters and several viral pseudogenes, but how VcVLP genes are organized among wasp chromosomes following their integration in the ancestral wasp genome is largely unknown. Here, we present a chromosomal scale genome of V. canescens consisting of 11 chromosomes and 56 unplaced small scaffolds. The genome size is 290.8 Mbp with a N50 scaffold size of 24.99 Mbp. A high-quality gene set including 11,831 protein-coding genes were produced using RNA-Seq data as well as publicly available peptide sequences from related Hymenoptera. A manual annotation of genes of viral origin produced 61 intact and 19 pseudogenized nudivirus-derived genes. The genome assembly revealed that two previously identified clusters were joined into a single cluster and a total of 5 gene clusters comprising of 60 intact nudivirus-derived genes were located in three chromosomes. In contrast, pseudogenes are dispersed among 8 chromosomes with only 4 pseudogenes associated with nudivirus gene clusters. The architecture of genes encoding VcVLP components suggests it originates from a recent virus acquisition and there is a link between the processes of dispersal and pseudogenization. This high-quality genome assembly and annotation represents the first chromosome-scale assembly for parasitoid wasps associated with VLPs, and is publicly available in the National Center for Biotechnology Information Genome and RefSeq databases, providing a valuable resource for future studies of DEVs in parasitoid wasps.

The for gene as one of the drivers of foraging variations in a parasitic wasp.

Foraging behaviours encompass strategies to locate resources and to exploit them. In many taxa, these behaviours are driven by a major gene called for, but the mechanisms of gene regulation vary between species. In the parasitoid wasp Venturia canescens, sexual and asexual populations coexist in sympatry but differ in life-history traits, physiology and behaviours, which could impact their foraging strategies. Here, we explored the molecular bases underpinning divergence in behaviours by testing two mutually nonexclusive hypotheses: first, the divergence in the for gene correlates with differences in foraging strategies, and second, the latter rely on a divergence in whole-genome expression. Using comparative genomics, we showed that the for gene was conserved across insects considering both sequence and gene model complexity. Polymorphism analysis did not support the occurrence of two allelic variants diverging across the two populations, yet the asexual population exhibited less polymorphism than the sexual population. Sexual and asexual transcriptomes split sharply, with 10.9% differentially expressed genes, but these were not enriched in behaviour-related genes. We showed that the for gene was more highly expressed in asexual female heads than in sexual heads and that those differences correlate with divergence in foraging behaviours in our experiment given that asexuals explored the environment more and exploited more host patches. Overall, these results suggested that fine tuning of for gene expression between populations may have led to distinct foraging behaviours. We hypothesized that reproductive polymorphism and coexistence in sympatry of sexual and asexual populations specialized to different ecological niches via divergent optima on phenotypic traits could imply adaptation through different expression patterns of the for gene and at many other loci throughout the genome.

Conserved Viral Transcription Plays a Key Role in Virus-Like Particle Production of the Parasitoid Wasp Venturia canescens.

