Conserving genetic variability of a wild insect population under laboratory conditions

Drosophila melanogaster (Meigen) was used to test the power of isofemale lines in preserving genetic variability. We performed experiments in two ways. One series consisted of measuring the genetic variability for three enzymatic loci in 32 isofemale lines, in the first and 23^rd generations of culture. In the second series, we tested the capacity of the larvae to eliminate a parasitoid by encapsulation after eight years of laboratory breeding. In general, individual isofemale lines appeared to change during the 23 generations of the study, but the global frequency of these alleles among the 32 isofemale lines stayed relatively unchanged. The only rare allele observed was also conserved. Changes in allozyme frequencies at any one locus were independent of those at other loci. Genetic variation of a biological trait, the capacity of the larvae to encapsulate a parasitoid, also changed, but it could be restored to a level close to that of the starting population by mass hybridizing together individuals of each line.

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Résumé D. melanogaster (Meigen) a été utilisé pour tester la capacité des lignées isofemelles à conserver la variabilité génétique d'une population naturelle. Deux types d'expériences ont été réalisées. L'une a consisté à déterminer la variabilité génétique de 3 locus enzymatiques pour 32 lignées isofemelles à la première et à la 23^ème génération d'élevage au laboratoire. L'autre a consisté à tester la capacité des larves à éliminer un parasito?de par le processus d'encapsulation après 8 années d'élevage au laboratoire. D'une fa?on générale, certaines lignées isofemelles perdent de la variabilité durant les 23 générations de l'étude. Mais la fréquence globale des allèles reste inchangée si l'on considère l'ensemble des 32 lignées. Le seul allèle rare observé a également été conservé. Les modifications des fréquences allèliques à chacun des locus ont lieu de fa?on indépendante les unes des autres. La variabilité génétique d'un caractère biologique, la capacité des larves à encapsuler le parasito?de, a également varié, mais elle a pu être restaurée à un niveau proche de la population initiale en rassemblant plusieurs individus de chacune des lignées.

     

Utilization and tolerance of ethanol, acetic acid and acetaldehyde vapour by Asobara persimilis, a parasitoid of Drosophila

The tolerance and utilization of ethanol, acetic acid and acetaldehyde vapour was investigated in Asobara persimilis (Hymenoptera: Braconidae), a parasitoid of Drosophila. No significant utilization of ethanol or acetaldehyde occurred at low concentrations (< 1.5% and 0.1% respectively), however both female and male longevity was increased at concentrations of 1.0 and 1.5% acetic acid. All substances were toxic at higher concentrations, but there was sexual dimorphism in that females survived significantly longer than males.

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Résumé L'étude a porté sur la tolérance et l'utilisation de vapeurs d'éthanol, d'acide acétique et d'acétaldéhyde par Asobara persimilis (Hym. Braconidae), parasitoïde de Drosophila récemment découvert en Australie. Aux faibles concentrations, il n'y a pas d'utilisation significative d'éthanol et d'acide (respectivement moins de 1,5% et 0,1%), cependant la longévité des mâles et des femelles ont augmenté avec les concentrations d'acide acétique de 1,0 et 1,5%. Toutes ces substances sont toxiques à plus forte concentrations, bien qu'il y ait un dimorphisme sexuel et que les femelles survivent significativement plus longtemps que les mâles. La tolérance des braconides est inférieure à celle de leurs hôtes, les Drosophiles cosmopolites et endémiques à l'Australie. Ceci peut faire que ces parasites limitent l'exploitation de leurs hôtes aux habitats avec une faible concentration de produits de fermentation.

     

Egg-laying experience and acceptance of parasitized hosts by the parasitoid,Leptopilina heterotoma (Hymenoptera: Eucoilidae)

The influence of egg-laying experience on the response of females of the eucoilid parasitoid,Leptopilina heterotoma, to parasitized and unparasitizedDrosophila melanogaster host larvae was examined under more controlled conditions than those used in past studies. In laboratory assays, we precisely manipulated both the number of eggs laid by females and the kind of larvae (parasitized versus unparasitized) in which the eggs were laid. We found that the tendency to avoid laying eggs in parasitized hosts depended markedly on whether or not eggs had been laid previously, but depended little on whether those eggs had been laid in parasitized or unparasitized hosts. The observed effect of general egg-laying experience on avoidance of parasitized hosts may reflect responses to either changes in the wasp's internal state (perhaps, changes in egg load) or changes in the wasp's neural representation of the external environment (such as those presumed to occur during learning). In light of these results, we offer a tentative reinterpretation of several earlier studies.     

