Project description:Parasitoid wasps of the species Diachasmimorpha longicaudata are associated with a heritable poxvirus, known as DlEPV, that is stored in the venom gland of adult female wasps and transferred to tephritid fly hosts of the wasps during oviposition. We conducted a RNA-seq differential expression analysis to gain insight on how DlEPV can replicate in both wasps and their fly hosts but only cause pathogenic effects during replication in flies. Our analysis revealed that 91.2% (176 of 193) of DlEPV genes showed significant differential expression during peak virus replication in wasp venom glands compared to parasitized flies. Over 80% of DlEPV replication genes were significantly upregulated in wasps, while 79% of DlEPV putative virulence genes were significantly upregulated in fly hosts. These data therefore support a dichotomy of viral function, where virus replication is promoted in wasp tissue and virulence in host tissue. Such a division of viral activity could represent an important adaptation to maintain a stable symbiosis between this virus and its associated parasitoid.
Project description:Here we reveal evidence of a shared genetic toolkit across the full spectrum of social complexity found in Vespid wasps, from simple group living where castes remain plastic throughout life, to complex superorganismal societies comprised of mutually dependent insects with irreversible castes determined during development. We generated brain transcriptomic data for castes in nine representative species; using a machine learning approach we identified thousands of shared orthologs which consistently describe castes (queens and workers), from species with the simplest (Mischocyttarus paper wasps) to the most complex (e.g. Vespine wasps) levels of social organisation. The top 400 genes were enriched in synaptic transport genes, suggesting that these changes could affect brain neural function and connectivity. Fine-scale dissection of these patterns revealed that the molecular processes underpinning the simpler societies (which likely represent the origins of social living) are conserved throughout the major transition, but that additional processes may come into play in the more complex societies, especially at the point of no return where societies transition to be committed superorganisms. These analyses provide the first evidence of a conserved toolkit regulating social behaviour across the full spectrum of social complexity in any social insect. Importantly, they also provide evidence that there may be fundamental differences discriminating superorganismal societies from non-superorganismal societies. We suggest that the evolution of irreversible caste commitment (in superorganisms) is accompanied by a fundamental shift in the underlying regulatory molecular machinery; such shifts may also typify other major evolutionary transitions that are characterised by the emergence of a committed division of labour, such as the evolution of multicellularity.
Project description:No evidence of enemy release in pathogen and microbial communities of common wasps (Vespula vulgaris) in their native and introduced range
Project description:Although host-parasitoid interactions are becoming well characterized at the organismal and cellular levels, much remains to be understood of the molecular bases for the host immune response and the parasitoidsâ ability to defeat this immune response. Leptopilina boulardi and L. heterotoma, two closely related, highly infectious natural parasitoids of Drosophila melanogaster, appear to use very different infection strategies at the cellular level. Here, we further characterize cellular level differences in the infection characteristics of these two wasp species using newly derived, virulent inbred strains, and then use whole genome microarrays to compare the transcriptional response of Drosophila to each. While flies attacked by the melanogaster group specialist Leptopilina boulardi (strain Lb17) up-regulate numerous genes encoding proteolytic enzymes, components of the Toll and JAK/STAT pathways, and the melanization cascade as part of a combined cellular and humoral innate immune response, flies attacked by the generalist L. heterotoma (strain Lh14) do not appear to initiate an immune transcriptional response at the time points post-infection we assayed, perhaps due to the rapid venom-mediated lysis of host hemocytes (blood cells). Thus, the specialist parasitoid appears to invoke a full-blown immune response in the host, but suppresses and/or evades downstream components of this response. Given that activation of the host immune response likely depletes the energetic resources of the host, the specialistâs infection strategy seems relatively disadvantageous. However, we uncover the mechanism for one potentially important fitness tradeoff of the generalistâs highly immune suppressive infection strategy. Experiment Overall Design: The parasitoid wasps L. boulardi and L. heterotoma were allowed to attack late second instar D. melanogaster larvae (72 hrs old at 22ËC) in the following manner. Nine petri dishes containing 60 fly larvae were each exposed to six experienced L. boulardi (strain Lb17) female wasps for 2 hrs, another nine plates were exposed to five L. heterotoma (strain Lh14) females, and nine control plates were left uninfected. For each of three time points post-infection (2-5 hrs, 9-12 hrs, 21-24 hrs), 40 larvae from three replicate plates were removed and frozen at -80ËC for RNA extraction and microarray analysis (3 treatments x 3 time points x 3 replicates = 27 samples total).
