Toxoneuron Nigriceps Research Articles

An Overview of Ovarian Calyx Fluid Proteins of Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae): An Integrated Transcriptomic and Proteomic Approach.

The larval stages of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae), are parasitized by the endophagous parasitoid wasp, Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae). During the injections of eggs, this parasitoid wasp also injects into the host body the secretion of the venom gland and the calyx fluid, which contains a polydnavirus (T. nigriceps BracoVirus: TnBV) and the Ovarian calyx fluid Proteins (OPs). The effects of the OPs on the host immune system have recently been described. In particular, it has been demonstrated that the OPs cause hemocytes to undergo a number of changes, such as cellular oxidative stress, actin cytoskeleton modifications, vacuolization, and the inhibition of hemocyte encapsulation capacity, which results in both a loss of hemocyte functionality and cell death. In this study, by using a combined transcriptomic and proteomic analysis, the main components of T. nigriceps ovarian calyx fluid proteins were identified and their possible role in the parasitic syndrome was discussed. This study provides useful information to support the analysis of the function of ovarian calyx fluid proteins, to better understand T. nigriceps parasitization success and for a more thorough understanding of the components of ovarian calyx fluid proteins and their potential function in combination with other parasitoid factors.

Identification and Functional Characterization of Toxoneuron nigriceps Ovarian Proteins Involved in the Early Suppression of Host Immune Response.

Simple SummaryThe endoparasitoid of the tobacco budworm Heliothis virescens, Toxoneuron nigriceps, has several strategies to survive, including venom and calyx fluid. This latter contains a Polydnavirus and Ovarian Proteins (OPs). They are injected into the host body together with the egg. Although much research has focused on venom protein components, little is known about OPs. OPs can disrupt the cellular immune response of the host, acting on host hemocytes, the immune cells. In this study we investigated the action of HPLC fractions derived from OPs. Two fractions caused multiple and significant changes in hemocytes, including cellular oxidative stress and actin cytoskeleton disruption, which might explain the high incidence of hemocyte death and loss of function. Furthermore, using a transcriptome and proteomic approach, we identify the proteins of the two fractions that may be involved in the observed host hemocyte alterations. Our results will help to better understand the OP components and their involvement in parasitization strategies.The endophagous parasitoid Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) of the larval stages of the tobacco budworm Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae) injects the egg, the venom, the calyx fluid, which includes a Polydnavirus (T. nigriceps BracoVirus: TnBV) and the Ovarian Proteins (OPs) into the host body during oviposition. The host metabolism and immune system are disrupted prematurely shortly after parasitization by the combined action of the TnBV, venom, and OPs. OPs are involved in the early suppression of host immune response, before TnBV infects and expresses its genes in the host tissues. In this work, we evaluated the effect of HPLC fractions deriving from in toto OPs. Two fractions caused a reduction in hemocyte viability and were subsequently tested to detect changes in hemocyte morphology and functionality. The two fractions provoked severe oxidative stress and actin cytoskeleton disruption, which might explain the high rate of hemocyte mortality, loss of hemocyte functioning, and hence the host’s reduced hemocyte encapsulation ability. Moreover, through a transcriptome and proteomic approach we identify the proteins of the two fractions: eight proteins were identified that might be involved in the observed host hemocyte changes. Our findings will contribute to a better understanding of the secreted ovarian components and their role in parasitoid wasp strategy for evading host immune responses.

Role of Ovarian Proteins Secreted by Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) in the Early Suppression of Host Immune Response.