Larval parasitoid uses aggregation pheromone of adult hosts in foraging behaviour: a solution to the reliability-detectability problem

Parasitoids that forage for herbivorous hosts by using infochemicals may have a problem concerning the reliability and detectability of these stimuli: host stimuli are highly reliable but not very detectable at a distance, while stimuli from the host's food are very detectable but generally not very reliable in indicating host presence. One solution to this problem is to learn to link highly detectable stimuli to reliable but not very detectable stimuli. Ample knowledge is available on how associative learning aids foraging parasitoids in the location of suitable microhabitats. However, in this paper we report on another solution to the reliability-detectability problem and present evidence for an essential, but as yet overlooked, aspect of Drosophila parasitoid ecology. For the first time it is shown that a parasitoid of Drosophila larvae spies on the communication system of adult Drosophila flies to locate potential host sites: naive parasitoids strongly respond to a volatile aggregation pheromone that is deposited in the oviposition site by recently mated female flies. Thus, the parasitoids resort to using highly detectable information from a host stage different from the one under attack (i.e. infochemical detour). The function and ecological implications of these findings are discussed.     

Specific expression of the Drosophila midline-jumper retro-transposon in embryonic CNS midline cells

Here we describe of a novel Drosophila LTR-type retrotransposon that is expressed in the embryonic CNS midline glia and in the embryonic germ cells. The element is related to the gypsy and burdock retrotransposons and was termed midline-jumper. In addition to cDNA clones generated from internal retrotransposon sequences, we have identified one cDNA clone that appears to reflect a transposition event, indicating that the midline-jumper retrotransposon is not only transcribed but also able to transpose during Drosophila development.     

Phenotypic suppression of the Drosophila mitochondrial disease-like mutant tko by duplication of the mutant gene in its natural chromosomal context

A mutation in the Drosophila gene technical knockout (tko^25t), encoding mitoribosomal protein S12, phenocopies human mitochondrial disease. We isolated three spontaneous X-dominant suppressors of tko^25t (designated Weeble), exhibiting almost wild-type phenotype and containing overlapping segmental duplications including the mutant allele, plus a second mitoribosomal protein gene, mRpL14. Ectopic, expressed copies of tko^25t and mRpL14 conferred no phenotypic suppression. When placed over a null allele of tko, Weeble retained the mutant phenotype, even in the presence of additional transgenic copies of tko^25t. Increased mutant gene dosage can thus compensate the mutant phenotype, but only when located in its normal chromosomal context.     

Response of the egg parasitoid Trichogramma pretiosum to the sex pheromone of its host Heliothis zea

This paper presents results of olfactometer experiments with the egg parasitoid Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) and its host the corn earworm moth, Heliothis zea (Boddie) (Lepidoptera: Noctuidae). The sex pheromone of the host significantly reduced the total number of border crossings between odour fields in the olfactometer. Also, female parasitoids made significantly more visits to the calling moth odour field than to the opposite control field in the olfactometer. Further, the wasps spent significantly more time in the olfactometer field containing the sex pheromone released by calling virgin moths, than in control fields. If non-calling virgin moths were used as odour source, the response was reversed and wasps were repelled by the odour of the moths, and the numbers of visits were evenly distributed over the four flow fields. These results are discussed in the context of foraging ecology of egg parasitoids.