Nudiviruses are large double-stranded DNA viruses related to baculoviruses known to be endogenized in the genomes of certain parasitic wasp species. These wasp-virus associations allow the production of viral particles or virus-like particles that ensure wasp parasitism success within lepidopteran hosts. Venturia canescens is an ichneumonid wasp belonging to the Campopleginae subfamily that has endogenized nudivirus genes belonging to the Alphanudivirus genus to produce "virus-like particles" (Venturia canescens virus-like particles [VcVLPs]), which package proteic virulence factors. The main aim of this study was to determine whether alphanudivirus gene functions have been conserved following endogenization. The expression dynamics of alphanudivirus genes was monitored by a high throughput transcriptional approach, and the functional role of lef-4 and lef-8 genes predicted to encode viral RNA polymerase components was investigated by RNA interference. As described for baculovirus infections and for endogenized nudivirus genes in braconid wasp species producing bracoviruses, a transcriptional cascade involving early and late expressed alphanudivirus genes could be observed. The expression of lef-4 and lef-8 was also shown to be required for the expression of alphanudivirus late genes allowing correct particle formation. Together with previous literature, the results show that endogenization of nudiviruses in parasitoid wasps has repeatedly led to the conservation of the viral RNA polymerase function, allowing the production of viruses or viral-like particles that differ in composition but enable wasp parasitic success. IMPORTANCE This study shows that endogenization of a nudivirus genome in a Campopleginae parasitoid wasp has led to the conservation, as for endogenized nudiviruses in braconid parasitoid wasps, of the viral RNA polymerase function, required for the transcription of genes encoding viral particles involved in wasp parasitism success. We also showed for the first time that RNA interference (RNAi) can be successfully used to downregulate gene expression in this species, a model in behavioral ecology. This opens the opportunity to investigate the function of genes involved in other traits important for parasitism success, such as reproductive strategies and host choice. Fundamental data acquired on gene function in Venturia canescens are likely to be transferable to other parasitoid wasp species used in biological control programs. This study also renders possible the investigation of other nudivirus gene functions, for which little data are available.

Adaptations to different habitats in sexual and asexual populations of parasitoid wasps: a meta-analysis.

BACKGROUND: Coexistence of sexual and asexual populations remains a key question in evolutionary ecology. We address the question how an asexual and a sexual form of the parasitoid Venturia canescens can coexist in southern Europe. We test the hypothesis that both forms are adapted to different habitats within their area of distribution. Sexuals inhabit natural environments that are highly unpredictable, and where density of wasps and their hosts is low and patchily distributed. Asexuals instead are common in anthropic environments (e.g., grain stores) where host outbreaks offer periods when egg-load is the main constraint on reproductive output. METHODS: We present a meta-analysis of known adaptations to these habitats. Differences in behavior, physiology and life-history traits between sexual and asexual wasps were standardized in term of effect size (Cohen's d value; Cohen, 1988). RESULTS: Seeking consilience from the differences between multiple traits, we found that sexuals invest more in longevity at the expense of egg-load, are more mobile, and display higher plasticity in response to thermal variability than asexual counterparts. DISCUSSION: Thus, each form has consistent multiple adaptations to the ecological circumstances in the contrasting environments.

Diploid male production correlates with genetic diversity in the parasitoid wasp Venturia canescens: a genetic approach with new microsatellite markers.

Sex determination is ruled by haplodiploidy in Hymenoptera, with haploid males arising from unfertilized eggs and diploid females from fertilized eggs. However, diploid males with null fitness are produced under complementary sex determination (CSD), when individuals are homozygous for this locus. Diploid males are expected to be more frequent in genetically eroded populations (such as islands and captive populations), as genetic diversity at the csd locus should be low. However, only a few studies have focused on the relation between population size, genetic diversity, and the proportion of diploid males in the field. Here, we developed new microsatellite markers in order to assess and compare genetic diversity and diploid male proportion (DMP) in populations from three distinct habitat types - mainland, island, or captive -, in the parasitoid wasp Venturia canescens. Eroded genetic diversity and higher DMP were found in island and captive populations, and habitat type had large effect on genetic diversity. Therefore, DMP reflects the decreasing genetic diversity in small and isolated populations. Thus, Hymenopteran populations can be at high extinction risk due to habitat destruction or fragmentation.

Impact of fluctuating temperatures on development of the koinobiont endoparasitoid Venturia canescens.