Simple SummaryToxoneuron nigriceps is an endoparasitoid of the tobacco budworm Heliothis virescens. Parasitoid strategies to survive involve different regulating factors that are injected into the host body together with the egg: the venom and the calyx fluid, containing a Polydnavirus (PDV) and Ovarian Proteins (OPs). The combination of these factors increases the success of parasitism. Although many studies have been reported on venom protein components and the knowledge on PDVs is increasing, little is known on OPs. These secretions are able to interfere early with the host cellular immune response, acting specifically on host haemocytes, cells involved in immune response. Our results show that OPs induce several alterations on haemocytes, including cellular oxidative stress condition and modifications of actin cytoskeleton, so inducing both a loss of haemocyte functionality and cell death. Overall, in synergy with PDV and venom, OPs positively contribute to the evasion of the host immune response by T. nigriceps.Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) is an endophagous parasitoid of the larval stages of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae). During oviposition, T. nigriceps injects into the host body, along with the egg, the venom, the calyx fluid, which contains a Polydnavirus (T. nigriceps BracoVirus: TnBV), and the Ovarian Proteins (OPs). Although viral gene expression in the host reaches detectable levels after a few hours, a precocious disruption of the host metabolism and immune system is observed right after parasitization. This alteration appears to be induced by female secretions including TnBV venom and OPs. OPs, originating from the ovarian calyx cells, are involved in the induction of precocious symptoms in the host immune system alteration. It is known that OPs in braconid and ichneumonid wasps can interfere with the cellular immune response before Polydnavirus infects and expresses its genes in the host tissues. Here we show that T. nigriceps OPs induce several alterations on host haemocytes that trigger cell death. The OP injection induces an extensive oxidative stress and a disorganization of actin cytoskeleton and these alterations can explain the high-level of haemocyte mortality, the loss of haemocyte functionality, and so the reduction in encapsulation ability by the host.

Beyond host specificity: the biotechnological exploitation of chitolectin from teratocytes of Toxoneuron nigriceps to control non-permissive hosts

The virulence factors (VF) that parasitoids use to regulate the growth and development of their hosts have potential as new events in transgenic plants. VFs are promising candidates for biotechnological exploitation and implementation of pest-control tools to meet the world’s demand for food in 2100, while sustainably increasing agricultural production. However, VFs have not been used in commercial transgenic plants, since they are thought to target only a narrow range of insects, due to the parasitoid host specialization. We investigated the potential of chitolectin (TnChit), produced by the teratocytes of Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae), as a transgenic event to control several non-permissive hosts: Chrysodeixis includens (Walker), Spodoptera albula (Walker), Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) and Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). Third instars of C. includens, S. albula and S. frugiperda had lower survival and probability of molting on TnChit-modified plants than on control plants. The nutritional indices of S. frugiperda were also altered. Larval survival and pupal weight of males of T. absoluta were reduced on transgenic plants. Adult emergence of B. tabaci was also reduced. The expression of the TnChit gene differed in the transformation events obtained and was consistent with the biological effects observed. The effects of TnChit on non-permissive hosts demonstrate the potential of the virulence factors that parasitoids employ in host regulation, as a source of new genes in plant transformation for control of insect pests.

Novel Factors of Viral Origin Inhibit TOR Pathway Gene Expression.

Polydnaviruses (PDVs) are obligate symbionts of endoparasitoid wasps, which exclusively attack the larval stages of their lepidopteran hosts. The Polydnavirus is injected by the parasitoid female during oviposition to selectively infect host tissues by the expression of viral genes without undergoing replication. Toxoneuron nigriceps bracovirus (TnBV) is associated with Toxoneuron nigriceps (Hymenoptera: Braconidae) wasp, an endoparasitoid of the tobacco budworm larval stages, Heliothis virescens (Lepidoptera: Noctuidae). Previous studies showed that TnBV is responsible for alterations in host physiology. The arrest of ecdysteroidogenesis is the main alteration which occurs in last (fifth) instar larvae and, as a consequence, prevents pupation. TnBV induces the functional inactivation of H. virescens prothoracic glands (PGs), resulting in decreased protein synthesis and phosphorylation. Previous work showed the involvement of the PI3K/Akt/TOR pathway in H. virescens PG ecdysteroidogenesis. Here, we demonstrate that this cellular signaling is one of the targets of TnBV infection. Western blot analysis and enzyme immunoassay (EIA) showed that parasitism inhibits ecdysteroidogenesis and the phosphorylation of the two targets of TOR (4E-BP and S6K), despite the stimulation of PTTH contained in the brain extract. Using a transcriptomic approach, we identified viral genes selectively expressed in last instar H. virescens PGs, 48 h after parasitization, and evaluated expression levels of PI3K/Akt/TOR pathway genes in these tissues. The relative expression of selected genes belonging to the TOR pathway (tor, 4e-bp, and s6k) in PGs of parasitized larvae was further confirmed by qRT-PCR. The down-regulation of these genes in PGs of parasitized larvae supports the hypothesis of TnBV involvement in blocking ecdysteroidogenesis, through alterations of the PI3K/Akt/TOR pathway at the transcriptional level.

