Microplitis croceipes teratocytes: in vitro culture and biological activity of teratocyte secreted protein

Teratocytes originate from the dissociation of the extraembryonic serosal membrane in some Braconidae and Scelionidae. Methods used to culture teratocytes in vitro are described and the yield of teratocyte secreted proteins (TSP) was measured. Although 90% are viable after 6 days, in vitro teratocytes reached only half the diameter (32&mgr;m) of the same age teratocytes obtained in vivo. Teratocytes cultured in vitro secrete as much as 0.7&mgr;g of protein per day per larval equivalent ( approximately 900 cells). Presence of parasitoid larvae enhanced teratocyte viability while periodic exchange of medium did not. However, medium exchange significantly increased the total amount of protein secreted. Size and viability were improved with the addition of 10% FBS to the Ex-cell 400 culture medium. Non-denaturing PAGE showed at least 15 proteins with molecular sizes estimated to be between 24 to 347kDa in medium containing teratocytes. An in vitro fat body assay was developed to measure the effect of TSP on protein synthesis and juvenile hormone esterase (JHE) activity. Crude TSP inhibited in vitro incorporation of [(35)S]-methionine into protein synthesized by the fat body. The amount of JHE released from in vitro fat body treated with crude TSP was significantly less than controls, most likely caused by the inhibition of general protein synthesis. The active fraction of TSP passed through a 30kDa molecular weight cutoff filter but was retained by a 3kDa filter. SDS-PAGE revealed four proteins with molecular weights between 8 and 20kDa not present in control medium incubated without teratocytes.     

Host regulation effects on Heliothis virescens (F.) larvae inducd by teratocytes of Cardiochiles nigriceps Viereck (Lepidoptera,Noctuidae-Hymenoptera,Braconidae)

Teratocytes deriving from the serosal membrane of Cardiochiles nigriceps Viereck, obtained “in vitro” from embryos hatched on a semidefined medium, were injected at different numbers and in different developmental stages of nonparasitized Heliothis virescens (F.) last instar larvae. Host development was affected by teratocyte injections and the responses registered ranged from normal to complete inhibition of pupation, according to the number of teratocytes injected and the developmental stage of the larva at time of injection. Complete pupation failure was observed when teratocytes derived from 4C nigriceps embryos were injected into 1st day 5th instar (new-slender stage) host larvae. Complete pupation occurred when teratocytes from 2 embryos were injected into 3rd or 4th day 5th instars (burrow-digging or day 1 cell formation stage). Intermediate responses, such as the formation of pupal cuticle without ecdysis or with only partial ecdysis, were obtained with intermediate teratocyte numbers, or host developmental stages. All pupae derived from teratocyte injected larvae failed to develop into adults normally obtained from control injected larvae. The larval weight just before pupation was negatively affected only when teratocyte injections were performed on 1st day 5th instar H. virescens larvae. Teratocyte injections altered the hemolymph protein titer to a level similar to that occurring in parasitized larvae. At the same time the ecdysteroid titer was characterized by a late significant increase, which reached values almost 3 times greater than found in normally parasitized larvae, and also surpassed the highest values registered for nonparasitized larvae. Ligation of parasitized larvae between the meso- and metathorax demonstrated that when the prothoracic glands were excluded, there was almost no ecdysteroid production posterior to the ligation. Ligations performed on parasitized larvae to isolate parasitoid eggs before hatching in the last abdominal segments, demonstrated that only virus and venom determined a reduction of the ecdysteroid titer. On the basis of these results the possible role of teratocytes in affecting the biological activity of ecdysteroids is postulated and discussed in a wider context of host-parasitoid physiological interactions.     