The effect of temperature on the biology of Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) is well understood under constant temperature conditions, but less so under more natural, fluctuating conditions. Herein we studied the influence of fluctuating temperatures on biological parameters of V. canescens. Parasitized fifth-instar larvae of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) were reared individually in incubators at six fluctuating temperature regimes (15-19.5°C with a mean of 17.6°C, 17.5-22.5°C with a mean of 19.8°C, 20-30°C with a mean of 22.7°C, 22.5-27.5°C with a mean of 25°C, 25.5-32.5°C with a mean of 28.3°C and 28.5-33°C with a mean of 30°C) until emergence and death of V. canescens adults. Developmental time from parasitism to adult eclosion, adult longevity and survival were recorded at each fluctuating temperature regime. In principle, developmental time decreased with an increase of the mean temperature of the fluctuating temperature regime. Upper and lower threshold temperatures for total development were estimated at 34.9 and 6.7°C, respectively. Optimum temperature for development and thermal constant were 28.6°C and 526.3 degree days, respectively. Adult longevity was also affected by fluctuating temperature, as it was significantly reduced at the highest mean temperature (7.0 days at 30°C) compared to the lowest one (29.4 days at 17.6°C). Survival was low at all tested fluctuating temperatures, apart from mean fluctuating temperature of 25°C (37%). Understanding the thermal biology of V. canescens under more natural conditions is of critical importance in applied contexts. Thus, predictions of biological responses to fluctuating temperatures may be used in population forecasting models which potentially influence decision-making in IPM programs.

Host switching in a generalist parasitoid: contrasting transient and transgenerational costs associated with novel and original host species.

Parasitoids face challenges by switching between host species that influence survival and fitness, determine their role in structuring communities, influence species invasions, and affect their importance as biocontrol agents. In the generalist parasitoid, Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae), we investigated the costs in encapsulation, survival, and body size on juveniles when adult parasitoids switched from their original host, Plodia interpunctella (Hübner) (Lepidotera, Pyralidae) to a novel host, Ephestia kuehniella (Zeller) (Lepidoptera, Pyralidae), over multiple generations. Switching had an initial survival cost for juvenile parasitoids in the novel host, but increased survival occurred within two generations. Conversely, mortality in the original host increased. Body size, a proxy for fecundity, also increased with the number of generations in the novel host species, reflecting adaptation or maternal effects due to the larger size of the novel host, and therefore greater resources available to the developing parasitoid. Switching to a novel host appears to have initial costs for a parasitoid, even when the novel host may be better quality, but the costs rapidly diminish. We predict that the net cost of switching to a novel host for parasitoids will be complex and will depend on the initial reduction in fitness from parasitizing a novel host versus local adaptations against parasitoids in the original host.

Energy dynamics in a parasitoid foraging in the wild.

Although parasitoids are used widely as a biological models for understanding the evolution of animal behaviour, most studies have been constrained to the laboratory. The dearth of field studies has been compounded by the almost complete ignorance of the physiological parameters involved in foraging and dispersal, in particular of the energetic constraints imposed by resource limitation. We estimated the dynamics of carbohydrates and lipids reserves of Venturia canescens (Gravenhorst) females by releasing individuals of known nutritional status in a natural environment and recapturing them using host-containing traps. The recapture rate was around 30%. These results were compared with the reserves of caged animals kept under different experimental conditions (freshly emerged, starved to death, fed ad libitum and partially starved). Wild animals were also sampled in order to estimate the resource levels of the local population. The results show that: (i) wasps are able to maintain a nearly constant level of energy over an extended foraging period; (ii) V. canescens takes sugars in the field; and (iii) the lipid reserves accumulated during the larval life may be limiting as lipogenesis does not take place in adults even under conditions of high sugar availability. These results demonstrate that wasps can forage for hosts and food and disperse in this habitat for hours and days without running into a severe risk of energy limitation.

Differential cannibalism and population dynamics in a host-parasitoid system.

The effects of host cannibalism on a host-parasitoid system were explored through experiment and modelling. In individual encounters between parasitized and unparasitized Plodia interpunctella larvae, parasitized larvae were more likely to be cannibalized. Inclusion of this differential cannibalism into a simple Lotka-Volterra-type model of host-parasitoid population dynamics generates alternative stable states-including stable coexistence and extinction of the parasitoid - which depend on starting conditions. Possible mechanisms for differential cannibalism, and its implications for studies of host-parasitoid populations and biological control programmes are discussed.