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Résumé Des expériences menées en olfactométrie avec le parasite oophage Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) et son hôte, Heliothis zea (Boddie) (Lepidoptera: Noctuidae) donnent les résultats suivants. La présence de la phéromone sexuelle de l'hôte réduit de façon significative le nombre de passages entre les champs odorisés. De mème, les parasites visitent plus fréquemment le champ qui dispense l'odeur de la femelle en appel que le champ témoin placé à l'opposé. Par ailleurs, le temps passé dans le champ qui contient la phéromone sexuelle émise par les femelles vierges en appel est significativement supérieur à la durée de visite des champs témoins. Si l'on utilise comme source d'odeur des femelles vierges qui ne sont pas en appel, la réponse est inversée, les parasites sont alors repoussés par l'odeur de ces papillons et le nombre de visites est distribué de façon aléatoire entre les quatre champs. Ces résultats sont discutés dans le contexte de l'écologie du comportement de recherche chez les parasites oophages.

     

Cell proliferation, survival, and death in the Drosophila eye

The Drosophila retina has a precise repeating structure based on the unit eye, or ommatidium. This review summarizes studies of the cell proliferation and survival episodes that affect the number of cells available to make each ommatidium. Late in larval development, as differentiation and patterning begin, the retinal epithelium exhibits striking regulation of the cell cycle including a transient G1 arrest of all cells, followed by a ‘Second Mitotic Wave’ cell cycle that is regulated at the G2/M transition by local intercellular signals. Reiterated episodes of cell death also contribute to precise regulation of retinal cell number. The EGF receptor homolog has multiple roles in retinal proliferation and survival.     

Uscana lariophaga, West-African egg parasitoid of the cowpea bruchid beetleCallosobruchus maculatus: photoperiod, parasitization and eclosion interactions

The diurnal pattern of parasitization and eclosion of the trichogrammatidUscana lariophaga Steffan, egg parasitoid of the cowpea bruchidCallosobruchus maculatus (Fabricius) was studied for L12:D12 photoperiods. The wasp parasitized eggs throughout each 24-h period with about 70% of the parasitization taking place during the first 12 h regardless whether it was the photophase or the scotophase. More female progeny was produced when the first 12-h period was photophase instead of scotophase. Eclosion of wasps took place over a period of 4 days and occurred during the second half of the scotophase and the first half of the photophase. The number of wasps eclosed during the photophase was similar to that eclosed during the scotophase. Average development time was 8.9 days at 30°C. Male development was completed 6 to 8 h before the females, hence a higher percentage of females emerged in the later eclosion peaks. During scotophases more females eclosed than during photophases. Although the results indicate that the wasp is able to perform under dark storage conditions the effect of permanent low light intensity remains to be studied.     

Host-age discrimination during host location byCotesia glomerata, a larval parasitoid ofPieris brassicae

Some parasitoids are restricted with respect to the host stage that they attack and even to a certain age within a stage. In this paper we investigate whether the parasitoidCotesia glomerata can discriminate between old and young caterpillar instars of its host,Pieris brassicae, before contacting these hosts, since contacts with older instars are very risky with a chance of being killed, due to the aggressive defensive behaviour of the caterpillars. Flight chamber dual choice tests showed that volatile chemicals emitted by Brussels sprouts plants (Brassica oleracea var. gemmifera) after feeding damage by 1st and 5th larval instars are equally attractive to the wasps. Simulated herbivore damage by 2nd and 5th larval instars, obtained by treating mechanically damaged leaves with carterpillar regurgitant, was also equally attractive, even when the wasps were exposed to repeated experience on different larval instars to increase their discriminatory ability. In contrast, single choice contact bioassays showed that the time spent searching on a leaf with feeding damage of 1st instar larvae was significantly longer than the time spent on 5th instar feeding damage or on mechanically damaged leaves. Both flight and contact bioassays did not show any effect of egg-related infochemicals. The results demonstrate thatC. glomerata can discriminate between young and old larval instars ofP. brassicae, without contacting the caterpillars. This is not done through volatile herbivore-induced synomones but through cues that are contacted after arrival at a caterpillar-infested leaf.     

