Wasp Infestation Research Articles

Inhibition of the Notch signal transducer CSL by Pkc53E-mediated phosphorylation to fend off parasitic immune challenge in Drosophila.

Notch signalling activity regulates hematopoiesis in Drosophila and vertebrates alike. Parasitoid wasp infestation of Drosophila larvae, however, requires a timely downregulation of Notch activity to allow the formation of encapsulation-active blood cells. Here, we show that the Drosophila CSL transcription factor Suppressor of Hairless [Su(H)] is phosphorylated at Serine 269 in response to parasitoid wasp infestation. As this phosphorylation interferes with the DNA binding of Su(H), it reversibly precludes its activity. Accordingly, phospho-deficient Su(H)S269A mutants are immune-compromised. A screen for kinases involved in Su(H) phosphorylation identified Pkc53E, required for normal hematopoiesis as well as for parasitoid immune response. Genetic and molecular interactions support the specificity of the Su(H)-Pkc53E relationship. Moreover, phorbol ester treatment inhibits Su(H) activity in vivo and in human cell culture. We conclude that Pkc53E targets Su(H) during parasitic wasp infestation, thereby remodelling the blood cell population required for wasp egg encapsulation.

Ecological Impact of American Chestnut Hybrid Restoration on Invertebrate Communities Above- and Belowground

The cross-hybridization of American chestnut (Castanea dentata (Marsh.) Borkh.) with Chinese chestnut (Castanea mollissima Bl.) is a promising strategy for restoring a blight-resistant strain of this keystone species to the Appalachian mountains. To assess the ecological impacts of hybridization on invertebrate communities, we conducted a study across chestnut plots with varying degrees of hybridization (75%, 94%, or 100% American chestnut). Our findings indicate American chestnut hybridization impacted invertebrate communities above- and belowground. Aboveground insect community composition, insect herbivory, gall infestation, and belowground invertebrate diversity were all altered. While some of these differences could be explained by different growth habits or environmental differences, stark differences in Asian chestnut gall wasp infestation (Dryocosmus kuriphilus Yasumatsu.) suggest a genetic component. These results suggest that chestnut hybridization, and particularly expanded restoration efforts using chestnut hybrids, could impact invertebrate communities above- and belowground in addition to pest dynamics. Understanding these effects is crucial for successful chestnut restoration and ecosystem management.

The Endosomal Sorting Complex, ESCRT, has diverse roles in blood progenitor maintenance, lineage choice and immune response.

Most hematological malignancies are associated with reduced expression of one or more components of the Endosomal Sorting Complex Required for Transport (ESCRT). However, the roles of ESCRT in stem cell and progenitor maintenance are not resolved. Parsing signaling pathways in relation to the canonical role of ESCRT poses a challenge. The Drosophila hematopoietic organ, the larval lymph gland, provides a path to dissect the roles of cellular trafficking pathways such as ESCRT in blood development and maintenance. Drosophila has 13 core ESCRT components. Knockdown of individual ESCRTs showed that only Vps28 and Vp36 were required in all lymph gland progenitors. Using the well-conserved ESCRT-II complex as an example of the range of phenotypes seen upon ESCRT depletion, we show that ESCRTs have cell-autonomous as well as non-autonomous roles in progenitor maintenance and differentiation. ESCRT depletion also sensitized posterior lobe progenitors to respond to immunogenic wasp infestation. We also identify key heterotypic roles for ESCRT in position-dependent control of Notch activation to suppress crystal cell differentiation. Our study shows that the cargo sorting machinery determines the identity of progenitors and their adaptability to the dynamic microenvironment. These mechanisms for control of cell fate may tailor developmental diversity in multiple contexts.

