AANAT1 regulates insect midgut detoxification through the ROS/CncC pathway.

Insecticide resistance has been a problem in both the agricultural pests and vectors. Revealing the detoxification mechanisms may help to better manage insect pests. Here, we showed that arylalkylamine N-acetyltransferase 1 (AANAT1) regulates intestinal detoxification process through modulation of reactive oxygen species (ROS)-activated transcription factors cap"n"collar isoform-C (CncC): muscle aponeurosis fibromatosis (Maf) pathway in both the oriental fruit fly, Bactrocera dorsalis, and the arbovirus vector, Aedes aegypti. Knockout/knockdown of AANAT1 led to accumulation of biogenic amines, which induced a decreased in the gut ROS level. The reduced midgut ROS levels resulted in decreased expression of CncC and Maf, leading to lower expression level of detoxification genes. AANAT1 knockout/knockdown insects were more susceptible to insecticide treatments. Our study reveals that normal functionality of AANAT1 is important for the regulation of gut detoxification pathways, providing insights into the mechanism underlying the gut defense against xenobiotics in metazoans.

Characterization of an agmatine N-acetyltransferase from Bactrocera dorsalis that modulates ovary development.

Agmatine N-acetyltransferase (AgmNAT), which catalyzes the formation of N-acetylagmatine from acetyl-CoA and agmatine, is a member of the GCN5-related N-acetyltransferase family. So far, knowledge of the physiological roles of AgmNAT in insects is limited. Here, we identified one gene encoding protein homologous to that of Drosophila AgmNAT using sequence information from an activity-verified Drosophila AgmNAT in a BLAST search of the Bactrocera dorsalis genome. We expressed and purified B. dorsalis AgmNAT in Escherichia coli and used the purified enzyme to define the substrate specificity for acyl-CoA and amine substrates. Our application of the screening strategy to BdorAgmNAT led to the identification of agmatine as the best amine substrate for this enzyme, with the highest kcat/Km value. We successfully obtained a BdorAgmNAT knockout strain based on a wild-type strain (WT) using the CRISPR/Cas9 technique. The ovary development of the BdorAgmNAT knockout mutants was delayed for 10 days compared with the WT specimens. Moreover, mutants had a much smaller mature ovary size and laid far fewer eggs than WT. Loss of function of BdorAgmNAT caused by RNAi with mature WT females did not affect their fecundity. These findings indicate that BdorAgmNAT is critical for oogenesis. Our data provide the first evidence for AgmNAT in regulating ovary development.

Toxicity of Beauveria bassiana to Bactrocera dorsalis and effects on its natural predators.

Entomopathogenic fungi (EPF) are economical and environmentally friendly, forming an essential part of integrated pest management strategies. We screened six strains of Beauveria bassiana (B1-B6) (Hypocreales: Cordycipitaceae), of which B4 was the most virulent to Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). We further assessed the biological characteristics of strain B4 and the environmental factors influencing its ability to infect B. dorsalis. We also evaluated the effects of B4 on two of the natural predators of B. dorsalis. We found that strain B4 was the most virulent to 3rd instar larvae, pupae, and adult B. dorsalis, causing mortality rates of 52.67, 61.33, and 90.67%, respectively. B4 was not toxic to B. dorsalis eggs. The optimum B4 effects on B. dorsalis were achieved at a relative humidity of 91-100% and a temperature of 25°C. Among the six insecticides commonly used for B. dorsalis control, 1.8% abamectin emulsifiable concentrate had the strongest inhibitory effect on B4 strain germination. B4 spraying affected both natural enemies (Amblyseius cucumeris and Anastatus japonicus), reducing the number of A. cucumeris and killing A. japonicus adults. We found a valuable strain of EPF (B4) that is virulent against many life stages of B. dorsalis and has great potential for the biological control of B. dorsalis. We also provide an important theoretical and practical base for developing a potential fungicide to control B. dorsalis.

Arylalkylamine N-acetyltransferase 1 gene (AANAT1) regulates cuticle pigmentation and ovary development of the adult oriental fruit fly, Bactrocera dorsalis.