The mechanism utilized by Toxoneuron nigriceps in inhibiting the host immune system

Insect parasitic factors of both maternal and embryonic origin allow the endoparasitoid progeny to elude the humoral and cellular immune system responses of their host. Endoparasitoid wasps of lepidopteran larval stages inject, along with venom and ovarian proteins, Polydnavirus particles acting in synergy with all other factors in host regulation for parasitism success. To date, the molecular mechanisms used by endoparasitoid to circumvent the host immune system are little known. Neverthless, several of these strategies are conserved through the wasp parasitoid species. Heliothis virescens is a noctuid moth, host of the endophagous parasitoid Toxoneuron nigriceps. The first observed effect of parasitism is immune system suppression, as direct consequence of the array of host regulation factors, both of embryonic and maternal origin. This review describes the contribution of all the parasitic components during alterations of the host immune response observed after oviposition by T. nigriceps.

A polydnavirus-encoded ANK protein has a negative impact on steroidogenesis and development

Polydnaviruses (PDV) are viral symbionts associated with ichneumonid and braconid wasps parasitizing moth larvae, which are able to disrupt the host immune response and development, as well as a number of other physiological pathways. The immunosuppressive role of PDV has been more intensely investigated, while very little is known about the PDV-encoded factors disrupting host development. Here we address this research issue by further expanding the functional analysis of ankyrin genes encoded by the bracovirus associated with Toxoneuron nigriceps (Hymenoptera, Braconidae). In a previous study, using Drosophila melanogaster as experimental model system, we demonstrated the negative impact of TnBVank1 impairing the ecdysone biosynthesis by altering endocytic traffic in prothoracic gland cells. With a similar approach here we demonstrate that another member of the viral ank gene family, TnBVank3, does also contribute to the disruption of ecdysone biosynthesis, but with a completely different mechanism. We show that its expression in Drosophila prothoracic gland (PG) blocks the larval-pupal transition by impairing the expression of steroidogenic genes. Furthermore, we found that TnBVank3 affects the expression of genes involved in the insulin/TOR signaling and the constitutive activation of the insulin pathway in the PG rescues the pupariation impairment. Collectively, our data demonstrate that TnBVANK3 acts as a virulence factor by exerting a synergistic and non-overlapping function with TnBVANK1 to disrupt the ecdysone biosynthesis.

The multifunctional polydnavirus TnBVANK1 protein: impact on host apoptotic pathway

Toxoneuron nigriceps (Hymenoptera, Braconidae) is an endophagous parasitoid of the larval stages of the tobacco budworm, Heliothis virescens (Lepidoptera, Noctuidae). The bracovirus associated with this wasp (TnBV) is currently being studied. Several genes expressed in parasitised host larvae have been isolated and their possible roles partly elucidated. TnBVank1 encodes an ankyrin motif protein similar to insect and mammalian IκB, an inhibitor of the transcription nuclear factor κB (NF-κB). Here we show that, when TnBVank1 was stably expressed in polyclonal Drosophila S2 cells, apoptosis is induced. Furthermore, we observed the same effects in haemocytes of H. virescens larvae, after TnBVank1 in vivo transient transfection, and in haemocytes of parasitised larvae. Coimmunoprecipitation experiments showed that TnBVANK1 binds to ALG-2 interacting protein X (Alix/AIP1), an interactor of apoptosis-linked gene protein 2 (ALG-2). Using double-immunofluorescence labeling, we observed the potential colocalization of TnBVANK1 and Alix proteins in the cytoplasm of polyclonal S2 cells. When Alix was silenced by RNA interference, TnBVANK1 was no longer able to cause apoptosis in both S2 cells and H. virescens haemocytes. Collectively, these results indicate that TnBVANK1 induces apoptosis by interacting with Alix, suggesting a role of TnBVANK1 in the suppression of host immune response observed after parasitisation by T. nigriceps.