JAK/STAT mediated insulin resistance in muscles is essential for effective immune response

BackgroundThe metabolically demanding nature of immune response requires nutrients to be preferentially directed towards the immune system at the expense of peripheral tissues. We study the mechanisms by which this metabolic reprograming occurs using the parasitoid infection of Drosophila larvae. To overcome such an immune challenge hemocytes differentiate into lamellocytes, which encapsulate and melanize the parasitoid egg. Hemocytes acquire the energy for this process by expressing JAK/STAT ligands upd2 and upd3, which activates JAK/STAT signaling in muscles and redirects carbohydrates away from muscles in favor of immune cells.MethodsImmune response of Drosophila larvae was induced by parasitoid wasp infestation. Carbohydrate levels, larval locomotion and gene expression of key proteins were compared between control and infected animals. Efficacy of lamellocyte production and resistance to wasp infection was observed for RNAi and mutant animals.ResultsAbsence of upd/JAK/STAT signaling leads to an impaired immune response and increased mortality. We demonstrate how JAK/STAT signaling in muscles leads to suppression of insulin signaling through activation of ImpL2, the inhibitor of Drosophila insulin like peptides.ConclusionsOur findings reveal cross-talk between immune cells and muscles mediates a metabolic shift, redirecting carbohydrates towards immune cells. We emphasize the crucial function of muscles during immune response and show the benefits of insulin resistance as an adaptive mechanism that is necessary for survival.

Single-cell transcriptomics identifies new blood cell populations in Drosophila released at the onset of metamorphosis.

Drosophila blood cells called hemocytes form an efficient barrier against infections and tissue damage. During metamorphosis, hemocytes undergo tremendous changes in their shape and behavior, preparing them for tissue clearance. Yet, the diversity and functional plasticity of pupal blood cells have not been explored. Here, we combine single-cell transcriptomics and high-resolution microscopy to dissect the heterogeneity and plasticity of pupal hemocytes. We identified undifferentiated and specified hemocytes with different molecular signatures associated with distinct functions such as antimicrobial, antifungal immune defense, cell adhesion or secretion. Strikingly, we identified a highly migratory and immune-responsive pupal cell population expressing typical markers of the posterior signaling center (PSC), which is known to be an important niche in the larval lymph gland. PSC-like cells become restricted to the abdominal segments and are morphologically very distinct from typical Hemolectin (Hml)-positive plasmatocytes. G-TRACE lineage experiments further suggest that PSC-like cells can transdifferentiate to lamellocytes triggered by parasitoid wasp infestation. In summary, we present the first molecular description of pupal Drosophila blood cells, providing insights into blood cell functional diversification and plasticity during pupal metamorphosis.

Wild olive seed weevil, Anchonocranus oleae Marshall (Coleoptera: Curculionidae), in cultivated olives in South Africa

Wild olive seed weevil, Anchonocranus oleae Marshall, larvae as well as oviposition and feeding damage were recorded in cultivated olives during a recent study on olive seed wasps. Prematurely dropped fruit and fruit picked at harvest in two orchards near Stellenbosch and Agter-Paarl were examined regularly for olive seed wasp infestation over three seasons. In addition, olives were collected in the olive growing regions of the Western Cape province for a survey to determine olive seed wasp distribution. DNA barcoding was used to confirm the identity of weevil larvae in kernels. The number of A. oleae larvae found in olive seeds and the number of olives with weevil oviposition or feeding damage in this study were low. The presence of larvae and weevil damage in two orchards in the survey with no wild olive trees in close proximity suggest that the weevil could breed and persist in cultivated olive orchards. Currently A. oleae is not of economic concern, but if infested olives are discarded at harvest and left in orchards, the weevils could complete their development in the kernels and numbers could eventually increase to damaging levels.

Leaf Trait Hyperspectral Characterization of Castanea sativa Miller Affected by Dryocosmus kuriphilus Yasumatsu

While populations of the Asian chestnut gall wasp (Dryocosmus kuriphilus Yasumatsu), an invasive pest affecting the European chestnut (Castanea sativa Miller), have started to be controlled biologically, this pest still conditions chestnut tree development. With the aim of assessing plant health status as a means of monitoring gall wasp infestation, we used a field spectroradiometer to collect data from leaves taken from 83 trees in two chestnut orchards. We calculated characteristic spectral signatures for pest infestation, and after training and validation, developed classifiers to distinguish between different infestation levels. Several partial least square discriminant analysis (PLS-DA) and random forest (RF) models were fitted with reflectance and transformed values to obtain characteristic curves reflecting infestation. Four wavelengths (560 nm, 680 nm, 1400 nm, and 1935 nm) were identified as showing the greatest differences between curves. The best overall accuracy (69.23%) was achieved by an RF model fitted with reflectance transformed values. Lower overall accuracy (26.92%) was achieved in distinguishing between infestation levels. In conclusion, while more specific differences in infestation levels were not detectable, our method successfully discriminated between gall absence and presence.