Dinocampus (=Perilitus) coccinellae teratocyte-specific polypeptide: its accumulative property,localization and characterization

Dinocampus (=Perilitus) coccinellae (Braconidae: Hymenoptera) teratocytes synthesize a teratocyte-specific polypeptide (TSP) with a high molecular weight of 540kDa. The TSP has a tendency to accumulate in the teratocyte cells without release after synthesis ([Okuda and Kadono-Okuda, 1995]), which was confirmed in this study. Pulse-chase fluorography indicated that teratocytes at a younger stage (6 days after parasitization)secreted negligible TSP into the medium after synthesis, while teratocytes at an older stage (11 days after parasitization)secreted the synthesized products into the medium, although the amount released was still low. Western blot with anti-TSP serum showed that only a small amount of TSP appeared in the parasitized host hemolymph, even when TSP synthesis by teratocytes was actively taking place, which also supported the accumulative nature of TSP. The immunoelectronmicroscopic studies revealed that the TSP was localized specifically in high electron-dense vacuoles. Lectin blot analysis identified TSP as a high mannose glycoprotein. The amino acid composition of the major subunit of the TSP was quite similar to that of nutritive proteins such as vitellogenin and storage proteins of some insects. These characterization data, together with the accumulation property of the TSP indicates that Dinocampus teratocyte primarily plays a nutritive role for the developing parasitoid larvae. TSP exhibited esterase activity, which indicates that TSP may have an additional function in the host-parasitoid reaction.     

Characterization of a cDNA encoding a putative chitinase from teratocytes of the endoparasitoid Toxoneuron nigriceps

Females 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 of Toxoneuron 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.     

Pea aphid clonal resistance to the endophagous parasitoid Aphidius ervi

The physiological mechanism of resistance to the endophagous braconid Aphidius ervi Haliday (Hymenoptera, Braconidae) by a pink clone (PC) of Acyrthosiphon pisum (Harris) (Homoptera, Aphididae) has been investigated. Comparative data on parasitoid development and associated host biochemical changes in the resistant PC aphids and in a susceptible green clone (GC) of A. pisum are reported. When the PC aphids were attacked as early 4th instars, the developing parasitoid larvae showed a strongly reduced increase in size, compared to those synchronously developing in GC aphids, and were unable to produce a regular mummy. In contrast, parasitism of 2nd instar PC aphids, allowed completion of parasitoid development, but adults had a prolonged developmental time, due to a longer duration of parasitoid’s final (3rd) instar. In all cases, teratocytes, cells deriving from the A. ervi serosal membrane, and the proteins abundantly synthesised by them, were never found in the haemolymph of parasitised PC aphids. Host castration, as demonstrated by total protein incorporation into reproductive tissues, was total in the majority of early (2nd instar) parasitised host aphids, while it was limited when later instars (4th) of PC aphids were parasitised. This is partly due to the absence of the cytolytic activity of teratocytes on host embryos, which, through their persistence, may compete for nutritional resources with the developing parasitoid larvae. In parasitised PC aphids, this competitive effect is further aggravated for the parasitoid by the absence of the regulated amino acid titre increase in the host haemolymph, which is regularly observed in GC aphids. Failure of teratocyte development in the PC clone of the pea aphid is, then, the major functional constraint accounting for the reduction/inhibition of A. ervi larval growth. The reported results allow to assess in vivo the role of teratocytes in the host physiological redirection and nutritional exploitation by the parasitoid, and to integrate and validate the proposed physiological model of host-parasitoid interactions in the system A. pisum-A.ervi.     

Cultivation of teratocytes of the egg parasitoid telenomus heliothidis (hymenoptera: Scelionidae)

Summary A method for in vitro cultivation of teratocytes from the egg parasitoidTelenomus heliothidis (Hymenoptera: Scelionidae) is described. Parasitoid eggs, from which teratocytes were derived, were collected from 24-h-oldHeliothis virescens (Lepidoptera: Noctuidae) eggs previously parasitized byT. heliothidis females. Optimal culture conditions, including species and concentration of serum, were determined experimentally. Thirty percentManduca sexta hemolymph or 10% chicken serum in Hink’s TNH-FH medium were found to generate the most satisfactory number of teratocytes per parasitoid larva. Teratocytes cultivated in vitro showed similar development and morphology to those produced in vivo. However, cultured teratocytes lived approximately 10 times longer than teratocytes in natural hosts and were not dependent upon the presence of the parasitoid larva for normal development.     

