Thelytokous Wasps Research Articles

Co-occurrence of thelytokous and bisexual Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) in a natural population

Trichogramma dendrolimi is one of the most successful biocontrol agents in China. However, an inundative condition is necessary to obtain acceptable parasitism effect. A good solution to this is the application of its thelytokous counterparts which unfortunately are scarce in field. We here report the first case of a natural T. dendrolimi population in China comprising both bisexual wasps and an extremely low proportion of thelytokous wasps. These two forms of T. dendrolimi are phylogenetically related based on the reconstructions of ITS-2 and COI genes. Also, the phylogenetic results suggested a potentially Wolbachia-drived ITS-2 variation. The expression of thelytoky was hardly affected by temperature, which might help control Asian corn borer and Dendrolimus punctatus. Wolbachia are responsible for current thelytoky according to phylogenetic analyses, antibiotic treatment and introgression experiment. We also present the third case of paternal sex ratio chromosome that restrains the expansion of Wolbachia. Moreover, the low frequency of thelytoky may be common in natural populations. Consequently if for biological control it is determined that a thelytokous strain is to be preferred, then large number of field collected females should be set up as isofemale lines, to detect the rare thelytoky.

Incomplete removal of Wolbachia with tetracycline has two-edged reproductive effects in the thelytokous wasp Encarsia formosa (Hymenoptera: Aphelinidae)

Wolbachia pipientis are intracellular endosymbionts that induce parthenogenesis in the parasitoid Encarsia formosa. Previous studies that focused on effects of Wolbachia on the wasp usually used tetracycline to remove Wolbachia without concern for the joint influences of tetracycline and Wolbachia. Here we treated the wasps (F0 lines) with tetracycline to produce offspring (F1 lines) which were not fed tetracycline to avoid antibiotic influence. The quantitative data and fluorescence in situ hybridization showed that Wolbachia titers were reduced but not totally removed. The Wolbachia that infected the male offspring were unpredictably detected. Low dose tetracycline enhanced the fertility of 2-day-old F0 wasps after 24 h of treatment; however, compared with controls, the oocyte load of 3- to 6-day-old tetracycline-treated wasps decreased day by day, and tetracycline reduced the longevity of the wasps. The fecundity of controls was significantly higher than that of the treated F1-10 and F1-20 generations. Gene expression of vitellogenin reflected the same trend as that of wasp fecundities in both F0 and F1 lines. Moreover, female offspring proportions of F0 and F1 lines were related to the titer of infected Wolbachia, demonstrating that Wolbachia titer affected the sex determination of E. formosa.

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 0.7–1.2 % male offspring. Seven to 39 % 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-015-0370-9) contains supplementary material, which is available to authorized users.

Omnia tempus habent: habitat-specific differences in olfactory learning and decision making in parasitic wasps

Olfactory learning is generally involved in the host-finding process in parasitic wasps. But the reliability of odour cues for predicting future host-finding success depends on the rate at which host-substrate associations are subject to variation within and between parasitoid generations. Since learning comes at physiological costs, we can expect animals to learn in a way that optimizes costs and benefits. The parasitic wasp Venturia canescens occurs in two reproductive modes that forage in different environments. We tested populations from both habitat types for learning rate, memory duration and speed of decision making and found considerable differences. Thelytokous wasps live in habitats with relatively stable host-substrate associations and might encounter hosts at a high rate. They showed a preference for a new odour after only a single experience. However, the response faded within 24h, even with spaced learning experiences. Arrhenotokous wasps live in habitats where hosts are scarce and are likely to be found on a variety of substrates. Like the thelytokous ones, arrhenotokous wasps learned a new odour after a single experience, but seemingly took long for information processing: one and four hours after an experience, a speed-accuracy trade-off became visible, while 24h after the experience, wasps decided quickly and in accordance with what they had learned. In addition, these wasps are likely to have developed an aversion response towards Geraniol in the CleanAir experiment. We conclude that the respective cognitive pattern can be attributed to the ecological circumstances of the wasp's natural habitat.