Transcriptome Analysis Reveals Genes Involved in Responses of Eucalyptus to Gall Wasp Infestation

Leptocybe invasa is a gall wasp causing significant damage to Eucalyptus species. This study used RNA sequencing (RNA-seq) to identify differentially expressed genes (DEGs) associated with early L. invasa infestation in eucalyptus parents and their F1-progenies. A total of 14,648 significant DEGs were identified from U22-tolerant and C153-susceptible parents, and extremely tolerant and susceptible pools of their F1-progenies. A total of 324/632 and 182/205 DEGs specific for the tolerant group and the susceptible group were up-regulated, respectively. Expression analysis by qRT-PCR of the selected DEGs was comparable with the results of RNA-seq. Expression analysis of the selected genes using the top five progenies from each pool was consistent with that in the parents. Three genes (RCA, SUI1, GCN5) were up-regulated after infestation in all tested tolerant plants, suggesting their important roles in the tolerant phenotype. Using expression and STRING analysis, our results suggest that early response at three days after gall wasp infestation increased protein and terpenoid synthesis, and increased transportation of these molecules. In addition, wounding also increased photosynthesis and glycolysis. These processes involved the interaction of several plant hormones, such as JA, Auxin, and ABA. The information obtained from this study could be useful for future Eucalyptus breeding programs.

The impact of the Asian chestnut gall wasp (Dryocosmus kuriphilus) on chestnut tree growth may be mediated by site resources

IntroductionThe Asian chestnut gall wasp (Dryocosmus kuriphilus) is a major pest of chestnut trees worldwide, seriously affecting chestnut cultivation. Information concerning the effects of gall wasp attack on diameter growth of chestnut trees is currently scarce and limited to coppice stands and to use of the growth of a non-target control species (unaffected by the pest) for reference purposes. The effects of the pest on widely-spaced plantations (grown at a much lower density than chestnut coppices) and the use of explicitly-observed annual infestation rate data remain to be explored.MethodsIn the present study, we analyzed the impact of the chestnut gall wasp on the diameter growth of chestnut trees, using data from 16 experimental plots established in widely-spaced plantations located in good quality sites. Two of the plots are in plantations where a susceptible hybrid chestnut clone and a chestnut clone resistant to the gall wasp coexist, whereas the remaining 14 plots are in Castanea sativa plantations where the level of gall wasp infestation varies across trees and years. The plots were surveyed to determine the diameter growth of the trees and the level of infestation during 5 years (2017–2021).ResultsThe infestation level corresponding to the theoretical damage threshold was surpassed inmost plots during the study period. Nevertheless, there were no differences in the growth of attacked and unaffected plants in the two plots planted with hybrid clones with contrasting susceptibility to the gall wasp. The attack had a modest effect in C. sativa plots, with a mean reduction in annual basal area increment of 9.9%.DiscussionThese findings apparently contradict previous reports of a marked reduction in radial growth of chestnut coppice trees due to gall wasp attack. The difference in findings may be related to increased compensation for herbivory with increasing levels of resources (especially light) in the plantations under study, which were less dense than previously studied stands. The study outcomes add to existing knowledge on the impact of chestnut gall wasp on wood formation and may have implications regarding planting site recommendations and subsequent stand management.

Methotrexate negatively acts on inflammatory responses triggered in Drosophila larva with hyperactive JAK/STAT pathway

Drosophila is a valuable paradigm for studying tumorigenesis and cancer. Mutations causing hematopoietic aberrations and melanotic-blood-tumors found in Drosophila mutants are vastly studied. Clear understanding about the blood cells, signaling pathways and the tissues affected during hematopoietic tumor formation provide an opportunity to delineate the effects of cancer therapeutics. Using this simple hematopoietic archetype, we elucidated the effects of the anti-cancer drug, Methotrexate (MTX) on immune responses in two scenarios i.e. against wasp infection and in hematopoietic mutant, hopTum-l. Through this in vivo study we show that MTX impedes the immune responses against wasp infection including the encapsulation response. We further observed that MTX reduces the tumor penetrance in gain-of-function mutants of JAK/STAT pathway, hopTum-l. MTX is anti-inflammatory as it hinders not only the immune responses of acute inflammation as observed after wasp infestation, but also chronic inflammatory responses associated with constitutively activated JAK/STAT pathway mutant (hopTum-l) carrying blood tumors.