The arylalkylamine N-acetyltransferase (AANAT) enzymes catalyze the acetyl-CoA-dependent acetylation of an amine or arylalkylamine, which is involved in important biological processes of insects. Here, we carried out the molecular and biochemical identification of an arylalkylamine N-acetyltransferase (AANAT) from the oriental fruit fly, Bactrocera dorsalis. Using a bacterial expression system, we expressed and purified the encoded recombinant BdorAANAT1-V3 protein. The purified recombinant protein acts on a wide range of substrates, including dopamine, tyramine, octopamine, serotonin, methoxytryptamine, and tryptamine, and shows similar substrate affinity (i.e., Km values: 0.16-0.26 mM) except for serotonin (Km = 0.74 mM) and dopamine (Km = 0.84 mM). Transcriptional profile analysis of BdorAANAT1 revealed that this gene is most prevalent in adults and abundant in the adult brain, gut, and ovary. Using the CRISPR/Cas9 technique, we successfully obtained a BdorAANAT1 knockout strain based on a wild-type strain (WT). Compared with the WT, the cuticle color of larvae and pupae is normal; however, in adult mutants, the yellow region of their thorax is darkly pigmented, and two black spots were evident at the abdomen's end. Moreover, the female BdorAANAT1 knockout mutant had a smaller ovary than the WT, and laid far fewer eggs. Loss of function of BdorAANAT1 caused by RNAi with mature adult females in which the reproductive system is fully developed had no effect on their fecundity. Altogether, these results indicate that BdorAANAT1 regulates ovary development. Our findings provide evidence for the insect AANAT1 modulating adult cuticle pigmentation and female fecundity.

Serotonin modulates insect gut bacterial community homeostasis.

BACKGROUND: Metazoan guts are in permanent contact with microbial communities. However, the host mechanisms that have developed to manage the dynamic changes of these microorganisms and maintain homeostasis remain largely unknown. RESULTS: Serotonin (5-hydroxytryptamine [5-HT]) was found to modulate gut microbiome homeostasis via regulation of a dual oxidase (Duox) gene expression in both Bactrocera dorsalis and Aedes aegypti. The knockdown of the peripheral 5-HT biosynthetic gene phenylalanine hydroxylase (TPH) increased the expression of Duox and the activity of reactive oxygen species, leading to a decrease in the gut microbiome load. Moreover, the TPH knockdown reduced the relative abundance of the bacterial genera Serratia and Providencia, including the opportunistic pathogens, S. marcescens and P. alcalifaciens in B. dorsalis. Treatment with 5-hydroxytryptophan, a precursor of 5-HT synthesis, fully rescued the TPH knockdown-induced phenotype. CONCLUSIONS: The findings reveal the important contribution of 5-HT in regulating gut homeostasis, providing new insights into gut-microbe interactions in metazoans.

Identification of putative muscarinic acetylcholine receptor genes in Bactrocera dorsalis and functional analysis of Bdor-mAChR-B.

Muscarinic acetylcholine receptors (mAChRs) play important roles in the insect nervous system. These receptors are G protein-coupled receptors, which are potential targets for insecticide development. While the investigation of pharmacological properties of insect mAChRs is growing, the physiological roles of the receptor subtype remain largely indeterminate. Here, we identified three mAChR genes in an important agricultural pest Bactrocera dorsalis. Phylogenetic analysis defined these genes as mAChR-A, -B, and -C. Transcripts of the three mAChRs are most prevalent in 1-d-old larvae and are more abundant in the brain than other body parts in adults. Functional assay of Bdor-mAChR-B transiently expressed in Chinese hamster ovary cells showed that it was activated by acetylcholine (EC50, 205.11 nM) and the mAChR agonist oxotremorine M (EC50, 2.39 µM) in a dose-dependent manner. Using the CRISPR/Cas9 technique, we successfully obtained a Bdor-mAChR-B knockout strain based on wild-type (WT) strain. When compared with WT, the hatching and eclosion rate of Bdor-mAChR-B mutants are significantly lower. Moreover, the crawl speed of Bdor-mAChR-B knockout larvae was lower than that of WT, while climbing performance was enhanced in the mutant adults. Adults with loss of function of Bdor-mAChR-B showed declined copulation rates and egg numbers (by mated females). Our results indicate that Bdor-mAChR-B plays a key role in the development, locomotion, and mating behavior of B. dorsalis.

Intestinal responses of the oriental fruit fly Bactrocera dorsalis (Hendel) after ingestion of an entomopathogenic bacterium strain.