Characterization of intersex production in Trichogramma kaykai infected with parthenogenesis-inducing Wolbachia

Sexually aberrant individuals, displaying both male and female characteristics, are rare in occurrence but are documented throughout the animal kingdom. In parasitoid wasps of the genus Trichogramma, such individuals typically appear as a result of rearing Wolbachia-infected thelytokous wasps at high temperatures. Sexually aberrant Trichogramma have been referred to interchangeably in the literature as gynandromorphs, sexual mosaics and intersexes. However, accurately used, the terms "gynandromorph" and "sexual mosaic" describe an individual composed of a mixture of genetically distinct tissues corresponding to the sexual phenotypes observed, while "intersex" refers to an individual having a uniform genetic constitution but with some tissues exhibiting sexual phenotypes conflicting with the associated genotype. Here, we investigate the heat-induced production of sexually aberrant offspring by thelytokous Trichogramma kaykai. Aberrant individuals were rare, but each was characterized as one of 11 morphotypes ranging from very feminine to very masculine. Overall, the production of aberrant individuals increased with time from the onset of maternal oviposition. However, while the production of males also increased with time, the degree of masculinity of aberrant individuals did not; the different morphotypes appeared to be produced haphazardly. We conclude that the aberrant individuals produced by T. kaykai are actually intersexes and not gynandromorphs. The wasp's close association with Wolbachia and the absence of intersexes in uninfected populations allow us to discuss a possible origin of the condition.

Clonal genetic variation in a Wolbachia-infected asexual wasp: horizontal transmission or historical sex?

Wolbachia are endocellular bacteria known for manipulating the reproductive systems of many of their invertebrate hosts. Wolbachia are transmitted vertically from mother to offspring. In addition, new infections result from horizontal transmission between different host species. However, to what extent horizontal transmission plays a role in the spread of a new infection through the host population is unknown. Here, we investigate whether horizontal transmission of Wolbachia can explain clonal genetic variation in natural populations of Leptopilina clavipes, a parasitoid wasp infected with a parthenogenesis-inducing Wolbachia. We assessed variance of markers on the nuclear, mitochondrial and Wolbachia genomes. The nuclear and mitochondrial markers displayed significant and congruent variation among thelytokous wasp lineages, showing that multiple lineages have become infected with Wolbachia. The alternative hypothesis in which a single female became infected, the daughters of which mated with males (thus introducing nuclear genetic variance) cannot account for the presence of concordant variance in mtDNA. All Wolbachia markers, including the hypervariable wsp gene, were invariant, suggesting that only a single strain of Wolbachia is involved. These results show that Wolbachia has transferred horizontally to infect multiple female lineages during the early spread through L.clavipes. Remarkably, multiple thelytokous lineages have persisted side by side in the field for tens of thousands of generations.

A Comparative Study on the Functional Response ofWolbachia-Infected and Uninfected Forms of the Parasitoid WaspTrichogramma brassicae

Trichogramma species (Hymenoptera: Trichogrammatidae) are haplo-diploid egg parasitoids that are frequently used as biological control agents against lepidopteran pests. These wasps display two reproductive modes, including arrhenotoky (bisexuality) and thelytoky (unisexuality). Thelytokous forms are often associated with the presence of endosymbiotic Wolbachia bacteria. The use of thelytokous wasps has long been considered as a way to enhance the efficacy of biological control. The present study investigates the potential of a thelytokous Wolbachia-infected and an arrhenotokous uninfected Trichogramma brassicae Bezdenko strain as inundative biocontrol agents by evaluating their functional response towards different egg densities of the factitious host, the Angoumois grain moth, Sitotroga cerealella (Olivier) (Lepidoptera: Gelechiidae). The results revealed a type II functional response for both strains in which parasitism efficiency decreases with host egg density because of an increasing host handling time. A model with an indicator variable was used to compare the parameters of Holling's disc equation in different data sets. It was demonstrated that the two strains did not differ in host attack rate. However, the Wolbachia-infected strain did have an increased host handling time when compared to the bisexual strain. Some applied aspects of the findings are discussed.

Odor‐mediated patch choice in the parasitoid Venturia canescens: temporal decision dynamics

AbstractParasitoids foraging for hosts in a heterogeneous environment would greatly benefit if they could decide already from a distance in which areas search for resources would be most profitable and to avoid areas of low fitness returns. Interestingly, the temporal dynamics of the decision process in parasitoid patch choice have rarely been investigated. In a Y‐tube olfactometer, we tested whether thelytokous and arrhenotokous females of the parasitoid wasp Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) respond to differences in cues indicating the quality of a host‐containing patch and choose more profitable patches. Special attention was given to the time it took females to make their choices (patch choice time) when differences in patch quality were either qualitative (absence vs. presence of hosts and kairomone) or quantitative (various concentrations of hosts and kairomone, and presence of competitors). We found that both thelytokous and arrhenotokous wasps only chose the higher‐quality patch based on odor cues when the difference was qualitative. When patches differed only with respect to the number of hosts, or the presence or absence of competing female parasitoids, no significant preference could be found in females of either strain of the parasitoid. In contrast, both the time until females reached the junction of the Y‐tube olfactometer (response time) and the time until females decided for either patch (decision time) varied with parasitoid strain and odor treatment. Thelytokous wasps were faster than arrhenotokous wasps in their response time and in their decision time. However, females of both strains responded faster with increasing number of total hosts releasing kairomone. Yet, decision time for patches did not significantly vary as a function of patch quality offered to Venturia wasps.