Single-cell transcriptome maps of myeloid blood cell lineages in Drosophila

The Drosophila lymph gland, the larval hematopoietic organ comprised of prohemocytes and mature hemocytes, has been a valuable model for understanding mechanisms underlying hematopoiesis and immunity. Three types of mature hemocytes have been characterized in the lymph gland: plasmatocytes, lamellocytes, and crystal cells, which are analogous to vertebrate myeloid cells, yet molecular underpinnings of the lymph gland hemocytes have been less investigated. Here, we use single-cell RNA sequencing to comprehensively analyze heterogeneity of developing hemocytes in the lymph gland, and discover previously undescribed hemocyte types including adipohemocytes, stem-like prohemocytes, and intermediate prohemocytes. Additionally, we identify the developmental trajectory of hemocytes during normal development as well as the emergence of the lamellocyte lineage following active cellular immunity caused by wasp infestation. Finally, we establish similarities and differences between embryonically derived- and larval lymph gland hemocytes. Altogether, our study provides detailed insights into the hemocyte development and cellular immune responses at single-cell resolution.

Biological control of eucalyptus Gall wasp, Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae) in Punjab, India

In this investigation, biological control of eucalyptus Gall wasp, Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae) in Punjab, India was investigated. The genus Leptocybe Fisher & La Salle (Hymenoptera: Eulophidae) was founded with its type species, L. invasa from Israel in 2004. Adult wasps of L. invasa induce galls on the shoots and midribs of leaves of Eucalyptus sp. and also damage nursery seedlings and young plantations. In the present study, survey was carried out from the year 2012 to 2014, on gall wasp infestation in different eucalyptus growing areas of Punjab. Observations revealed that eucalyptus growing areas in Punjab were under the threat of attack by gall wasp, L. invasa. During the present study, biological control agents Megastimus viggianii and Quadrastichus mendeli have been successfully applied in the gall infested areas of Punjab, from 2012 to 2017, where infestation of gall wasp was regularly decreased and came below 10 % by July, 2017. Consequently, Eucalyptus gall forming insect L. invasa has been controlled successfully by applying the biological control through releasing the parasitoids (Megastimus viggianii and Quadrastichus mendeli), in Punjab, India.

Temporal specificity and heterogeneity of Drosophila immune cells.

Immune cells provide defense against non-self and have recently been shown to also play key roles in diverse processes such as development, metabolism, and tumor progression. The heterogeneity of Drosophila immune cells (hemocytes) remains an open question. Using bulk RNA sequencing, we find that the hemocytes display distinct features in the embryo, a closed and rapidly developing system, compared to the larva, which is exposed to environmental and metabolic challenges. Through single-cell RNA sequencing, we identify fourteen hemocyte clusters present in unchallenged larvae and associated with distinct processes, e.g., proliferation, phagocytosis, metabolic homeostasis, and humoral response. Finally, we characterize the changes occurring in the hemocyte clusters upon wasp infestation, which triggers the differentiation of a novel hemocyte type, the lamellocyte. This first molecular atlas of hemocytes provides insights and paves the way to study the biology of the Drosophila immune cells in physiological and pathological conditions.

Subpopulation of Macrophage-Like Plasmatocytes Attenuates Systemic Growth via JAK/STAT in the Drosophila Fat Body.

Drosophila hemocytes, like those of mammals, are given rise from two distinctive phases during both the embryonic and larval hematopoiesis. Embryonically derived hemocytes, mostly composed of macrophage-like plasmatocytes, are largely identified by genetic markers. However, the cellular diversity and distinct functions of possible subpopulations within plasmatocytes have not been explored in Drosophila larvae. Here, we show that larval plasmatocytes exhibit differential expressions of Hemolectin (Hml) and Peroxidasin (Pxn) during development. Moreover, removal of plasmatocytes by overexpressing pro-apoptotic genes, hid and reaper in Hml-positive plasmatocytes, feeding high sucrose diet, or wasp infestation results in increased circulating hemocytes that are Hml-negative. Interestingly these Hml-negative plasmatocytes retain Pxn expression, and animals expressing Hml-negative and Pxn-positive subtype largely attenuate growth and abrogate metabolism. Furthermore, elevated levels of a cytokine, unpaired 3, are detected when Hml-positive hemocytes are ablated, which in turn activates JAK/STAT activity in several tissues including the fat body. Finally, we observed that insulin signaling is inhibited in this background, which can be recovered by concurrent loss of upd3. Overall, this study highlights heterogeneity in Drosophila plasmatocytes and a functional plasticity of each subtype, which reaffirms extension of their role beyond immunity into metabolic regulation for cooperatively maintaining internal homeostatic balance.

