A maternal gene mutation correlates with an ovary phenotype in a parthenogenetic wasp population.

Endoparasitoid wasps rely on maternal protein secretions, including viruses and virus-like particles (VLPs), to overcome host defense reactions. In the ichneumonid Venturia canescens, VLPs are assembled in the nuclei of ovarian calyx gland cells, secreted into the lumen of the gland, and eventually transmitted into the host caterpillar together with the parasitoid egg. One of the genes coding for VLP proteins, termed VLP1, exists in two alleles producing two structurally different proteins. Here we describe the establishment and initial phenotypic characterisation of two parthenogenetic laboratory strains, which differ in VLP1 as well as in other genetic markers. A comparison of calyx tissues from the two strains revealed morphological differences that seem to affect egg movement from the ovarioles into the oviduct. The observed histological changes are correlated with differences in egg maturation and embryonic development causing a delay in larval hatching in one of the strains. Under conditions that favour superparasitism, the two strains differ in the number of offspring produced.     

Genetic analysis of two distinct reproductive strategies in sexual and asexual field populations of an endoparasitic wasp,Venturia canescens

Asexual (thelytokous) females of the parasitoid Venturia canescens, which develop inside another insect, exhibit evolutionarily stable mixtures of life-history strategies, allowing two genetically distinct wasp lines to coexist sympatrically on the same host resources. Since the two asexual lines differ in a virus-like particle protein-coding gene (VLP1), the question is whether the VLP1 gene is genetically associated with the phenotype. The recent isolation of facultative sexual (arrhenotokous) and asexual V. canescens strains from the same location in Southern France has enabled an investigation of the genetic basis for the observed phenotypic differences, by comparing the two asexual lines with the corresponding homozygous VLP1 genotypes in arrhenotokous strains. This analysis showed similar patterns of morphological and functional differences exist in the ovaries of the two asexual VLP1 lines and in the two homozygous VLP1 genotypes from the field, suggesting that the VLP1 gene alteration either causes the ovarian phenotype or is genetically closely linked to the putative gene. However, the VLP1-gene may not be the only gene contributing to the phenotypic effects observed in the asexual lines. Although the two VLP1-alleles segregate with the relative differences in the ovary distribution of eggs, the absolute egg numbers differ in the corresponding asexual and sexual genotypes. This suggests that an additional unlinked gene may be involved in the transfer of eggs from the ovarioles into the oviduct.     

Lifetime gains of host-feeding in a synovigenic parasitic wasp

Abstract. Understanding behavioural decisions relative to host use for feeding or reproduction by foraging parasitoids requires not only the study of metabolic pathways followed by nutrients, but also the quantification of lifetime fitness gains of each alternative behaviour. By using a combination of observational and manipulative approaches, the lifetime host‐feeding gains are measured both in terms of fecundity and longevity in the parasitoid Eupelmus vuilletti. Host‐feeding increases both egg production and longevity. The increase in fecundity is mainly determined by the amount of lipids obtained whereas the lifespan extension is mainly determined by carbohydrates. Proteins obtained through host‐feeding have been implicated previously in egg production by parasitoids but protein intake has no effect on fecundity and longevity in E. vuilletti. The amount of nutrients gained through host‐feeding and invested in eggs is variable and changes over the lifetime of the animal. Therefore, lifetime feeding gains are best understood through the construction of dynamical budgets running over the entire lifespan of an insect.     

Mean reproductive traits of fungal assemblages are correlated with resource availability

