Frédéric Fleury

Removing symbiotic Wolbachia bacteria specifically inhibits oogenesis in a parasitic wasp

Wolbachia are bacteria that live in the cells of various invertebrate species to which they cause a wide range of effects on physiology and reproduction. We investigated the effect of Wolbachia infection in the parasitic wasp, Asobara tabida Nees (Hymenoptera, Braconidae). In the 13 populations tested, all individuals proved to be infected by Wolbachia. The removal of Wolbachia by antibiotic treatment had a totally unexpected effect—aposymbiotic female wasps were completely incapable of producing mature oocytes and therefore could not reproduce. In contrast, oogenesis was not affected in treated Asobara citri, a closely related species that does not harbor Wolbachia. No difference between natural symbiotic and cured individuals was found for other adult traits including male fertility, locomotor activity, and size, indicating that the effect on oogenesis is highly specific. We argue that indirect effects of the treatments used in our study (antibiotic toxicity or production of toxic agents) are very unlikely to explain the sterility of females, and we present results showing a direct relationship between oocyte production and Wolbachia density in females. We conclude that Wolbachia is necessary for oogenesis in these A. tabida strains, and this association would seem to be the first example of a transition from facultative to obligatory symbiosis in arthropod–Wolbachia associations.

The Transmission Efficiency of Tomato Yellow Leaf Curl Virus by the Whitefly Bemisia tabaci Is Correlated with the Presence of a Specific Symbiotic Bacterium Species

ABSTRACT Tomato yellow leaf curl virus (TYLCV) (Geminiviridae: Begomovirus) is exclusively vectored by the whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). TYLCV transmission depends upon a 63-kDa GroEL protein produced by the vectors endosymbiotic bacteria. B. tabaci is a species complex comprising several genetically distinct biotypes that show different secondary-symbiont fauna. In Israel, the B biotype harbors Hamiltonella, and the Q biotype harbors Wolbachia and Arsenophonus. Both biotypes harbor Rickettsia and Portiera (the obligatory primary symbionts). The aim of this study was to determine which B. tabaci symbionts are involved in TYLCV transmission using B. tabaci populations collected in Israel. Virus transmission assays by B. tabaci showed that the B biotype efficiently transmits the virus, while the Q biotype scarcely transmits it. Yeast two-hybrid and protein pulldown assays showed that while the GroEL protein produced by Hamiltonella interacts with TYLCV coat protein, GroEL produced by Rickettsia and Portiera does not. To assess the role of Wolbachia and Arsenophonus GroEL proteins (GroELs), we used an immune capture PCR (IC-PCR) assay, employing in vivo- and in vitro-synthesized GroEL proteins from all symbionts and whitefly artificial feeding through membranes. Interaction between GroEL and TYLCV was found to occur in the B biotype, but not in the Q biotype. This assay further showed that release of virions protected by GroEL occurs adjacent to the primary salivary glands. Taken together, the GroEL protein produced by Hamiltonella (present in the B biotype, but absent in the Q biotype) facilitates TYLCV transmission. The other symbionts from both biotypes do not seem to be involved in transmission of this virus.

Endosymbiont metacommunities, mtDNA diversity and the evolution of the Bemisia tabaci (Hemiptera: Aleyrodidae) species complex

Bemisia tabaci, an invasive pest that causes crop damage worldwide, is a highly differentiated species complex, divided into biotypes that have mainly been defined based on mitochondrial DNA sequences. Although endosymbionts can potentially induce population differentiation, specialization and indirect selection on mtDNA, studies have largely ignored these influential passengers in B. tabaci, despite as many as seven bacterial endosymbionts have been identified. Here, we investigate the composition of the whole bacterial community in worldwide populations of B. tabaci, together with host genetic differentiation, focusing on the invasive B and Q biotypes. Among 653 individuals studied, more than 95% of them harbour at least one secondary endosymbiont, and multiple infections are very common. In addition, sequence analyses reveal a very high diversity of facultative endosymbionts in B. tabaci, with some bacterial genus being represented by more than one strain. In the B and Q biotypes, nine different strains of bacteria have been identified. The mtDNA‐based phylogeny of B. tabaci also reveals a very high nucleotide diversity that partitions the two ITS clades (B and Q) into six CO1 genetic groups. Each genetic group is in linkage disequilibrium with a specific combination of endosymbionts. All together, our results demonstrate the rapid dynamics of the bacterial endosymbiont–host associations at a small evolutionary scale, questioning the role of endosymbiotic communities in the evolution of the Bemisia tabaci species complex and strengthening the need to develop a metacommunity theory of inherited endosymbionts.

