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Phenoloxidase-like activities and the function of virus-like particles in ovaries of the parthenogenetic parasitoid Venturia canescens.

The ichneumonid endoparasitoid Venturia canescens successfully develops inside the hemocoel of another insect by using maternal protein secretions, including nucleic acid-free virus-like particles (VLPs), to manipulate host physiology. These VLPs consist of four major proteins, which are produced mainly in the calyx tissue and transferred into the host insect together with the egg. One of the protein-coding genes (vlp1), with similarities to phospholipid-hydroperoxide glutathione peroxidases (PHGPx), exists in allelic forms producing two protein variants with different protein properties. Here, we summarise observations indicating that oocytes and eggs are the source of reactive electrons, which potentially damage the lining and membranes of calyx tissues. We discuss the possible role of VLP1 in counteracting the damaging effects of oxidised phospholipids on membranes surrounding VLPs in the calyx lumen.

Animals↗

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.

Animals↗

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.

Animals↗

The genome sequence of an ichneumonid wasp, Venturia canescens (Gravenhorst, 1829) (Hymenoptera: Ichneumonidae).

We present a genome assembly from an individual female Venturia canescens (ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence has a total length of 299.96 megabases. Most of the assembly (97.25%) is scaffolded into 11 chromosomal pseudomolecules. The mitochondrial genome has also been assembled, with a length of 27.35 kilobases. Gene annotation of this assembly on Ensembl identified 14 281 protein-coding genes. This assembly was generated as part of the Darwin Tree of Life project, which produces reference genomes for eukaryotic species found in Britain and Ireland.

Hymenoptera↗

Microsatellite frequency and size variation in the parthenogenetic parasitic wasp Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae).

Nine genomic libraries of the parthenogenetic wasp Venturia canescens were screened for microsatellite loci. In contrast to other Hymenoptera (GT)n and not (CT)n, was the predominant repeat category found with 14 kb and 28 kb genomic DNA between loci, respectively. Mono- and trinucleotide microsatellites were rarer, occurring at frequencies between 231 kb and 589 kb of genome, whilst tetranucleotide repeats are scarce, with (ATTC)n and (CCGG)n loci occurring every 692 kb and 983 kb, respectively, and only one small imperfect (GATA)n locus and no (GACA)n loci were revealed. Over 70% of the dinucleotide, and all the trinucleotide microsatellites were small (less than eleven repeats), whilst 60% to 80% of loci were imperfect. Moreover, very few compound microsatellites and only a single association between different microsatellites were observed.

Animals↗

Lifetime egg maturation by host-deprived Venturia canescens.

The study investigated egg maturation and deposition by the endoparasitoid wasp Venturia canescens under conditions of host deprivation. Female V. canescens maintained without hosts began to deposit eggs onto the sides of the culture vessel on the day of eclosion. The maturation of additional eggs was not inhibited once the maximum oviduct egg load was reached but rather continued for the duration of the experiment (up to 39 days), at a rate of around 5.8% of the remaining unmatured eggs per day. Following host access, wasps matured additional eggs at an increased rate. Artificial damage to the ovipositor resulted in a reduced rate of egg maturation even though the oviducts were partly egg depleted, while damage to the auxiliary valvulae had no effect. These results suggest two conclusions. Under conditions of host deprivation, the rate at which eggs are matured is determined by the rate of synthesis of precursors by the fat body that in turn is modified by feedback from the ovipositor, induced by physical stimulation. Further, the discarding of eggs is due to the involuntary unidirectional movement of eggs down the oviduct, facilitated by the ongoing maturation of additional eggs.

Animals↗

Differential gene expression in two strains of the endoparasitic wasp Venturia canescens identified by cDNA-amplified fragment length polymorphism analysis.