Organisms have evolved a fascinating variety of strategies and organs for successful reproduction. Fruit bodies are the reproductive organ of fungi and vary considerably in size and shape among species. Our understanding of the mechanisms underlying the differences in fruit body size among species is still limited. Fruit bodies of saprotrophic fungi are smaller than those of mutualistic ectomycorrhizal fungi. If differences in fruit body size are determined by carbon acquisition, then mean reproductive traits of saprotrophic and ectomycorrhizal fungi assemblages should vary differently along gradients of resource availability as carbon acquisition seems more unpredictable and costly for saprotrophs than for ectomycorrhizal fungi. Here, we used 48 local inventories of fungal fruit bodies (plot size: 0.02 ha each) sampled along a gradient of resource availability (growing stock) across 3 years in the Bavarian Forest National Park in Germany to investigate regional and local factors that might influence the distribution of species with different reproductive traits, particularly fruit body size. As predicted, mean fruit body size of local assemblages of saprotrophic fungi was smaller than expected from the distribution of traits of the regional species pool across central and northern Europe, whereas that of ectomycorrhizal fungi did not differ from random expectation. Furthermore and also as expected, mean fruit body size of assemblages of saprotrophic fungi was significantly smaller than for assemblages of ectomycorrhizal species. However, mean fruit body sizes of not only saprotrophic species but also ectomycorrhizal species increased with resource availability, and the mean number of fruit bodies of both assemblages decreased. Our results indicate that the differences in carbon acquisition between saprotrophs and ectomycorrhizal species lead to differences in basic reproductive strategies, with implications for the breadth of their distribution. However, the differences in resource acquisition cannot explain detailed species distribution patterns at a finer, local scale based on their reproductive traits.     

Niche differentiation and colonization of a novel environment by an asexual parasitic wasp

How do asexual taxa become adapted to a diversity of environments, and how do they persist despite changing environmental conditions? These questions are linked by their mutual focus on the relationship between genetic variation, which is often limited in asexuals, and the ability to respond to environmental variation. Asexual taxa originating from a single ancestor present a unique opportunity to assess rates of phenotypic and genetic change when access to new genetic variation is limited to mutation. Diachasma muliebre is an asexual Hymenopteran wasp that is geographically and genetically isolated from all sexual relatives. D. muliebre attack larvae of the western cherry fruit fly (Rhagoletis indifferens), which in turn feed inside bitter cherry fruit (Prunus emarginata) in August and September. R. indifferens has recently colonized a new host plant with an earlier fruiting phenology (June/July), domesticated sweet cherries (P. avium), and D. muliebre has followed its host into this temporally earlier niche. We tested three hypotheses: 1) that all D. muliebre lineages originate from a single asexual ancestor; 2) that different D. muliebre lineages (as defined by unique mtDNA haplotypes) have differentiated on their ancestral host in an important life‐history trait, eclosion timing; and 3) that early‐eclosing lineages have preferentially colonized the new sweet cherry niche. We find that mitochondrial COI and microsatellite data provide strong support for a single ancestral origin for all lineages. Furthermore, COI sequencing revealed five mitochondrial haplotypes among D. muliebre, and individual wasps possessing one distinctive mitochondrial haplotype (haplotype II) eclosed as reproductive adults significantly earlier than wasps with all other haplotypes. In addition, this early‐eclosing lineage of D. muliebre is one of two lineages that have colonized the P. avium habitat, consistent with the preferential colonization hypothesis. These data suggest that D. muliebre has evolved adaptive phenotypic variation despite limited genetic variation, and that this variation has subsequently allowed an expansion of some wasps into a novel habitat. The D. muliebre system may allow for in‐depth study of adaptation and long‐term persistence of asexual taxa.     

Ultrastructure meets reproductive success: performance of a sphecid wasp is correlated with the fine structure of the flight-muscle mitochondria

Organisms show a remarkable inter-individual variation in reproductive success. The proximate causes of this variation are not well understood. We hypothesized that the ultrastructure of costly or complex tissues or organelles might affect reproductive performance. We tested this hypothesis in females of a sphecid wasp, the European beewolf, Philanthus triangulum (Hymenoptera, Sphecidae), that show considerable variation in reproductive success. The most critical component of reproduction in beewolf females is flying with paralysed honeybees, which more than double their weight. Because of the high energetic requirements for flight, we predicted that the ultrastructure of the flight-muscle mitochondria might influence female success. We determined the density of mitochondria and the density of the inner mitochondrial membranes (DIMM) of the flight muscles as well as age, body size and fat content. Only DIMM had a significant influence on female reproductive success, which might be mediated by an elevated adenosine triphosphate (ATP) supply. The variation in DIMM might result from differences in larval provisions or from an accumulation of mutations in the mitochondrial genome. Our results support the hypothesis that the organization of complex structures contributes to inter-individual variation in reproductive success.     