Project description:One of the essential or beneficial micronutrient for plants and animals is boron that is an ultra-trace element. Although boron can inhibit the growth of Saccharomyces cerevisiae around 80 mM, it is also a growth supplement. However, little information is currently available regarding the molecular mechanisms and essentiality of boron. In this paper, the approach was to generate S. cerevisiae mutants with high boron resistance by using evolutionary engineering strategy that was previously applied successfully. Boron-resistant S. cerevisiae mutants were obtained and their phenotypic and physiological characteristics were determined. In order to identify the molecular mechanisms implicated in boron resistance, the whole transcriptomes and genome sequence analysis of wild type and one of the most resistant mutants were compared.
Project description:Goal: Natural populations of Drosophila melanogaster vary widely in their capacity to resist infection by the parasitic wasp Leptopilina boulardi. To date, little is known about the genetic, cellular and molecular basis underpinning the variation in susceptibility to parasitic wasps. In D.melanogaster populations artificially selected for parasitoid resistance, an increase in the number of circulating hemocytes (blood cells) is observed. One possibility is that this is accompanied by changes in hemocyte activation state, resulting in a successful defence response when exposed to parasitic wasps. We tested this possibility by generating populations that are highly resistant and highly susceptible to the parasitic wasps. We generated these populations after 26 generations of experimental evolution, starting from a single interbred wild caught population. Here, we study how selection for resistance to the parasitic wasp L. boulardi changes the number and type of circulating hemocytes in D. melanogaster larvae using scRNA-seq. Methods: Wild caught D. melanogaster females were collected from Cambridge, UK, to establish an outbreed population. From this, three replicated populations were selected for resistance to L. boulardi strain NSRef for 26 generations (Selection). Another three populations were maintained in the laboratory for 26 generations without being exposed to parasitoid-associated selection pressures (No Selection). At generation 26, second instar D. melanogaster larvae (48-63 hours after fertilization) from each population were infected with L. boulardi for three hours (Infection) or maintained without infection (No infection). 48 hours after, circulating hemocytes from third instar D. melanogaster larvae (96-111 hours after fertilization) from each population were collected in PBS and cleaned in OptiPrep solution (1.09g/ml). 10X Single Cell GEX v2 libraries were prepared and sequenced. CellRanger v2 was used to generate sample cell count matrices. Seurat v3 was used to integrate, normalise and cluster the cell types. Results: We identified novel plasmatocyte cell types, immature and mature lamellocytes and crystal cells. We identify a single lineage of cellular differentiation starting from a self-cycling plasmatocyte population, enriched for genes involved in extracellular matrix organisation, and ending in a mature lamellocyte population that has enriched expression of genes involved in signalling, hemostasis and gluconeogenesis. Immature lamellocytes were overrepresented in populations selected for resistance to parasitoids. Infection with parasitoid resulted in an increase in both immature and mature lamellocytes in circulation. Conclusions: Selection for resistance to the parasitic wasps resulted in the constitutive activation of the D. melanogaster immune system, where plasmatocytes differentiate into immature lamellocytes even where they are not exposed to parasitic wasps. We speculate these constitutive changes assist in mounting a successful defence response when exposed to parasitic wasps.
Project description:Oak galls form when gall wasps lay their eggs into part of the tree; in some galls, this attachment point to the host consists of only a few cells. The gall itself comprises entirely of host tissue; however, the initiation, development, and physical appearance are controlled by the inducer. This raises the intriguing question of the molecular mechanisms underlying gall formation, by which one or a small number of cells are reprogrammed and commit to a novel developmental path. Gall wasps undergo two generations each year, and the galls formed by these two generations exhibit markedly different appearances. We sequenced the transcriptomes of both the sexual and asexual generations of Neuropterus quercusbaccarum and Neuroterus numismalis. The transcriptomes of the generations that occur at the same time of year are more similar to each other than they are to the opposite generation of their respective species.
Project description:Trained immunity is the heightened state of innate immune memory that enhances immune response resulting in nonspecific protection. Epigenetic changes and metabolic reprogramming are critical steps that regulate trained immunity. In this study, we reported the involvement of O6-methylguanine DNA methyltransferase (MGMT), a DNA repair enzyme of lesion induced by alkylating agents, in regulation the trained immunity induced by β-glucan (BG). Pharmacological inhibition or silencing of MGMT expression altered LPS stimulated pro-inflammatory cytokine productions in BG-trained bone marrow derived macrophages (BMMs). Targeted deletion of Mgmt in BMMs resulted in reduction of the trained responses both in vitro and in vivo models. The transcriptomic analysis revealed that the dampening trained immunity in MGMT KO BMMs is partially mediated by ATM/FXR/AMPK axis affecting the MAPK/mTOR/HIF1α pathways and the reduction in glycolysis function. Taken together, a failure to resolve a DNA damage may have consequences for innate immune memory.