BACKGROUND: Bactrocera dorsalis (Hendel) is the main fruit fly pest of tropical and subtropical countries. The application of insecticides to manage this pest has led to serious resistance problems; therefore, new ways to control B. dorsalis are required. Pathogenic bacteria are sources of biocontrol agents for pest management. RESULTS: We determined that a pathogenic bacterial strain, Serratia marcescens PS-1, isolated from a moribund striped flea beetle (Phyllotreta striolata), was lethal to B. dorsalis adults following ingestion. Histological analyses revealed that PS-1 damaged the intestinal epithelium, resulting in cell death within 24 h. We then generated a gut transcriptomic data set using RNA-Seq at two time points (6 and 24 h) after PS-1 infection. We found that genes encoding the peritrophic matrix constituent were down-regulated, whereas genes involved in lipid and glycan metabolism, and renewal of the gut epithelium, along with genes encoding digestive enzymes and stress response factors, were up-regulated. In addition, 14 cecropin genes were identified and cloned from B. dorsalis. To our knowledge, the number of cecropins identified in the present study is greater than that reported in the insects of earlier studies. Moreover, some of the cecropins identified were significantly down-regulated after PS-1 treatment. CONCLUSION: Our findings provide new insights into the insect gut response to pathogenic bacterial invasion and may aid the development of new strategies for the biological control of B. dorsalis. © 2019 Society of Chemical Industry.

Biogenic amine biosynthetic and transduction genes in the endoparasitoid wasp Pteromalus puparum (Hymenoptera: Pteromalidae).

Biogenic amines (BAs), such as octopamine, tyramine, dopamine, serotonin, and acetylcholine regulate various behaviors and physiological functions in insects. Here, we identified seven genes encoding BA biosynthetic enzymes and 16 genes encoding BA G protein-coupled receptors in the genome of the endoparasitoid wasp, Pteromalus puparum. We compared the genes with their orthologs in its host Pieris rapae and the related ectoparasitic wasp Nasonia vitripennis. All the genes show high (>90%) identity to orthologs in N. vitripennis. P. puparum and N. vitripennis have the smallest number of BA receptor genes among the insect species we investigated. We then analyzed the expression profiles of the genes, finding those acting in BA biosynthesis were highly expressed in adults and larvae and those encoding BA receptors are highly expressed in adults than immatures. Octα1R and 5-HT7 genes were highly expressed in salivary glands, and a high messenger RNA level of 5-HT1A was found in venom apparatuses. We infer that BA signaling is a fundamental component of the organismal organization, homeostasis and operation in parasitoids, some of the smallest insects.

Genome-wide characterization and transcriptomic analyses of neuropeptides and their receptors in an endoparasitoid wasp, Pteromalus puparum.

In insects, neuropeptides constitute a group of signaling molecules that act in regulation of multiple physiological and behavioral processes by binding to their corresponding receptors. On the basis of the bioinformatic approaches, we screened the genomic and transcriptomic data of the parasitoid wasp, Pteromalus puparum, and annotated 36 neuropeptide precursor genes and 33 neuropeptide receptor genes. Compared to the number of precursor genes in Bombyx mori (Lepidoptera), Chilo suppressalis (Lepidoptera), Drosophila melanogaster (Diptera), Nilaparvata lugens (Hemiptera), Apis mellifera (Hymenoptera), and Tribolium castaneum (Coleoptera), P. puparum (Hymenoptera) has the lowest number of neuropeptide precursor genes. This lower number may relate to its parasitic life cycle. Transcriptomic data of embryos, larvae, pupae, adults, venom glands, salivary glands, ovaries, and the remaining carcass revealed stage-, sex-, and tissue-specific expression patterns of the neuropeptides, and their receptors. These data provided basic information about the identity and expression profiles of neuropeptides and their receptors that are required to functionally address their biological significance in an endoparasitoid wasp.

Identification, characterization and expression analysis of transient receptor potential channel genes in the oriental fruit fly, Bactrocera dorsalis.