Identification of major Toxoneuron nigriceps venom proteins using an integrated transcriptomic/proteomic approach

Endoparasitoids in the order Hymenoptera are natural enemies of several herbivorous insect pest species. During oviposition they inject a mixture of factors, which include venom, into the host, ensuring the successful parasitism and the development of their progeny. Although these parasitoid factors are known to be responsible for host manipulation, such as immune system suppression, little is known about both identity and function of the majority of their venom components. To identify the major proteins of Toxoneuron nigriceps (Hymenoptera: Braconidae) venom, we used an integrated transcriptomic and proteomic approach. The tandem-mass spectrometric (LC-MS/MS) data combined with T. nigriceps venom gland transcriptome used as a reference database resulted in the identification of a total of thirty one different proteins. While some of the identified proteins have been described in venom from several parasitoids, others were identified for the first time. Among the identified proteins, hydrolases constituted the most abundant family followed by transferases, oxidoreductases, ligases, lyases and isomerases. The hydrolases identified in the T. nigriceps venom glands included proteases, peptidases and glycosidases, reported as common components of venom from several parasitoid species. Taken together, the identified proteins included factors that could potentially inhibit the host immune system, manipulate host physiological processes and host development, as well as provide nutrients to the parasitoid progeny, degrading host tissues by specific hydrolytic enzymes.The venom decoding provides us with information about the identity of candidate venom factors which could contribute to the success of parasitism, together with other maternal and embryonic factors.

Systemic Imidacloprid Affects Intraguild Parasitoids Differently.

Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) and Campoletis sonorensis (Cameron) (Hymenoptera, Ichneumonidae) are solitary endoparasitoids of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae). They provide biological control of H. virescens populations in Southeastern US agricultural production systems. Field and greenhouse experiments conducted from 2011–2014 compared parasitism rates of parasitoids that developed inside H. virescens larvae fed on tobacco plants treated with and without imidacloprid. The parasitoids in our study did not have a similar response. Toxoneuron nigriceps had reduced parasitism rates, but parasitism rates of C. sonorensis were unaffected. Preliminary data indicate that adult female lifespans of T. nigriceps are also reduced. ELISA was used to measure concentrations of neonicotinoids, imidacloprid and imidacloprid metabolites in H. virescens larvae that fed on imidacloprid-treated plants and in the parasitoids that fed on these larvae. Concentrations were detectable in the whole bodies of parasitized H. virescens larvae, T. nigriceps larvae and T. nigriceps adults, but not in C. sonorensis larvae and adults. These findings suggest that there are effects of imidacloprid on multiple trophic levels, and that insecticide use may differentially affect natural enemies with similar feeding niches.

Postegression Feeding Enhances Growth, Survival, and Nutrient Acquisition in the Endoparasitoid Toxoneuron nigriceps (Hymenoptera: Braconidae).

Toxoneuron nigriceps Viereck (Hymenoptera: Braconidae), a koinobiont endoparasitoid of the tobacco budworm, Heliothis virescens F. (Lepidoptera: Noctuidae), derives nutrition from the host hemolymph during the internal portion of its larval development but feeds destructively on host tissues externally after egression. To investigate the importance of this tissue-feeding phase, and to evaluate the behaviors associated with postegression feeding, T. nigriceps larvae were subjected to one of four treatments: 1) allowed to carry out normal tissue feeding, 2) deprived of tissue feeding, 3) presented with tissues scraped away from the host remains, and 4) fed tissues scraped from an unparasitized H. virescens larva. Additionally, total carbohydrates, lipids, and proteins were quantified from pre and posttissue feeding T. nigriceps larvae to examine the effect of postegression feeding on parasitoid nutritional physiology. Parasitoids that received no tissues after egression, or that received tissue from an unparasitized H. virescens larva, had significantly smaller body masses at all stages than those allowed to feed naturally or fed tissues scraped from a parasitized host. Parasitoids that underwent normal host feeding after egression also reached larger masses then those fed scraped host tissue. Parasitoids that received no tissue after egression survived to adulthood significantly less often than those that were presented with any H. virescens tissue. This suggests that postegression tissue feeding is a vital developmental step for T. nigriceps, and that T. nigriceps will not only feed when normal postegression behavior is disrupted, but will also feed on unparasitized tissue. The quantification of macronutrients in the tissues of pre and posttissue feeding T. nigriceps larvae showed significantly elevated proportions of proteins, lipids, and carbohydrates in the tissues of larvae that had completed feeding, with the greatest difference being in total lipids.