Differential energy allocation as an adaptation to different habitats in the parasitic wasp Venturia canescens

Environmental pressures are expected to favour organisms that optimally allocate metabolic resources to reproduction and survival. We studied the resource allocation strategies and the associated tradeoffs in the parasitoid wasp Venturia canescens, and their adaptation to the characteristics of the environment. In this species, individuals of two reproductive modes coexist in the same geographical locations, but they mainly occur in distinct habitats. Thelytokous (asexual) wasps are mostly found in anthropogenic habitats, where hosts tend to aggregate and food is absent. Arrhenotokous (sexual) wasps are exclusively found in natural habitats, where hosts are scattered and food is present. We analysed (1) the quantity of energy stored during ontogeny, (2) the tradeoff between reproduction and survival, by measuring egg load and longevity and (3) the host patch exploitation behaviour of the wasps at emergence. Arrhenotokous wasps emerged with more metabolic resources than thelytokous ones, especially glycogen, a nutrient that could be used for flying in search of hosts and/or food. Thelytokous wasps allocated more energy than arrhenotokous wasps to egg production: this would allow them to parasitize more hosts. The tradeoff between egg production and longevity was not revealed within reproductive modes, but when comparing them. At emergence, arrhenotokous wasps tended to exploit host patches less thoroughly than thelytokous wasps, suggesting that by leaving the host patch, they search for food. The results clearly showed adaptations to the characteristics of habitats preferentially inhabited by the two reproductive modes, and suggested a mechanism that facilitates their coexistence in natural conditions.

Manipulation of oviposition choice of the parasitoid wasp,Encarsia pergandiella, by the endosymbiotic bacteriumCardinium

Reproductive manipulations of hosts by maternally inherited bacterial endosymbionts often result in an increase in the proportion of infected female hosts in the population. When this involves the conversion of incipient males to genetic or functional females, it presents unique difficulties for symbionts invading hosts with sex-specific reproductive behaviours, such as the autoparasitic Encarsia pergandiella. In sexual forms of this species, female eggs are laid in whitefly nymphs and male eggs are laid in conspecific or heterospecific parasitoids developing within the whitefly cuticle. Further, eggs laid in the 'wrong' host do not ordinarily complete development. This study explored the role of a bacterial symbiont, Cardinium, in manipulating oviposition behaviour in a thelytokous population of E. pergandiella. Oviposition choice was measured by the number and location of eggs deposited by both infected and uninfected adult waSPS in arenas containing equal numbers of hosts suitable for the development of male and female waSPS. Uninfected waSPS included antibiotic-treated female waSPS and (untreated) daughters of antibiotic-treated female waSPS. The choices of waSPS in the thelytokous population treatments were compared with those of a conspecific sexual population. We found that offspring of antibiotic-cured thelytokous waSPS reverted to the behaviour of unmated sexual waSPS, laying their few eggs almost exclusively in hosts appropriate for male eggs. Infected thelytokous waSPS distributed their eggs approximately evenly between host types, much like mated sexual female waSPS. The antibiotic-treated female waSPS exhibited choices intermediate to waSPS in the other two treatments. The change in the observed behaviour appears sufficient to allow invasion and persistence of Cardinium in sexual populations. Lastly, our results suggest a reduction in host discrimination as a possible mechanism by which Cardinium influences this change.

Different habitats, different habits? Response to foraging information in the parasitic wasp Venturia canescens

The parasitic wasp, Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae), has two reproductive modes, namely, thelytoky or arrhenotoky, and occurs in habitats with highly variable or relatively stable host abundances, respectively. Since information processing is costly, we expected that information indicating resource availability would be mainly used by the thelytokous wasps and less so by the arrhenotokous type. This idea was explored by two different approaches. In a study on patch-time allocation, we used females from ten populations and measured patch-residence times of individuals that visited multiple patches at different encounter rates. In a more detailed approach, thelytokous and arrhenotokous females from a single location were observed continuously while foraging, and all behaviors were recorded. Wasps of both reproductive modes (i.e., both habitat types) used information for the assessment of habitat quality. However, the way that the information was used differed between them. Whereas thelytokous females used foraging information to maximize their efficiency at high patch-encounter rates, arrhenotokous females merely reduced the number of offspring produced without changing patch times. The behavior of the arrhenotokous females should result in a spreading of offspring across the habitat and, thus, reduced sib-mating. The foraging strategy of these wasps might therefore be an adaptation to reduce costs associated with inbreeding.