BACKGROUND: Members of the transient receptor potential (TRP) superfamily are proteins that are critical for insects to detect changes in environmental stimuli and also play key roles in their sensory physiology. Moreover, this family provides potential targets for the design of insecticides. In contrast to a large number of studies conducted on Drosophila melanogaster, molecular studies to characterize TRP channels in agricultural pests are lacking. RESULTS: In this study, we identified 15 TRP channel genes in the genome of a notorious agricultural pest, the oriental fruit fly (Bactrocera dorsalis). Comparative analysis of the TRP channels (TRPs) in B. dorsalis with those in D. melanogaster, Glossina morsitans, Musca domestica and the closely related Ceratitis capitata, and TRPs from mosquitoes, Hymenoptera, Lepidoptera, Coleoptera and Hemiptera reveals that members of TRPA and TRPP subfamily are most diverse among insects. The results also suggest that Tephritidae family have two TRP-Polycystin 2 members even though most insects either possess just one or none. The highest expression levels of these two genes are in the testes of B. dorsalis, implying a role in regulating sperm function. We analyzed the expression profiles of the TRP channels identified in this study at different life stages using quantitative real time PCR. The results of this study demonstrate that all TRP channels are mainly expressed in adults, especially at mature stages. The one exception to this trend is BdTRPM, which is more highly expressed in the eggs of B. dorsalis, implying an important role in early development. We also detected the spatial expression of TRP channels in mature adult fruit flies by investigating expression levels within various tissues including those involved in sensory function, such as antennae, compound eyes, mouthparts, legs, and wings, as well as tissues critical for homeostasis and physiology (i.e., Malpighian tubules, the brain and gut as well as fat bodies, ovaries, and testes). CONCLUSION: The results of this study establish a solid foundation for future functional characterization of B. dorsalis TRP channels as well as those of other insects and will help future insecticide design targeting these channels.

Biogenic amine signaling systems in the red imported fire ant, Solenopsis invicta - Possible contributors to worker division of labor.

The red imported fire ant, Solenopsis invicta Buren, is a dangerous invasive pest in the United States, China and other countries. Efficient division of labor is one of the main reasons for the success of this social insect. Biogenic amines are important regulators of worker division of labor in this eusocial insect, but the related molecular mechanisms are largely unknown. In this study, we identified 10 candidate biogenic amine synthetic enzyme genes and 17 candidate biogenic amine receptor genes in the genome of S. invicta. Quantitative real-time PCR results indicated that foragers had higher head transcripts levels of all the tested enzyme genes than nurses did. In the abdomen, only the rate-limiting enzyme genes for the biosynthesis of serotonin and dopamine were higher in foragers than in nurses. Among the tested serotonin receptors, only the expression of 5-HT2A gene showed significant difference between foragers and nurses. In the head, more abundant 5-HT2A transcripts were detected in foragers than in nurses. Foragers expressed higher Octß4R than nurses in the head and abdomen. However, much lower mRNA levels of Dop3 receptor gene were detected in both body regions of foragers than nurses. Several other octopamine and tyramine receptor genes were also differentially expressed between foragers and nurses in the head and/or in the abdomen. Our results will improve the understanding of molecular mechanisms underlying biogenic amine modulation of the worker division of labor in S. invicta.

A new Drosophila octopamine receptor responds to serotonin.

As the counterparts of the vertebrate adrenergic transmitters, octopamine and tyramine are important physiological regulators in invertebrates. They control and modulate many physiological and behavioral functions in insects. In this study, we reported the pharmacological properties of a new α2-adrenergic-like octopamine receptor (CG18208) from Drosophila melanogaster, named DmOctα2R. This new receptor gene encodes two transcripts by alternative splicing. The long isoform DmOctα2R-L differs from the short isoform DmOctα2R-S by the presence of an additional 29 amino acids within the third intracellular loop. When heterologously expressed in mammalian cell lines, both receptors were activated by octopamine, tyramine, epinephrine and norepinephrine, resulting in the inhibition of cAMP production in a dose-dependent manner. The long form is more sensitive to the above ligands than the short form. The adrenergic agonists naphazoline, tolazoline and clonidine can stimulate DmOctα2R as full agonists. Surprisingly, serotonin and serotoninergic agonists can also activate DmOctα2R. Several tested adrenergic antagonists and serotonin antagonists blocked the action of octopamine or serotonin on DmOctα2R. The data presented here reported an adrenergic-like G protein-coupled receptor activated by serotonin, suggesting that the neurotransmission and neuromodulation in the nervous system could be more complex than previously thought.

Characterization of three serotonin receptors from the small white butterfly, Pieris rapae.