A virulence factor encoded by a polydnavirus confers tolerance to transgenic tobacco plants against lepidopteran larvae, by impairing nutrient absorption.

The biological control of insect pests is based on the use of natural enemies. However, the growing information on the molecular mechanisms underpinning the interactions between insects and their natural antagonists can be exploited to develop “bio-inspired” pest control strategies, mimicking suppression mechanisms shaped by long co-evolutionary processes. Here we focus on a virulence factor encoded by the polydnavirus associated with the braconid wasp Toxoneuron nigriceps (TnBV), an endophagous parasitoid of noctuid moth larvae. This virulence factor (TnBVANK1) is a member of the viral ankyrin (ANK) protein family, and appears to be involved both in immunosuppression and endocrine alterations of the host. Transgenic tobacco plants expressing TnBVANK1 showed insecticide activity and caused developmental delay in Spodoptera littoralis larvae feeding on them. This effect was more evident in a transgenic line showing a higher number of transcripts of the viral gene. However, this effect was not associated with evidence of translocation into the haemocoel of the entire protein, where the receptors of TnBVANK1 are putatively located. Indeed, immunolocalization experiments evidenced the accumulation of this viral protein in the midgut, where it formed a thick layer coating the brush border of epithelial cells. In vitro transport experiments demonstrated that the presence of recombinant TnBVANK1 exerted a dose-dependent negative impact on amino acid transport. These results open new perspectives for insect control and stimulate additional research efforts to pursue the development of novel bioinsecticides, encoded by parasitoid-derived genes. However, future work will have to carefully evaluate any effect that these molecules may have on beneficial insects and on non-target organisms.

A polydnavirus ANK protein acts as virulence factor by disrupting the function of prothoracic gland steroidogenic cells.

Polydnaviruses are obligate symbionts integrated as proviruses in the genome of some ichneumonoid wasps that parasitize lepidopteran larvae. Polydnavirus free viral particles, which are injected into the host at oviposition, express virulence factors that impair immunity and development. To date, most studies have focused on the molecular mechanisms underpinning immunosuppression, whereas how viral genes disrupt the endocrine balance remains largely uninvestigated. Using Drosophila as a model system, the present report analyzes the function of a member of the ankyrin gene family of the bracovirus associated with Toxoneuron nigriceps, a larval parasitoid of the noctuid moth Heliothis virescens. We found that the TnBVank1 expression in the Drosophila prothoracic gland blocks the larval-pupal molt. This phenotype can be rescued by feeding the larvae with 20-hydroxyecdysone. The localization of the TnBVANK1 is restricted to the cytoplasm where it interacts with Hrs and Alix marked endosomes. Collectively, our data demonstrate that the TnBVANK1 protein acts as a virulence factor that causes the disruption of ecdysone biosynthesis and developmental arrest by impairing the vesicular traffic of ecdysteroid precursors in the prothoracic gland steroidogenic cells.

Characterization of a Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) – derived chitinase and its potential for pest control

The larval endoparasitoid Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) has a toolbox of biological weapons to secure for host colonization and the successful parasitization of its host Heliothis virescens (F.) (Lepidoptera: Noctuidae). The cDNA of a putative chitinase has been previously isolated and initially characterized from teratocytes of this parasitoid among the plethora of molecules available in the venom and calyx fluids injected by females, oral and/or anal secretions released by the parasitoid larvae and/or produced by the expression of genes of the symbiotic associated polydnavirus. This putative chitinase has been initially associated with the host cuticle digestion to allow for parasitoid egression and with the asepsis of the host environment, acting as an antimicrobial. As chitinases are commonly expressed in plants against plant pathogens, the chitinase derived from the teratocytes of T. nigriceps is a potential tool for the development of insect pest control methods based on the disruption of the perithrophic membrane of herbivores. Therefore, we aimed to characterize the activity of the putative chitinase from teratocytes of T. nigriceps (Tnchi) produced using the Escherichia coli expression system and its potential to control H. virescens larvae when expressed into transgenic tobacco plants. The purified E. coli-produced Tnchi protein showed no chitinolitic activity, but was active in binding with colloidal and crystalline chitins in water and with colloidal chitin in buffered solution (pH=6.74). Transgenic tobacco plants showed no enhanced chitinolitic activity relative to control plants, but survival of three-day old larvae of H. virescens was severely affected when directly fed on transgenic tobacco leaves expressing the recombinant Tnchi protein. Some properties of the Tnchi protein and the potential use of Tnchi-transgenic plants to control plant pests are discussed.