Gene flow between arrhenotokous and thelytokous populations of Venturia canescens (Hymenoptera).

In the solitary parasitoid wasp Venturia canescens both arrhenotokously (sexual) and thelytokously (parthenogenetical) reproducing individuals occur sympatrically. We found in the laboratory that thelytokous wasps are able to mate, receive and use sperm of arrhenotokous males. Using nuclear (amplified fragment length polymorphism, virus-like protein) and mitochondrial (restriction fragment length polymorphism) markers, we show the occurrence of gene flow from the arrhenotokous to the thelytokous mode in the field. Our results reinforce the paradox of sex in this species.

Geographical distribution and genetic relatedness of sympatrical thelytokous and arrhenotokous populations of the parasitoid Venturia canescens (Hymenoptera)

Abstract Theory predicts that asexual reproduction has a competitive advantage over sexual reproduction because of the twofold cost of producing males. Few systems are suitable for directly testing this prediction. In the solitary parasitoid wasp Venturia canescens both arrhenotokously (sexual) and thelytokously (asexual) reproducing individuals occur sympatrically. We sampled 922 wasps from 22 localities along the coast of south-eastern France. Thelytokous wasps were less abundant (23%) than arrhenotokous wasps and were almost always found in sympatry with arrhenotokous ones. An analysis of genetic relatedness using amplified fragment length polymorphism (AFLP) markers showed the existence of a widespread thelytokous clone. In addition, a few thelytokous individuals were found to be closely related to arrhenotokous ones and vice versa. These data suggest the occurrence of occasional gene flow between both reproductive modes and/or recurrent origin of thelytokous clones from coexisting arrhenotokous populations in the area. The results are discussed in the context of the paradox of sex.

Host Deprivation Effect on Reproduction and Survival of Wolbachia-Infected and Uninfected Trichogramma kaykai Pinto & Stouthamer (Hymenoptera: Trichogrammatidae)

The effect of temporary host deprivation on the reproduction and survival of Trichogramma kaykai Pinto & Stouthamer was determined by comparing a Wolbachia-infected (thelytokous) and an arrhenotokous line of T. kaykai when reared on Trichoplusia ni Hubner (Lepidoptera: Noctuidae) eggs. The progeny produced by both arrhenotokous and thelytokous females decreased with increasing periods of host deprivation, but arrhenotokous females produced significantly more progeny than thelytokous females regardless of the period of host deprivation (0 to 10 days). They also produced more daughters. Host deprivation did not affect brood size within arrhenotokous or thelytokous lines. However, fewer wasps emerged from hosts parasitized by Wolbachia-infected than by uninfected T. kaykai. A direct relationship existed between longevity and the length of time an arrhenotokous or a thelytokous wasp was deprived of hosts. The longer the deprivation period, the longer they lived. However, arrhenotokous females lived longer than their thelytokous counterparts. These findings showed that temporary host deprivation (up to 10 days) affected both reproductive forms similarly. However, in all conditions studied arrhenotokous wasps produced more progeny and lived longer than their thelytokous counterparts.

Biological Control Potential of Wolbachia -infected Versus Uninfected Wasps: Laboratory and Greenhouse Evaluation of Trichogramma cordubensis and T. deion Strains

The effects of thelytoky-inducing Wolbachia ( f -proteobacteria) on Trichogramma cordubensis and T. deion (Hymenoptera, Trichogrammatidae) were studied in laboratory and greenhouse conditions. One infected (thelytokous, all female) line of each wasp species was compared with its conspecific uninfected (arrhenotokous, sexual) counterpart for several fecundity and dispersal traits. Arrhenotokous lines had a higher fecundity than their thelytokous counterparts, which suggests that Wolbachia negatively affect the fecundity of Trichogramma females. The arrhenotokous females dispersed more in the laboratory than their thelytokous counterparts. In the greenhouse, the opposite effect or no difference between lines was found, indicating that the laboratory set-up used to measure dispersal is not useful to predict relative dispersal of the females in the greenhouse. Calculations show that by releasing 100 adult wasps of both lines, thus including arrhenotokous males in the sexual line, more eggs are parasitized by the thelytokous wasps. Therefore, in spite of their lower individual female fecundity, thelytokous lines have a better potential for biological control than their arrhenotokous counterparts.