Serotonin (5-hydroxytryptamine, 5-HT) plays a key role in modulating diverse physiological processes and behaviors in both protostomes and deuterostomes. These functions are mediated through the binding of serotonin to its receptors, which are recognized as potential insecticide targets. We investigated the sequence, pharmacology and tissue distribution of three 5-HT receptors (Piera5-HT1A, Piera5-HT1B, Piera5-HT7) from the small white butterfly Pieris rapae, an important pest of cultivated cabbages and other mustard family crops. Activation of Piera5-HT1A or Piera5-HT1B by 5-HT inhibited the production of cAMP in a dose-dependent manner. Stimulation of Piera5-HT7 with 5-HT increased cAMP level significantly. Surprisingly, with the exception of 5-methoxytryptamine, agonists including α-methylserotonin, 8-Hydroxy-DPAT and 5-carboxamidotryptamine activated these receptors poorly. The results are consistent with previous findings in Manduca sexta. All three receptors were blocked by methiothepin, but ketanserin and yohimbine were not effective. The selective mammalian 5-HT receptor antagonists SB 216641 and SB 269970 displayed potent inhibition effects on Piera5-HT1B and Piera5-HT7 respectively. The results we achieved here indicate that the pharmacological properties of Lepidoptera 5-HT receptors are quite different from those in other insects and vertebrates and may contribute to development of new selective pesticides. This study offers important information on three 5-HT receptors from P. rapae that will facilitate further analysis of the functions of 5-HT receptors in insects.

Neuromodulation of Courtship Drive through Tyramine-Responsive Neurons in the Drosophila Brain.

Neuromodulators influence the activities of collections of neurons and have profound impacts on animal behavior. Male courtship drive is complex and subject to neuromodulatory control. Using the fruit fly Drosophila melanogaster, we identified neurons in the brain (inferior posterior slope; IPS) that impact courtship drive and were controlled by tyramine-a biogenic amine related to dopamine, whose roles in most animals are enigmatic. We knocked out a tyramine-specific receptor, TyrR, which was expressed in IPS neurons. Loss of TyrR led to a striking elevation in courtship activity between males. This effect occurred only in the absence of females, as TyrR(Gal4) mutant males exhibited a wild-type preference for females. Artificial hyperactivation of IPS neurons caused a large increase in male-male courtship, whereas suppression of IPS activity decreased male-female courtship. We conclude that TyrR is a receptor for tyramine, and suggest that it serves to curb high levels of courtship activity through functioning as an inhibitory neuromodulator.

Serotonin modulates insect hemocyte phagocytosis via two different serotonin receptors.

Serotonin (5-HT) modulates both neural and immune responses in vertebrates, but its role in insect immunity remains uncertain. We report that hemocytes in the caterpillar, Pieris rapae are able to synthesize 5-HT following activation by lipopolysaccharide. The inhibition of a serotonin-generating enzyme with either pharmacological blockade or RNAi knock-down impaired hemocyte phagocytosis. Biochemical and functional experiments showed that naive hemocytes primarily express 5-HT1B and 5-HT2B receptors. The blockade of 5-HT1B significantly reduced phagocytic ability; however, the blockade of 5-HT2B increased hemocyte phagocytosis. The 5-HT1B-null Drosophila melanogaster mutants showed higher mortality than controls when infected with bacteria, due to their decreased phagocytotic ability. Flies expressing 5-HT1B or 5-HT2B RNAi in hemocytes also showed similar sensitivity to infection. Combined, these data demonstrate that 5-HT mediates hemocyte phagocytosis through 5-HT1B and 5-HT2B receptors and serotonergic signaling performs critical modulatory functions in immune systems of animals separated by 500 million years of evolution.

Larvae of the small white butterfly, Pieris rapae, express a novel serotonin receptor.