Habitat factors associated with Fopius caudatus parasitism and population level of its host, Ceratitis cosyra

AbstractBiotic and abiotic factors affect herbivores and their natural enemies and understanding of their requirements may permit habitat modification enabling conservation biological control. Ceratitis cosyra Walker (Diptera: Tephritidae), an African‐native fruit fly pest is mostly parasitized by the parasitoid wasp Fopius caudatus Szépligeti (Hymenoptera: Braconidae). To assess F. caudatus habitat, the wasp parasitism levels and infestation of its fruit fly host were examined in Sarcocephalus latifolius (Smith) Bruce (Rubiaceae), a shrub of which the fruits are among the preferred hosts of C. cosyra and F. caudatus. Fruit‐collection site descriptions, including plant species presence, were analysed in relation to the target insect abundances (emergence from target fruit). Ceratitis cosyra and F. caudatus emerged from all sites; nonetheless, their population levels were associated with both abiotic and biotic factors, of which some can be manipulated. Several factors, such as cultivation level, topography, and vegetation coverage, were correlated with F. caudatus parasitism. Ceratitis cosyra infestation level was correlated with factors such as density of S. latifolius, vegetation cover, cultivation practices, temperature, altitude, rainfall pattern, and stoniness. Proximity to other fruit fly host plants correlated with both pest abundance and F. caudatus parasitism level of the fruit fly. The findings that insects’ interactions and abundance are influenced by habitat structure and that parasitism is positively related to natural habitat indicates the importance of maintaining natural habitats in closeness to cultivated areas with the aim of enhancing pest suppression by parasitoids. Further studies should attempt to identify how plant species composition in and around orchards could affect the management of tephritid fruit fly pests.

Torymus sinensis Kamijo, a biocontrol agent against the invasive chestnut gall wasp Dryocosmus kuriphilus Yasumatsu in Spain: its natural dispersal from France and the first data on establishment after experimental releases

Aim of study: The globally invasive gall wasp, Dryocosmus kuriphilus Yasumatsu, 1951 (Cynipidae: Cynipini), reached Spain seven years ago and is already regarded as an important pest of chestnuts (Castanea spp.) in this country as well as worldwide. In this paper, we present comprehensive data on the establishment in Spain of Torymus sinensis Kamijo, 1982 (Chalcidoidea: Torymidae), an effective non-native natural enemy of this pest, as a result of both natural dispersal and settlement after controlled releases since 2015.Area of study: Sites of the Spanish autonomous communities of Galicia, Asturias, Basque Country, Catalonia, Andalusia and Madrid where D. kuriphilus is present.Material and methods: To study the natural dispersal of T. sinensis from France, we selected two sampling sites in Catalonia, six in the Basque Country and two in Navarra known for their heavy Asian chestnut gall wasp (ACGW) infestation; to study T. sinensis establishment after authorized controlled releases by the concerned authorities, the field samplings were done mainly in Galicia (35 sites) and Andalusia (8 sites). Additionally an experimental release study was made in Madrid.Main Results: Our results showed that T. sinensis has spread throughout Spain by natural dispersal across the French border and now occurs in Catalonia (two sites), the Basque Country (three sites) and Navarra (one site) but not in the neighbouring region of Cantabria. The percentage of parasitism by T. sinensis on D. kuriphilus is higher in the Basque Country sites, which are close to the French border, thus indicating that its establishment in these localities is not recent. After controlled releases, T. sinensis has been successfully established in five release sites in Andalusia (Valle del Genal and Sierra Blanca, Málaga Province) and one release site in Madrid. However, in the region of Galicia (NW Spain), where the number of authorized releases has been higher, the establishment of T. sinensis still appears to be very low.Research highlights: Established populations of T. sinensis may exert a positive buffer against D. kuriphilus-driven chestnut infestation in Spain, similar to what is observed in other invaded European countries.Additional Keywords: Controlled releases, Torymidae, invasive species, Cynipidae, natural spread, biological control.Abbreviations used: Asian chestnut gall wasp (ACGW); Torymus sinensis parasitism rate (TsPR); mean of ACGW larval chambers per gall (MACGW).