Inherited intracellular ecosystem: symbiotic bacteria share bacteriocytes in whiteflies

Symbiotic relationships with bacteria are common within the Arthropoda, with interactions that substantially influence the biology of both partners. The symbionts’ spatial distribution is essential for understanding key aspects of this relationship, such as bacterial transmission, phenotype, and dynamics. In this study, fluorescence in situ hybridization was used to localize five secondary symbionts from various populations and biotypes of the sweet potato whitefly Bemisia tabaci: Hamiltonella, Arsenophonus, Cardinium, Wolbachia, and Rickettsia. All five symbionts were found to be located with the primary symbiont Portiera inside the bacteriocytes — cells specifically modified to house bacteria — but within these cells, they occupied various niches. The intrabacteriocyte distribution pattern of Rickettsia differed from what has been described previously. Cardinium and Wolbachia were found in other host tissues as well. Because all symbionts share the same cell, bacteriocytes in B. tabaci represent a unique intracellular ecosystem. This phenomenon may be a result of the direct enclosure of the bacteriocyte in the egg during oogenesis, providing a useful mechanism for efficient vertical transmission by “hitching a ride” with Portiera. On the other hand, cohabitation in the same cell provides ample opportunities for interactions among symbionts that can either facilitate (cooperation) or limit (warfare) symbiotic existence.—Gottlieb, Y., Ghanim, M., Gueguen, G., Kontsedalov, S., Vavre, F., Fleury, F., Zchori‐Fein, E. Inherited intracellular ecosystem: symbiotic bacteria share bacteriocytes in whiteflies. FASEB J. 22, 2591–2599 (2008)

EVIDENCE FOR FEMALE MORTALITY IN WOLBACHIA‐MEDIATED CYTOPLASMIC INCOMPATIBILITY IN HAPLODIPLOID INSECTS: EPIDEMIOLOGIC AND EVOLUTIONARY CONSEQUENCES

Abstract.— Until now, only two Wolbachia‐mediated cytoplasmic incompatibility (CI) types have been described in haplodiploid species, the first in Nasonia (Insect) and the second in Tetranychus (Acari). They both induce a malebiased sex ratio in the incompatible cross. In Nasonia, CI does not reduce fertility since incompatible eggs develop as haploid males, whereas in Tetranychus CI leads to a partial mortality of incompatible eggs, thus reducing the fertility of females. Here, we study Wolbachia infection in a Drosophila parasitoid, Leptopilina heterotoma (Hymenoptera: Figitidae). A survey of Wolbachia infection shows that all natural populations tested are totally infected. Crosses between infected males and cured females show complete incompatibility: almost no females are produced. Moreover, incompatible eggs die early during their development, unlike Nasonia. This early death allows the parasitized Drosophila larva to achieve its development and to emerge. Thus, uninfected females crossed with infected males have reduced offspring production consisting only of males. Evidence of this CI type in insects demonstrates that the difference in CI types of Nasonia and Tetranychus is not due to specific factors of insects or acari. Using theoretical models, we compare the invasion processes of different strategies of Wolbachia: CI in diploid species, the two CI types in haplodiploid species, and parthenogenesis (the classical effect in haplodiploid species). Models show that CI in haplodiploid species is less efficient than in diploid ones. However, the Leptopilina type is advantageous compared to the Nasonia type. Parthenogenesis may be more or less advantageous, depending on the infection cost and on the proportion of fertilized eggs. Finally, we can propose different processes of Wolbachia strategy evolution in haplodiploid species from Nasonia CI type to Leptopilina CI type or parthenogenesis.

Physiological cost induced by the maternally-transmitted endosymbiont Wolbachia in the Drosophila parasitoid Leptopilina heterotoma

Endosymbiotic bacteria of the genus Wolbachia infect a number of invertebrate species in which they induce various alterations in host reproduction, mainly cytoplasmic incompatibility (CI). In contrast to most other maternally transmitted parasites, manipulation of host reproduction makes the spread of Wolbachia possible even if they induce a physiological cost on their hosts. Current studies have shown that fitness consequences of Wolbachia infection could range from positive (mutualist) to negative (parasitic) but, in most cases, Wolbachia do not have strong deleterious effects on host fitness and the status of association remains unclear. Here, we show that in the Drosophila parasitoid wasp Leptopilina heterotoma, Wolbachia infection has a negative impact on several host fitness traits of both sexes. Fecundity, adult survival and locomotor performance are significantly reduced, whereas circadian rhythm, development time and offspring sex-ratio are not affected. Although the cost of bacterial infection can be overcome by effects on host reproduction i.e. cytoplasmic incompatibility, it could influence the spread of the bacterium at the early stages of the invasion process. Clearly, results underline the wide spectrum of phenotypic effects of Wolbachia infection and, to our knowledge, Wolbachia infection of L. heterotoma appears to be one of the most virulent that has ever been observed in insects.