The biogenic amine serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter in vertebrates and invertebrates. It acts in regulation and modulation of many physiological and behavioral processes through G-protein-coupled receptors. Five 5-HT receptor subtypes have been reported in Drosophila that share high similarity with mammalian 5-HT1A, 5-HT1B, 5-HT2A, 5-HT2B, and 5-HT7 receptors. We isolated a cDNA (Pr5-HT8 ) from larval Pieris rapae, which shares relatively low similarity to the known 5-HT receptor classes. After heterologous expression in HEK293 cells, Pr5-HT8 mediated increased [Ca(2+)]i in response to low concentrations (< 10 nM) of 5-HT. The receptor did not affect [cAMP]i even at high concentrations (> 10 µM) of 5-HT. Dopamine, octopamine, and tyramine did not influence receptor signaling. Pr5-HT8 was also activated by various 5-HT receptor agonists including 5-methoxytryptamine, (±)-8-Hydroxy-2-(dipropylamino) tetralin, and 5-carboxamidotryptamine. Methiothepin, a non-selective 5-HT receptor antagonist, activated Pr5-HT8 . WAY 10635, a 5-HT1A antagonist, but not SB-269970, SB-216641, or RS-127445, inhibited 5-HT-induced [Ca(2+)]i increases. We infer that Pr5-HT8 represents the first recognized member of a novel 5-HT receptor class with a unique pharmacological profile. We found orthologs of Pr5-HT8 in some insect pests and vectors such as beetles and mosquitoes, but not in the genomes of honeybee or parasitoid wasps. This is likely to be an invertebrate-specific receptor because there were no similar receptors in mammals. We isolated a cDNA (Pr5-HT8) from larval Pieris rapae, which shares relatively low similarity to the known GPCRs. After heterologous expression in HEK293 cells, Pr5-HT8 mediated increased [Ca(2+)]i in response to low concentrations (< 10 nM) of 5-HT and various 5-HT receptor agonists. We found orthologs of Pr5-HT8 in some insect pests and vectors such as beetles and mosquitoes, but not in the genomes of honeybee, parasitoid wasps, or mammals.

Two splicing variants of a novel family of octopamine receptors with different signaling properties.

The octopamine and tyramine, as the invertebrate counterparts of the vertebrate adrenergic transmitters, control and modulate many physiological and behavioral processes. Both molecules mediate their effects by binding to specific receptors belonging to the superfamily of G-protein-coupled receptors. So far, four families of octopamine and tyramine receptors have been reported. Here, we described the functional characterization of one putative octopamine/tyramine receptor gene from the rice stem borer, Chilo suppressalis. By a mechanism of alternative splicing, this receptor gene (CsOA3) encodes two molecularly distinct transcripts, CsOA3S and CsOA3L. CsOA3L differs from CsOA3S on account of the presence of an additional 30 amino acids within the third intracellular loop. When heterologously expressed, both receptors cause increases of intracellular Ca(2+) concentration. The short form, CsOA3S, was activated by both octopamine and tyramine, resulting in decreased intracellular cAMP levels ([cAMP]i ) in a dose-dependent manner, whereas dopamine and serotonin are not effective. However, CsOA3L did not show any impact on [cAMP]i . Studies with series of agonists and antagonists confirmed that CsOA3 has a different pharmacological profile from that of other octopamine receptor families. The CsOA3 is, to our knowledge, a novel family of insect octopamine receptors.

Parasitization by Cotesia chilonis influences gene expression in fatbody and hemocytes of Chilo suppressalis.

BACKGROUND: During oviposition many parasitoid wasps inject various factors, such as polydnaviruses (PDVs), along with eggs that manipulate the physiology and development of their hosts. These manipulations are thought to benefit the parasites. However, the detailed mechanisms of insect host-parasitoid interactions are not fully understood at the molecular level. Based on recent findings that some parasitoids influence gene expression in their hosts, we posed the hypothesis that parasitization by a braconid wasp, Cotesia chilonis, influences the expression of genes responsible for development, metabolism and immune functions in the fatbody and hemocytes of its host, Chilo suppressalis. METHODOLOGY/PRINCIPAL FINDINGS: We obtained 39,344,452 reads, which were assembled into 146,770 scaffolds, and 76,016 unigenes. Parasitization impacted gene expression in fatbody and hemocytes. Of these, 8096 fatbody or 5743 hemocyte unigenes were down-regulated, and 2572 fatbody or 1452 hemocyte unigenes were up-regulated. Gene ontology data showed that the majority of the differentially expressed genes are involved in enzyme-regulated activity, binding, transcription regulator activity and catalytic activity. qPCR results show that most anti-microbial peptide transcription levels were up-regulated after parasitization. Expression of bracovirus genes was detected in parasitized larvae with 19 unique sequences identified from six PDV gene families including ankyrin, CrV1 protein, cystatin, early-expressed (EP) proteins, lectin, and protein tyrosine phosphatase. CONCLUSIONS: The current study supports our hypothesis that parasitization influences the expression of fatbody and hemocyte genes in the host, C. suppressalis. The general view is that manipulation of host metabolism and immunity benefits the development and emergence of the parasitoid offsprings. The accepted beneficial mechanisms include the direct impact of parasitoid-associated virulence factors such as venom and polydnavirus on host tissues (such as cell damage) and, more deeply, the ability of these factors to influence gene expression. We infer that insect parasitoids generally manipulate their environments, the internal milieu of their hosts.