In the endoparasitic wasp Venturia canescens (Hymenoptera: Ichneumonidae) two genetically distinct lines, which differ at the VLP1 gene locus, appear to coexist sympatrically in thelytokous as well as arrhenotokous field and laboratory strains. Both lines display quite distinct morphological, physiological and behavioural variations, such as different oviposition strategies, egg numbers and growth rates during early embryonic development. To examine whether more than one gene is affected in the two lines, we applied cDNA-amplified fragment length polymorphism analysis to examine patterns of gene expression in ovaries of both wasp lines. We show that a number of ovarian transcripts have an altered expression pattern in either line, which was further confirmed by virtual Northern blot analysis. Sequence analysis of the cDNA-amplified fragment length polymorphism fragments revealed that some of the respective genes are expected to be involved in regulation of protein degradation during stress responses and in signal perception/transduction. The full-length sequence of two transcription factors (a homeodomain containing protein and a zinc finger protein) differentially expressed in both lines was obtained by RACE-polymerase chain reaction and their putative role in regulating key developmental processes during embryogenesis is discussed.

Amino Acid Sequence↗

An empirical model of the sympatric coexistence of two strains of the endoparasitoid wasp Venturia canescens.

Recent research has demonstrated that a laboratory culture of the asexual solitary endoparasitoid wasp Venturia canescens Grav. (Hymenoptera: Ichneumonidae) contains two genetically and phenotypically distinct lines, coexisting on their host the flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae). The basis to the coexistence of the two lines appears to be differences in their reproductive success under single parasitism and superparasitism. Furthermore, examination of field-derived wasps from several locations has shown that the phenotypes displayed by the laboratory colonies also co-occur in field populations. Historically, the impossibility of showing that two species do not occupy separate niches has precluded any demonstration of sympatric coexistence in the field. Here we present the results of an iterative model that uses a range of experimental life history data to predict the stable composition of a mixed population of two lines displaying the laboratory phenotypes under different rates of superparasitism. The model predicts that sympatric coexistence of the two lines is possible when the overall rate of superparasitism is between 4 and 12% or greater. These values are within the rates reported for other solitary endoparasitoid wasp species in the field, and so demonstrate that the sympatric coexistence under natural conditions of two species that display the phenotypes observed in the laboratory lines is, in principle, possible.

Animals↗

Effects of lipophorin and 20-hydroxyecdysone on in vitro development of the larval endoparasitoid Venturia canescens (Hymenoptera: Ichneumonidae).

We previously reported that development of the embryos of the larval endoparasitoid Venturia canescens proceeded in the insect cell culture medium, MGM-450, and was promoted by the addition of a pupal extract from their host Galleria mellonella. The developmental promotion also was obtained by supplementing an equal amount of chicken egg yolk instead of the pupal extract to the medium. In this case, approximately 30% of the embryos developed into the second instar, but the value increased to more than 90% by treatment with 20-hydroxyecdysone. The medium supplemented with a G. mellonella pupal extract obtained by using Carlson's solution displayed growth-promoting ability, and in the extract, apolipophorin I was electrophoretically detected in large amounts. Both lipophorin purified from G. mellonella pupae and low density lipoprotein from chicken egg yolk acted as a growth-promoting substance for parasitoid development, although fetal bovine serum and 20-hydroxyecdysone were required as supplements to the medium for the expression of the ability. This indicated that lipophorin or lipophorin-transported lipids could act as a substance closely related to the growth-promoting factor(s) putatively involved in the host extract.

Journal Article↗

Cloning of a VLP-protein coding gene from a parasitoid wasp Venturia canescens.

Virus-like particles (VLPs) that are bound to the egg surface of a parasitic wasp Venturia canescens are void of nucleic acid and unable to infect host tissue but instead provide a passive protection against the host's immune recognition. To investigate evolutionary and functional properties of Venturia particles, we isolated cDNAs coding for a VLP-protein by screening an expression library of the wasp, using antibodies against purified VLPs. The corresponding coding DNA is present in the wasp genome as a single-copy gene.

Animals↗

Two coexisting lines of the endoparasitoid Venturia canescens show differences in reproductive success under conspecific superparasitism.