Infestation potential of Dryocosmus kuriphilus Yasumatsu, 1951 (Hymenoptera: Cynipidae) in different natural populations of Castanea sativa Miller: an experimental ex situ test

The Asian chestnut gall wasp was accidentally introduced in Italy in 2002 and spread across Europe in the following years, becoming a serious threat for chestnut cultivations and rural economies of many countries. Exploring the variation in susceptibility of the host genetic resources is crucial to face the spreading of this pest. We used an experimental approach for testing the differential susceptibility within and between populations of European chestnut. For doing this, we compared both the infestation level and the rate of immune individuals in trees from populations of Spain, Italy, and Greece. We found that the level of infestation is not significantly different in the different provenances but that a higher rate of immune trees occur in Greece. Our results suggest that two different contingents of trees compose Greek populations: one major group of trees with the same susceptibility as the other populations and a second minor group of trees resistant to gall wasp infestation. Our data lay the basis for improving the currently adopted measures to mitigate gall wasp impacts.

Embryonic hematopoiesis modulates the inflammatory response and larval hematopoiesis in Drosophila.

Recent lineage tracing analyses have significantly improved our understanding of immune system development and highlighted the importance of the different hematopoietic waves. The current challenge is to understand whether these waves interact and whether this affects the function of the immune system. Here we report a molecular pathway regulating the immune response and involving the communication between embryonic and larval hematopoietic waves in Drosophila. Down-regulating the transcription factor Gcm specific to embryonic hematopoiesis enhances the larval phenotypes induced by over-expressing the pro-inflammatory Jak/Stat pathway or by wasp infestation. Gcm works by modulating the transduction of the Upd cytokines to the site of larval hematopoiesis and hence the response to chronic (Jak/Stat over-expression) and acute (wasp infestation) immune challenges. Thus, homeostatic interactions control the function of the immune system in physiology and pathology. Our data also indicate that a transiently expressed developmental pathway has a long-lasting effect on the immune response.

The nociception genes painless and Piezo are required for the cellular immune response of Drosophila larvae to wasp parasitization

In vertebrates, interaction between the nervous system and immune system is important to protect a challenged host from stress inputs from external sources. In this study, we demonstrate that sensory neurons are involved in the cellular immune response elicited by wasp infestation of Drosophila larvae. Multidendritic class IV neurons sense contacts from external stimuli and induce avoidance behaviors for host defense. Our findings show that inactivation of these sensory neurons impairs the cellular response against wasp parasitization. We also demonstrate that the nociception genes encoding the mechanosensory receptors Painless and Piezo, both expressed in class IV neurons, are essential for the normal cellular immune response to parasite challenge.

Screening and Analysis of Janelia FlyLight Project Enhancer-Gal4 Strains Identifies Multiple Gene Enhancers Active During Hematopoiesis in Normal and Wasp-ChallengedDrosophilaLarvae

A GFP expression screen has been conducted on >1000 Janelia FlyLight Project enhancer-Gal4 lines to identify transcriptional enhancers active in the larval hematopoietic system. A total of 190 enhancers associated with 87 distinct genes showed activity in cells of the third instar larval lymph gland and hemolymph. That is, gene enhancers were active in cells of the lymph gland posterior signaling center (PSC), medullary zone (MZ), and/or cortical zone (CZ), while certain of the transcriptional control regions were active in circulating hemocytes. Phenotypic analyses were undertaken on 81 of these hematopoietic-expressed genes, with nine genes characterized in detail as to gain- and loss-of-function phenotypes in larval hematopoietic tissues and blood cells. These studies demonstrated the functional requirement of the cut gene for proper PSC niche formation, the hairy, Btk29A, and E2F1 genes for blood cell progenitor production in the MZ domain, and the longitudinals lacking, dFOXO, kayak, cap-n-collar, and delilah genes for lamellocyte induction and/or differentiation in response to parasitic wasp challenge and infestation of larvae. Together, these findings contribute substantial information to our knowledge of genes expressed during the larval stage of Drosophila hematopoiesis and newly identify multiple genes required for this developmental process.