Suitability of Various Artificial Pupation Chambers for use by Koinobiont Parasitoid Toxoneuron nigriceps (Hymenoptera: Braconidae)

Abstract Although koinobiont parasitoids depend on their hosts for nutrition, they often exploit them in other ways during pupation. The solitary endoparasitoid Toxoneuron nigriceps Viereck (Hymenoptera: Braconidae) spins its cocoon in a chamber formed by its host Heliothis virescens (F.) (Lepidoptera: Noctudidae). The parasitoid prepupae depend on this host-formed pupation chamber; if left on an open surface, T. nigriceps larvae are unable to form a cocoon. To determine whether T. nigriceps larvae can form cocoons in artificial pupation chambers, common laboratory materials were tested for suitability. Test chambers included cotton balls, upright test tubes, gelatin capsules, and the wells of microtube racks. The proportion of parasitoids that formed cocoons in each chamber, as well as the proportion that survived to adulthood, were recorded and compared with each other and to those placed in host-formed chambers, as well as to those placed in petri dishes, a flat surface. Parasitoid larvae were able to spin cocoons in all artificial chambers that provided a small, partially enclosed space. The proportion of parasitoids that formed cocoons was largest in cotton balls, but no chambers (other than petri dishes) were significantly different from the host formed control. The proportion of parasitoids that survived to adulthood was significantly smaller in the petri dish and test tube treatments than in the control. Due to the high success rate for cocoon formation and survival, as well as practicality in a mass-rearing situation, gelatin capsules and microtube rack wells were considered the best choices for use as artificial pupation chambers among those tested.

Post-Egression Host Tissue Feeding is Another Strategy of Host Regulation by the Koinobiont Wasp,Toxoneuron nigriceps

Koinobiont wasps start their lives as hemolymph feeders inside the host body, but before they egress from the host many become tissue predators. One species, the endoparasitoid Toxoneuron nigriceps Viereck (Hymenoptera: Braconidae), exhibits the unusual behavior of egressing before initiating tissue predation. After egression from the host, it reinserts its head into the host body to begin tissue feeding. These third instar T. nigriceps larvae show a significant increase in body size and mass after post-egression feeding. Through this project the importance of post-egression feeding in the development of T. nigriceps in its host the tobacco budworm, Heliothis virescens Fabricius (Lepidoptera: Noctuidae), has been evaluated. The study was conducted by preventing the egressed third instar T. nigriceps larvae from feeding on host tissue and observing whether they could undergo further development. Though some of the larvae that were prevented from post-egression feeding did undergo cocoon formation, pupation, and adult emergence they were inferior in terms of size, body mass, and survival to those that developed from larvae allowed to feeding after egression. Hence, it is concluded that post-egression host tissue feeding is essential for the normal development of T. nigriceps, as the prevention of feeding resulted in significantly lighter and smaller larvae, cocoons, and adults as well as deformed adults and reduced adult survival. Post-egression feeding as a host regulatory strategy is discussed.

The probable significance of tracheal tufts in the 8th abdominal segment of Heliothis virescens (F.) on the development of its parasitoid, Toxoneuron nigriceps (Viereck)

The 8th abdominal segment of Heliothis virescens (Fabricius) larvae contains aerating trachea and tracheole tufts that end in the hemocoel of the 8th segment, unlike the tracheae that invade tissues in other segments. These tracheal tufts from the 8th abdominal segment extend to the tokus region, which along with the telson cavity is known to act as a “lung” for hemocytes in Calpodes ethlius and a few other lepidopteran larvae. The goal of this research was to study the effects of these tracheal tufts in the 8th abdominal segment on parasitoid development inside the host larvae, H. virescens. The first objective was to determine if the eggs of the parasitoid, Toxoneuron nigriceps, are predominantly located among the tracheal tufts of the 8th abdominal segment compared to other body cavity regions irrespective of their oviposition site or the position of the host larvae. The results showed that several hours after oviposition most of the eggs are found in the 8th abdominal segment irrespective of the oviposition site or the position of the host larvae. The second objective was to study the effect of varying oxygen concentrations in vitro on various developmental stages of the egg. The results showed that decreasing oxygen concentrations adversely affects the parasitoid egg development in vitro. A third objective was to determine the oxygen concentration in 8th abdominal segment of the host larvae and compare it to other regions of the body using an oxygen sensor placed in vivo. The results suggested relatively high concentration of oxygen in the 8th abdominal segment compared to other regions of the host, thus supporting our hypothesis that the increased oxygen level in the 8th abdominal segment is important to the development of the parasitoid eggs.