In a laboratory colony of the endoparasitic wasp Venturia canescens Grav. (Hymenoptera: Ichneumonidae), two genetically distinct lines (RP, RM) appear to coexist sympatrically. The two lines display pronounced differences in ovarian morphology, parasitism behaviour and number of offspring produced under competing superparasitism. Since V. canescens is a solitary endoparasitoid, larvae inside superparasitised hosts must compete for host possession. We examined the outcome of conspecific superparasitism between the wasp lines with different time intervals between ovipositions. The results showed that the competitive abilities of the two lines were not symmetrical. Further, the RM-line won a significantly higher fraction (around 60%) of the overall contests. Dissection of parasitoid larvae from their hosts indicated that most contests between competing larvae had occurred within the first 24 h of the eggs hatching, suggesting the advantage of the RM-line relates to physical combat. It was previously thought that the coexistence of the two lines was exclusively due to maternal effects. The results of this study indicate for the first time that these differences are based on phenotypic variations in both the larval offspring and the mother.

Age Factors↗

Automictic parthenogenesis in the parasitoid Venturia canescens (Hymenoptera: Ichneumonidae) revisited.

Both arrhenotokous and thelytokous reproduction are known to occur in the parasitoid wasp Venturia canescens. The cytological mechanism of thelytoky was previously reported to involve the formation of a restitution metaphase after the reduction division, but the exact nature of the subsequent divisions, whether reductional or equational, remained unclear. We reinvestigated the cytological mechanisms in a thelytokous strain collected in France. Our observations confirm previous results, but an equational and not a reduction division was observed after restitution. This type of reproduction can be classified as central fusion automictic parthenogenesis. In two arrhenotokous strains the normal pattern of oogenesis and syngamy of Hymenoptera was observed. In addition, we used PCR amplification to show that thelytoky in V. canescens is not caused by Wolbachia bacteria. The results are discussed in relation to maintenance of heterozygosity and female sex.

Animals↗

Nuclear magnetic resonance microscopy of the development of the parasitoid wasp Venturia canescens within its host moth Plodia interpunctella.

Nuclear magnetic resonance microscopy was used to image the parasitoid wasp Venturia canescens (Hymenoptera: Ichneumonidae) within larval and pupal instars of its host, the Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae). The images were obtained using gradient-echo and chemical shift selective pulse sequences and clearly showed the location and shapes of the parasitoid as it developed from the L1 larva to a pupal stage within the host. The digestive, nervous, and tracheal systems of the host were identified and changes were observed as the host underwent metamorphosis. Destruction of the host tissues by the parasitoid was visible. It was found that the parasitoid first ate the fat body and digestive system of the host, allowing the host to continue to grow, and only progressed to the vital organs when its own development had neared pupation.

Animals↗

The development of the endoparasitoid wasp Venturia canescens in superparasitised Ephestia kuehniella.

Using a molecular marker that allows the differentiation of two strains of the solitary endoparasitoid wasp Venturia canescens, the study investigated the influence of host mass and the time interval between ovipositions on the survival and development of larvae from both the first and second laid eggs in superparasitised Ephestia kuehniella. As the time interval between ovipositions increased both overall and superparasitism success decreased, however, time between, and order of, ovipositions had little effect on other developmental parameters. Adult size increased with host mass under both parasitism and superparasitism, while host mortality decreased with host mass under superparasitism. In addition, wasps emerging from superparasitised hosts were larger than wasps from parasitised hosts. The results confirm that for V. canescens on the host E. kuehniella both self- and conspecific-superparasitism will be an adaptive strategy when hosts are the limiting factor.

Adaptation, Physiological↗

Gene expression changes in Ephestia kuehniella caterpillars after parasitization by the endoparasitic wasp Venturia canescens analyzed through cDNA-AFLPs.