A learning-related variation in electroantennogram responses of a parasitic wasp

ABSTRACT Odour responses of parasitic insects, in search of their hosts, can change due to experience. Leptopilina heterotoma (Thomson) (Hymenoptera; Eucoilidae), a parasitic wasp of drosophilid larvae, is known to alter its preference for odours emanating from host food substrates through learning. These kinds of behavioural modifications in insects are assumed to be the result of complex processes in the brain. The results presented in this report, however, suggest that this learning-related behavioural variation is not restricted to brain processes but that it involves changes in sensitivity of olfactory receptor neurones.     

Rate of nutrient allocation to egg production in a parasitic wasp

Recent years have seen a marked increase in our awareness of the need to incorporate greater physiological realism into studies of parasitoid behaviour and population dynamics. Quantification of the number of eggs produced as a result of a host-feeding event, the host-feeding gain, is essential for predicting when a parasitoid should bypass an opportunity for current reproduction (i.e. laying eggs) in order to feed from the host and, thus, increase its chances for future reproduction (i.e. producing further eggs). Using radioactively labelled amino acids, one of the main constituents of insect haemolymph, we followed the incorporation of a known quantity of nutrients into each individual egg laid over a long period of time relative to the average life span of the parasitoid. Although the maximum incorporation of nutrients obtained by the female from a discrete feeding event occurs within a short period of time, a large proportion of nutrients are stored and used gradually for egg production throughout the life of the parasitoid. We therefore provide novel experimental evidence showing that feeding gain is not a discrete event in time occurring shortly after feeding, as has so far been assumed, but is instead spread throughout the parasitoid''s lifetime. This has important consequences for calculating the increase in lifetime fitness as a result of a feeding event, a common currency of models that aim to predict feeding and oviposition behaviour in parasitoids.     

Plant sexual systems correlated with morphological traits: Reflecting reproductive strategies of alpine plants

Plant sexual systems can be affected by various environmental factors, and thus show a range of morphological differences. The associations between plant sexual systems and morphology are unique, for example, the subnival belt of the Hengduan Mountains has harsh climatic conditions, but hosts remarkable biodiversity. However, these associations have received surprisingly little attention. We analyzed the associations in this region, and compared the pattern of sexual systems with that of other floras, and we also reviewed the breeding system of 36 alpine species. Information on plant sexual systems and these traits were mainly based on published information, herbarium collections, and field observations. The results showed that there was a high proportion of hermaphroditic species and a low proportion of dioecious and monoecious species. Hermaphroditism was strongly associated with showy floral displays, specialist entomophily, and perennial forbs. Dioecy was strongly associated with inconspicuous, pale-colored flowers, anemophily, and shrubs, whereas monoecy was strongly associated with inconspicuous, pale-colored flowers, generalist entomophily, and perennial grasses. Although no association was found between sexual systems and fruit type, plants with dry fruits (98.1%) were predominant in our study region. With respect to the plant breeding systems in the alpine region of southwest China, we predicted that pollination service might be particularly low; most of the plants lacked enough outcrossing pollinators. Our data revealed that 97.1% of the hermaphroditic species were self-compatible, and 88.2% of them showed autonomous or facilitated selfing. The plant sexual systems identified might serve to optimize the probability of pollination and thus ensure reproductive success.     

Cuticular hydrocarbons correlated with reproductive status in a queenless ant

Reproductive division of labour is regulated behaviourally in social insects lacking morphologically specialized castes. The directional nature of dominance interactions shows that recognition occurs, but little is known about its basis. In the queenless ant Dinoponera quadriceps, the top worker in the hierarchy (''alpha'') mates and produces offspring in each colony, while other workers remain virgin. Dominant ants frequently rub one antenna of subordinates against their own cuticle, and alpha and infertile nest-mates consistently differ in their relative proportions of the cuticular hydrocarbon 9-hentriacontene (9-C₃₁). The second-ranking ''beta'' occasionally lays unfertilized eggs and we show that she has less 9-C₃₁ than the alpha but more than infertile workers. To investigate further the link between 9-C₃₁ and ovarian activity, we experimentally removed alpha workers (n=11 individuals) and used solid-phase microextraction (SPME) with gas chromatography to measure changes in 9-C₃₁ on live beta workers which attained alpha status. The proportion of 9-C₃₁ on the replacement alpha increased significantly after six weeks, in parallel with her gain in fecundity. We discuss whether 9-C₃₁ provides honest information about egg-laying ability, enabling ants to recognize the different classes of nest-mates involved in reproductive conflicts. Such fertility cues could reliably underpin the antagonistic interactions occurring in insect societies.     