Ecological and genetic interactions in Drosophila-parasitoids communities: a case study with D. melanogaster, D. simulans and their common Leptopilina parasitoids in south-eastern France

Drosophila species are attacked by a number of parasitoid wasps, which constitute an important factor of population regulation. Since Drosophila melanogaster and Drosophila simulans share common parasitoid species, their ecology and evolution can hardly be understood without considering parasitoids. After a short review of data available on Drosophila–parasitoid interactions involving D. melanogaster and D. simulans as hosts, we report field and laboratory experiments investigating the ecological role of Leptopilina parasitoids in Drosophila communities of southern France. Seasonal survey of species abundance shows that strong interspecific interactions occur at both tropic levels. D. simulans progressively replaces D. melanogaster in southern areas suggesting competitive displacement. Parasitoids are responsible for very high Drosophila mortality (up to 90% in some fruits). Field data emphasize the importance of selective pressure that parasitoids exert on Drosophila communities. The two Leptopilina parasites (L. heterotoma and L. boulardi) have different local abundances, which vary in time, and they also compete for hosts. We show that parasitoids can mediate the coexistence of D. melanogaster and D. simulans in the laboratory, and thus may contribute to their puzzling coexistence in the field. Conversely, hosts exert selective pressures on parasitoids, and development on either D. melanogaster or D. simulans strongly affects fitness of adult wasps in a temperature-dependent fashion. Local variation in host species abundance and diversity could thus account for the genetic differentiation we observed in one parasitoid species. Despite laboratory studies cannot fully explain complex field situations, it is clear that the ecology and evolution of Drosophila populations and communities, especially D. melanogaster and D. simulans, are strongly constrained by parasitoids, which should receive more attention.

Ecology and life history evolution of frugivorous Drosophila parasitoids.

Parasitoids and their hosts are linked by intimate and harmful interactions that make them well suited to analyze fundamental ecological and evolutionary processes with regard to life histories evolution of parasitic association. Drosophila aspects of what parasitoid Hymenoptera have become model organisms to study aspects that cannot be investigated with other associations. These include the genetic bases of fitness traits variations, physiology and genetics of resistance/virulence, and coevolutionary dynamics leading to local adaptation. Recent research on evolutionary ecology of Drosophila parasitoids were performed mainly on species that thrive in fermenting fruits (genera Leptopilina and Asobara). Here, we review information and add original data regarding community ecology of these parasitoids, including species distribution, pattern of abundance and diversity, host range and the nature and intensity of species interactions. Biology and the evolution of life histories in response to habitat heterogeneity and possible local adaptations leading to specialization of these wasps are reported with special emphasis on species living in southern Europe. We expose the diversity and intensity of selective constraints acting on parasitoid life history traits, which vary geographically and highlight the importance of considering both biotic and abiotic factors with their interactions to understand ecological and evolutionary dynamics of host-parasitoid associations.

Adaptive significance of a circadian clock: temporal segregation of activities reduces intrinsic competitive inferiority in Drosophila parasitoids.

Most organisms show self–sustained circadian oscillations or biological clocks which control their daily fluctuations in behavioural and physiological activities. While extensive progress has been made in understanding the molecular mechanisms of biological clocks, there have been few clear demonstrations of the fitness value of endogenous rhythms. This study investigated the adaptive significance of circadian rhythms in a Drosophila parasitoid community. The activity rhythms of three sympatric Drosophila parasitoids are out of phase, the competitively inferior parasitoid species being active earlier than the superior competitor. This temporal segregation appears at least partially determined by endogenous periods of the clock which also vary between species and which correlate the time of activity. This earlier activity of the inferior competitor significantly reduces its intrinsic competitive disadvantage when multiparasitism occurs, thus suggesting that natural selection acting on the phase of the rhythm could substantially deviate the endogenous period from the optimal ca. 24 h period. This study demonstrates that temporal segregation of competing species could be endogenously controlled, which undoubtedly favours their coexistence in nature and also shows how natural selection can act on biological clocks to shape daily activity patterns.

Effects of temporary host deprivation on the reproductive potential of Trichogramma brassicae

The effect of host deprivation on the reproductive potential and parasitization behaviour of Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) was investigated. Upon emergence, females were deprived of hosts for one to four days, and were subsequently provided with an unlimited number of hosts. Regardless of its duration, deprivation did not affect female longevity, overall rate of progeny emergence, or progeny sex‐ratio. As deprivation time increased, however, the total number of parasitized hosts decreased from an average of 81.2 when hosts were provided at emergence, to an average of 34.0 when the waiting time was 4 days. This reduction was mainly due to the reduction in the parasitizations following the first laying peak, which was not affected when deprivation lasted fewer than 4 days. Trichogramma females are able to store ripe eggs (about 35) for 3 days without damage, but resorption occurs after four days of deprivation. Our results suggest that imaginal vitellogenesis occurs in this proovigenic wasp but that it is affected by host deprivation. The behaviour of females, as measured by the average distance between all parasitized hosts, was also modified by host deprivation. When deprivation exceeded one day, parasitizations were more scattered over the host patch. Moreover, increased deprivation times also appeared to delay the initiation of parasitization by a few hours.