To understand the transcriptional response of Ephestia kuehniella caterpillars to parasitization by the endoparasitic wasp Venturia canescens, we examined patterns of gene expression in parasitized and unparasitized host caterpillars using the cDNA-AFLP (cDNA-amplified fragment length polymorphism) technique. Among the approximately 1860 cDNA-fragments detected by using 88 different AFLP primer combinations, we identified 13 transcripts in parasitized caterpillars that showed a suppressed expression pattern with cDNA-AFLP results being confirmed by virtual Northern blot or RT-PCR analysis. Apparently, two different wasp genotypes (RP, RM) seem to differ in their ability to regulate host physiology with regard to suppression of certain genes. Sequence analysis of the cDNA fragments revealed that some of the respective genes are likely to be involved in energy metabolism, protein synthesis, or in signal transduction. In addition, one sequence corresponds to a gene encoding the lepidopteran silk protein fibroin. The putative function of these genes in the interactions between endoparasitoids and their host insects is discussed.

Animals↗

The development of the endoparasitoid Venturia canescens in Bt-tolerant, immune induced larvae of the flour moth Ephestia kuehniella.

We examined the survival and development of the endoparasitoid Venturia canescens in a Bt-tolerant laboratory strain of the flour moth Ephestia kuehniella, in which Bt-tolerance has been shown to be associated with an inducible elevation of the insect's immune response. The results indicate the elevated immune status associated with Bt-tolerance does not confer cross-protection against parasitism by V. canescens. No significant difference was observed in the rate of emergent wasps from parasitised Bt-tolerant and Bt-susceptible hosts. In addition, wasps from Bt-tolerant hosts had longer development times and were larger than wasps from Bt-susceptible hosts.

Animals↗

Life tables of Venturia canescens (Hymenoptera: Ichneumonidae) parasitizing the Mediterranean flour moth (Lepidoptera: Pyralidae).

Effects of temperature, adult feeding, and host instar on life table parameters of Venturia canescens Gravenhorst (Hymenoptera: Ichneumonidae) parasitizing larvae of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) were studied in the laboratory. Experimental adults lived under various regimes of temperature (15, 20, 25, and 30 degrees C), food supply (with or without access to honey), and host instar (second, third, fourth, and fifth). Temperature increase resulted in higher values of the intrinsic rate of natural increase (r(m)), the net reproductive rate (R(o)), the finite capacity of increase (lambda), and the gross reproductive rate (GRR), whereas it was followed by decrease of the mean generation time (G) and the doubling time (DT) values. Feeding on honey caused remarkable increase of r(m), R(o), and GRR, whereas r(m) and lambda reached their maximum when full-grown hosts (fifth instar) were parasitized. This is the first time life table parameters of V. canescens have been studied. The findings of the current study are discussed on the basis of improving V. canescens performance as a biological agent against moth pests of stored products.

Animals↗

Growth-promoting effects of ecdysteroids and juvenile hormone on in vitro development of the larval endoparasitoid, Venturia canescens (Hymenoptera: Ichneumonidae).

We previously reported that lipophorin, fetal bovine serum (FBS), and 20-hydroxyecdysone (20-HE) are essential for the development of the larval endoparasitoid Venturia canescens larvae in vitro. The present study was undertaken to determine the optimal concentrations of those three substances in the MGM-450 medium, and to examine the hormonal effects of ecdysteroids and juvenile hormone (JH) on the development of the parasitoid larvae in vitro. When the culture was started with embryos at the post-germband stage, concentrations of 3 mg/ml of lipophorin and 20% of FBS were most suitable for the development of the parasitoid. The growth-promoting effect of 20-HE increased in a concentration-dependent manner and peaked at a concentration of 1 &mgr;g/ml. Excess concentration led to malformations of the larvae. Three other ecdysteroids, ecdysone, 2-deoxy-20-hydroxyecdysone, and polypodine B had the same effect, although their activity was lower than that of 20-HE. Cholesterol had no effect; most larvae failed to develop. When the medium was supplemented with JH, the duration of the developmental period was significantly shortened, but this hormone was not found to be essential.

Journal Article↗