Characterization of a hymenoptaecin-like antimicrobial peptide in the parasitic wasp Nasonia vitripennis

Hymenoptaecin is a Hymenoptera insect-specific, glycine-rich antimicrobial peptide (AMP) found in non-parasitic bees. Here, we describe a unique hymenoptaecin-like gene (named nahymenoptaecin-1) in the parasitic wasp Nasonia vitripennis, which codes for a larger protein precursor with a carboxyl-terminal hymenoptaecin-like domain (HLD) similar to the bee hymenoptaecin. We recombinantly produced its full-length bioactive form as well as 1–33 and 34–98 fragments (named HLD-n and HLD-c, respectively). Recombinant HLD exhibited activity against Gram-negative and Gram-positive bacteria at micromolar concentrations. Compared to the full-length peptide, HLD-c possessed similar potency in inhibiting the growth of Stenotrophomonus but had a narrower antibacterial spectrum, whereas HLD-n only displayed weak effect on Stenotrophomonus, suggesting that HLD-n is a crucial determinant for bacterial target selectivity while HLD-c represents its active unit for the whole molecule. Circular dichroism analysis combined with ab initio structure prediction by Robetta indicated that HLD-n adopts a random coil conformation whereas glycine-rich HLD-c forms a loose β-sheet structure. Relative to bee hymenoptaecin, the upstream region of HLD contains two accurately repeated proline-rich AMP-like peptides instead of an acidic propeptide. Such difference could be a consequence of exon shuffling of autonomous modules after speciation.     

Functional response to host density in a parasitic wasp,with reference to population regulation

1. The functional response to, and preference for, the host density in a parasite were examined experimentally using an icheumon wasp, Exidechthis canescens, and its host Cadra cautella under controlled conditions.
2. Wasps were more active in host-searching at higher than lower host densities. Percent parasitism increased rapidly with initial increments in host density and then tended to increase more slowly at higher host densities. A sigmoid functional response curve is indicated, which implies that the parasite is able to control its host even at low densities.
3. Wasps actively selected areas of high host density in which to concentrate host-searching behavior.
4. Host-searching by E. canescens is stimulated by the odor of the host when present, and by food in which hosts have developed but have been removed.
5. Both the functional response and the host-density preference of the parasite are mediated by its host-searching behavior. This relationship is discussed in the context of population regulation.

     

Haemocyte changes in resistant and susceptible strains of D. melanogaster caused by virulent and avirulent strains of the parasitic wasp Leptopilina boulardi

Two strains of Drosophila melanogaster (resistant and susceptible) were parasitized by a virulent or avirulent strain of the parasitoid wasp Leptopilina boulardi. The success of encapsulation depends on both the genetic status of the host strain and the genetic status of the parasitoid strain: the immune cellular reaction (capsule) is observed only with the resistant strain-avirulent strain combination. The total numbers of host haemocytes increased in all 4 combinations, suggesting that an immune reaction was triggered in all hosts. Resistant host larvae infected with the virulent or avirulent strains of parasitoid wasp had slightly more haemocytes per mm(3) than did susceptible host larvae at the beginning of the reaction (less than 15 h post-parasitization). This difference disappeared later. Only the virulent parasitoid strain caused the production of a high percentage of altered lamellocytes (from a discoid shape to a bipolar shape), half the total number of lamellocytes are altered. This suggests that the alteration of lamellocyte shape alone is not sufficient to explain the lack of capsule formation seen in resistant hosts parasitized by the virulent strain. Lastly, there were very few altered lamellocytes in resistant or susceptible hosts parasitized by the avirulent parasitoid strain, two combinations in which no capsule was formed. As is now established for Drosophila-parasitoid interactions, virus-like particles contained in the long gland of the female wasp affect the morphology of the lamellocytes. The results presented here are further proof of the action (direct or indirect) of virus like particles of the virulent strain on lamellocytes.     