Pest Insects and Natural Enemies in Transitional Organic Cotton in Georgia

This research was conducted in experimental plots in Georgia from 2004 through 2006 and assessed the prospects for transitioning to a totally organic management system for cotton. The seasonal abundance of insect pests and their natural enemies over the 3-yr transitional period are reported herein. The heliothines, Heliothis virescens (F.) and Helicoverpa zea (Boddie), and stink bugs, mainly Nezara viridula (L.), Euschistus servus (Say), and Euschistus quadrator (Rolston), were the 2 groups of insect pests found on cotton. Heliothine larvae were observed each year of the study but, in general, infestations of heliothines were higher in 2004 than in the other 2 yrs. Stink bugs were observed in relatively high numbers, and the percent of cotton bolls damaged by these pests was high only during year 2 of the study. Over the 3-yr period, the red imported fire ant, Solenopsis invicta Buren, various spiders, including Oxyopes scalaris Hentz and Peucetia viridans (Hentz), the big-eyed bug, Geocoris punctipes (Say), and the pirate bug, Orius insidiosus (Say), were the most abundant predators of the heliothines and stink bugs. The endoparasitoid Toxoneuron nigriceps Viereck and an ascovirus also contributed to larval mortality of H. virescens. The endoparasitoid Trichopoda pennipes (F.) parasitized adults of N. viridula.

Toxoneuron nigriceps parasitization delays midgut replacement in fifth-instar Heliothis virescens larvae

We have analyzed the effects of Toxoneuron nigriceps parasitization on the midgut development of its host Heliothis virescens. In parasitized H. virescens larvae, the midgut epithelium undergoes a complete replacement, which is qualitatively not different to that observed in synchronous unparasitized larvae, with similar temporal profiles of cell death and metabolic activity. However, the whole gut replacement process is significantly delayed in parasitized larvae, with complete differentiation of the new gut epithelium being observed 4 days later than in unparasitized controls. The administration of juvenile hormone before commitment and of 20-hydroxyecdysone (20E) after commitment delays and fosters, respectively, the replacement process of the midgut epithelium; moreover, the injection of 20E into developmentally arrested and 20E-deficient host last-instar larvae parasitized by T. nigriceps immediately triggers regular gut development. These hormone-based experiments suggest that endocrine alterations in the larval host, induced by T. nigriceps parasitism, are responsible for the temporal alterations in the gut replacement process. The role of this parasitoid-induced developmental change in the host regulation process is discussed.

Characterization of a cDNA encoding a putative chitinase from teratocytes of the endoparasitoidToxoneuron nigriceps

AbstractFemales of insect endoparasitoids inject substances along with the egg at oviposition, which can regulate or induce changes in a number of host physiological processes to benefit the developing parasitoid. These changes can be caused by substances such as venoms, calyx fluids, or symbiotic‐associated virus particles (polydnavirus), which are injected by females along with their eggs, and by substances secreted by parasitoid‐derived tissues (teratocytes) or the developing parasitoid larvae. Teratocytes (dissociated cells of the serosal membrane after parasitoid eclosion) release substances that have roles (i) in parasitoid nutrition, (ii) in the digestion of host tissues, and (iii) in the regulation of host development. Teratocytes ofToxoneuron nigriceps(Viereck) (Hymenoptera: Braconidae) have been implicated in the arrestment of the host development and in the regulation of circulating levels of host ecdysteroids. Here we describe the cDNA of a teratocyte‐secreted chitinase and its expression during parasitoid development, and discuss its putative role in this host–parasitoid association.