Compatible invasion of a phylogenetically distant host embryo by a hymenopteran parasitoid embryo

Embryonic invasion into the tissue of genetically different organisms has been known only in mother-embryo interactions of viviparous organisms. Hence, embryonic invasions have been thought to occur only within the same or closely related species. For endoparasitic Hymenoptera, which are oviposited in their host egg but complete their development in the later stages, entry into the host embryo is essential. To date, the entry of these parasitoids is known to be accomplished by either egg deposition directly into the embryo or by the newly hatched larva boring into the embryo. However, Copidosoma floridanum is a polyembryonic parasitoid whose development is characterized by a prolonged embryonic stage, and which lacks a larval form during its host embryogenesis. We have analyzed the behavior and fate of C. floridanum embryos co-cultured with their host embryo in vitro. Here, we show that the morula-stage embryo of C. floridanum actively invades the host embryo. Histological analyses have demonstrated that C. floridanum embryonic invasion is associated with adherent junction to host cells rather than causing an obvious wound on the host cells. These findings provide a novel case of embryonic invasion into a phylogenetically distant host embryo, ensuring cellular compatibility with host tissues.     

Functional interactions between the gene tetanic and the Shaker gene complex of Drosophila

Different phenotypes associated with the tetanic (tta) mutation such as appendage contraction, maternal effect and low viability and fertility are enhanced by one extra dose of the Shaker gene complex (ShC). The tta mutation is lethal with two extra doses of ShC. In addition, tta embryos have a defective nervous system. In this paper, I analyse the interaction between tta and ShC to gain insight into their relationship. Aneuploid analysis suggests that the lethality is due to an interaction of the tta mutation with the maternal effect (ME) region of this gene complex. Mutations in the ME region of ShC partially suppress this interaction. Trans-heterozygous combinations of MEI[l(1)305] and MEIII [l(1)459] mutations causes dominant lethality in a tta background. Trans-heterozygous combinations of an MEII [l(1)1359] mutation with the cited MEI and MEIII mutations are lethal in a tta background. Double mutant combinations and gene dosage experiments, suggest that tta also interacts with the viable (V) region of ShC. These specific genetic interactions indicate that tta and the ME and V regions of ShC are functionally related. These results, together with the previous electrophysiological, molecular and biochemical studies on these mutants suggest an interaction at the protein level. Thus, in the case of the V region, the tta gene product may modulate the activity of the K⁺ channels encoded in this region. Furthermore, the extreme dosage sensitivity of the interaction between tta and ShC suggests a stoichiometric requirement for the different gene products involved, which might be physically associated and form heteromultimers.     

Host-acceptance behaviour of Alloxysta pleuralis, a cynipoid hyperparasitoid of an aphidiid parasitoid Trioxys indicus on aphids

Alloxysta pleuralis parasitizes the larvae of Trioxys indicus, a parasitoid of Aphis craccivora. The pattern of host-searching of A. pleuralis was random regarding the host-stage. Earlier second instar to early fourth instar larvae of T. indicus within the haemocoel of the living aphid were found susceptible for attack, however, the third instar larvae were more parasitized. The rate of oviposition was very low (8.5 eggs/h). The number of pricks into the aphid was significantly more for the aphids that contained susceptible stages of the potential host. The occurrence of superhyperparasitism was lower when the hyperparasitoids were offered a choice of all host stages than were offered no-choice.

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Résumé A. pleuralis est un hyperparasitoïde endophage de Trioxys indicus, parasitoïde d'Aphis craccivora, puceron de Cajanus cajan. Bien que les larves de T. indicus soient susceptibles d'être attaquées du début du second stade au début du quatrième stade, celles du troisième stade sont nettement préférées. L'absence de fifférence entre les fréquences de visites et de contacts avec les hôtes de différents stades a montré que le comportement de prospection de A. pleuralis s'effectue au hasard. Cependant, la fréquence des piqûres/hôte est significativement supérieure chez lez hôtes préférés, ce qui donne une indication sur le fait que A. pleuralis pique un puceron pour rechercher un hôte potentiel. Il est possible que la femelle perçoive la présence de l'hôte dans le puceron en décelant avec les récepteurs sensoiriels de sa tarière les changements physiologiques caractéristiques du puceron parasité, et qu'elle les utilise pour piquer à répétition le puceron jusqu'à ce qu'elle trouve l'hôte potentiel. Le superhyperparasitisme était relativement faible et plus fréquent dans les expériences où l'hyperparasite n'avait pas de choix.