Seasonal variation in the reproductive strategy of the parasitic wasp Eulophus larvarum (Hymenoptera: Chalcidoidea: Eulophidae)

ABSTRACT. 1. Eulophus larvarum (L.) (Hymenoptera: Eulophidae) is a gregarious parasitoid of lepidopterous larvae feeding on broad-leaved trees. Normally there are two generations a year.
2. Sex ratio in the spring generation of larvae is female biased. The bias is probably due to local mate competition as progeny from one brood emerge from their pupae in close proximity to each other.
3. Sex ratio in the summer generation of larvae is near equality. Local mate competition is absent as individuals from the same brood become separated during the winter.
4. Variation in sex-ratio in the spring generation is consistent with a binomial model. Variation in sex ratio in the summer generation is much greater than expected from a binomial model and there is a large proportion of single-sex broods.
5. Two hypotheses are put forward to explain the summer generation pattern: virgin oviposition, and strong intersexual competition.
6. No differences in clutch size were found between the two generations.     

Two heparanase splicing variants with distinct properties are necessary in early Xenopus development

Heparanase is an endoglycosidase that cleaves heparan sulfate (HS) side chains from heparan sulfate proteoglycans (HSPGs) present in extracellular matrix and cell membranes. Although HSPGs have many functions during development, little is known of the role of the enzyme that degrades HS, heparanase. We cloned and characterized the expression of two heparanase splicing variants from Xenopus laevis and studied their function in early embryonic development. The heparanase gene (termed xHpa) spans over 15 kb and consists of at least 12 exons. The long heparanase (XHpaL) cDNA encodes a 531-amino acid protein, whereas the short splicing variant (XHpaS) results in a protein with the same open reading frame but missing 58 amino acids as a consequence of a skipped exon 4. Comparative studies of both isoforms using heterologous expression systems showed: 1) XHpaL is enzymatically active, whereas XHpaS is not; 2) XHpaL and XHpaS interact with heparin and HS; 3) both proteins traffic through the endoplasmic reticulum and Golgi apparatus, but XHpaL is secreted into the medium, whereas XHpaS remains associated with the membrane as a consequence of the loss of three glycosylation sites; 4) overexpression of XHpaS but not XHpaL increases cell adhesion of glioma cells to HS-coated surfaces; 5) XHpaL and XHpaS mRNA and protein levels vary as development progresses; 6) specific antisense knock-down of both XHpaL and XHpaS, but not XHpaL alone, results in failure of embryogenesis to proceed. Interestingly, rescue experiments suggest that the two heparanases regulate the same developmental processes, but via different mechanisms.     

Variants in Deleted in AZoospermia-Like (DAZL) are correlated with reproductive parameters in men and women

Qualitative and quantitative defects in human germ cell production that result in infertility are common and determined at least in part by genetic factors [Matzuk and Lamb, Nat Cell Biol 4(Suppl):s41–s49, 2002]. Yet, very few genes that are associated with germ cell defects in humans have been identified. In this study, we examined whether variants of the Deleted in AZoospermia-Like (DAZL) gene are associated with measures of germ cell production in three distinct populations of men and women. We identified 95 sequence variants in DAZL and further analyzed twelve single nucleotide polymorphisms (SNPs) that were present across ethnicities. We found that seven of the twelve SNPs were associated with at least one of the parameters studied (age at premature ovarian failure or menopause, total sperm count, or total motile sperm count). Surprisingly, many alleles exhibited opposing effects in men and women, which may be a result of different genetic requirements in male and female germ cells. Single SNP and haplotype analysis suggested that SNPs in the DAZL gene may act jointly to affect common reproductive characteristics in the human population. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users. Joyce Y. Tung and Mitchell P. Rosen have contributed equally     

Difference in carbon weight of the immature stages of two co-existing species of blackflies (Diptera: Simuliidae) with contrasting reproductive strategies

Two species of blackflies (Diptera: Simuliidae) co-exist at a lake-outlet site in Finland. The adult females of one of them, Simulium noelleri Fried., are able to produce eggs without taking a blood meal; females of the other species, S. truncalum Ldstr., must suck blood before they can mature eggs. Female pupae, pharate adults and adults of S. noelleri are markedly heavier in carbon weight than males of this species, and of males and females of S. truncatum. Females of the former species develop a fat body during the larval stadia.