     

The biology of Bracon celer as a parasitoid of the olive fruit fly

A series of laboratory experiments was conducted on a colony of Bracon celer Szépligeti (Hymenoptera: Braconidae) reared on the olive fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae). Female B. celer preferentially probe and oviposit into olives containing late third-instar fly larvae. The parasitoid develops as a solitary, ectoparasitic idiobiont. Mean development time (oviposition to adult eclosion) at 22 °C was, for females, 36±1 (SE) days, and for males, 34±1 days. The mean longevity of adult female wasps when provided honey and water was significantly greater than when they were provided water alone, or nothing. The females produced an average of 9.7±7.2 progeny during their lifetimes, but production levels in the insectary colony suggested that this level of fecundity was artificially low and could be improved. The discrepancy may be a consequence of constraints on oviposition behavior imposed by the experimental design. The results are discussed with respect to insectary production methods and the potential use of B. celer as a biological control agent for olive fly in California.     

Responses of a generalist and a specialist parasitoid (Hymenoptera: Eucoilidae) to Drosophilid larval kairomones

Foraging parasitoids are thought to need more specific information than generalists on the presence, identity, availability, and suitability of their insect host species. In the present paper, we compare responses to host kairomones by two phylogenetically related parasitoid species that attack Drosophilidae and that differ in the width of their host range. As predicted, the behavioral response of the parasitoids to host kairomones reflected their difference in host range. The response of the specialist parasitoid Leptopilina boulardiwas restricted to contact kairomones from its natural hosts and one closely related species. In contrast, the generalist parasitoid Leptopilina heterotomaresponded to contact kairomones of a variety of Drosophilidae species.     

In vivo modification of Na,K-ATPase activity in Drosophila

We have constructed and characterized transgenic Drosophila lines with modified Na⁺,K⁺-ATPase activity. Using a temperature dependent promoter from the hsp70 gene to drive expression of wild-type α subunit cDNA, we can conditionally rescue bang-sensitive paralysis and ouabain sensitivity of a Drosophila Na⁺,K⁺-ATPase α subunit hypomorphic mutant, 2206. In contrast, a mutant α subunit (α^D369N) leads to increased bang-sensitive paralysis and ouabain sensitivity. We can also generate temperature dependent phenotypes in wild-type Drosophila using the same hsp70 controlled α transgenes. Ouabain sensitivity was as expected, however, both bang sensitive paralysis or locomotor phenotypes became more severe regardless of the type of α subunit transgene. Using the Gal4-UAS system we have limited expression of α transgenes to cell types that normally express a particular Drosophila Na⁺,K⁺-ATPase β (Nervana) subunit isoform (Nrv1 or 2). The Nrv1-Gal4 driver results in lethality while the Nrv2-Gal4 driver shows reduced viability, locomotor function and uncontrolled wing beating. These transgenic lines will be useful for disrupting function in a broad range of cell types.     

Probing the Drosophila retinal determination gene network in Tribolium (II): The Pax6 genes eyeless and twin of eyeless

The Pax6 genes eyeless (ey) and twin of eyeless (toy) are upstream regulators in the retinal determination gene network (RDGN), which instructs the formation of the adult eye primordium in Drosophila. Most animals possess a singleton Pax6 ortholog, but the dependence of eye development on Pax6 is widely conserved. A rare exception is given by the larval eyes of Drosophila, which develop independently of ey and toy. To obtain insight into the origin of differential larval and adult eye regulation, we studied the function of toy and ey in the red flour beetle Tribolium castaneum. We find that single and combinatorial knockdown of toy and ey affect larval eye development strongly but adult eye development only mildly in this primitive hemimetabolous species. Compound eye-loss, however, was provoked when ey and toy were RNAi-silenced in combination with the early retinal gene dachshund (dac). We propose that these data reflect a role of Pax6 during regional specification in the developing head and that the subsequent maintenance and growth of the adult eye primordium is regulated partly by redundant and partly by specific functions of toy, ey and dac in Tribolium. The results from embryonic knockdown and comparative protein sequence analysis lead us further to conclude that Tribolium represents an ancestral state of redundant control by ey and toy.