Development of teratocytes associated with Microctonus aethiopoides Loan (Hymenoptera: Braconidae) in natural and novel host species

A laboratory study investigated development of teratocytes derived from the parasitoid Microctonus aethiopoides Loan in the natural host, Sitona discoideus Gyllenhal, and in three novel hosts, the introduced weed biological control agent Rhinocyllus conicus (Froehlich), and two New Zealand native species Nicaeana cervina Broun and Irenimus stolidus Broun. Weevils were exposed to parasitoids and then examined 6, 10 and 15 days post-parasitism for parasitoid stage and size, and teratocyte number and size. In all hosts, teratocyte numbers decreased and size increased as parasitoid development progressed, although 6 days after parasitism, fewer, larger teratocytes were found in I. stolidus than S. discoideus or N. cervina. In weevils containing second-third instar parasitoid larvae, the most permissive hosts, S. discoideus and N. cervina contained more teratocytes than the least permissive hosts I. stolidus and R. conicus. Host gender influenced some aspects of parasitoid and teratocyte development. Total teratocyte volume was greater in female than male S. discoideus at all sampling times, and at 10 days post-parasitism in N. cervina. A possible relationship between host suitability and teratocyte development is discussed.     

Microplitis croceipes teratocytes cause developmental arrest of Heliothis virescens larvae

Microplitis croceipes teratocytes placed into nonparasitized Heliothis virescens larvae survived in the absence of a parasitoid larva and caused developmental changes in the host. Expressions of these changes included delayed larval mortality, incomplete larval-pupal ecdysis, or delayed pupation. Two day old 4th stadium H. virescens larvae were more sensitive to injected teratocytes than were 5th stadium larvae. Three day old teratocytes were more effective than were 6 day old teratocytes. The degree of response was related to the number of injected teratocytes. For example, 750 three day old teratocytes (the approximate number from a single parasitoid egg) caused delayed larval mortality in 96% of the treated larvae whereas 175 three day old teratocytes caused delayed larval mortality in only 33% of the treated larvae. Even a dose of 80 teratocytes resulted in 15% incomplete larval-pupal ecdysis compared to 0% for controls. Treatment with hemocyte-and teratocyte-free hemolymph from parasitized larvae, hemocytes from nonparasitized H. virescens, unfertilized M. croceipes eggs, Cotesia congregata teratocytes, or Micrococcus lysodeikticus cells all had very little effect either on larval growth or development time.     

Quantification and development of teratocytes in novel-association host-parasitoid combinations

We studied the development of teratocytes derived from two congeneric gregarious endoparasitic species, Cotesia chilonis and C. flavipes, parasitizing two congeneric novel hosts, Diatraea saccharalis and D. grandiosella. The host-parasitoid combinations studied allowed us to investigate relationships between host suitability and teratocyte development. D. saccharalis was a suitable host for both parasitoids, whereas D. grandiosella was suitable for C. chilonis development but often encapsulated C. flavipes progeny. Encapsulation of C. flavipes by D. grandiosella commenced around the time of parasitoid egg hatch, when teratocytes were released into the host's hemolymph. The gregarious parasitoids studied here released about 200 teratocytes per egg. Both absolute and normalized (teratocytes/parasitoid) numbers decreased over time. D. saccharalis supported more C. flavipes-derived teratocytes than D. grandiosella, possibly because of the unsuitability of the latter host. On intermediate assay days the number of C. flavipes-derived teratocytes was greater than for C. chilonis. However, C. chilonis-derived teratocytes grew larger than C. flavipes. Teratocytes in all host-parasitoid combinations doubled in size during parasitoid development. Teratocytes generally grew larger in D. grandiosella, which was a less suitable host.     

Host regulation by the aphid parasitoid Aphidius ervi: the role of teratocytes

The biochemical profile and metabolism of Acyrthosiphon pisum(Harris) (Homoptera, Aphididae) are markedly altered and redirected in response to parasitization by the endophagous braconid Aphidius erviHaliday (Hymenoptera, Braconidae). In the present study, the role played in the host regulation process by teratocytes, cells deriving from the dissociation of the embryonic membrane of the parasitoid, is taken into consideration. The protein synthesis activity of these cells of embryonic origin is analysed in vitroand an essential characterization of those proteins de novosynthesized and released in the incubation medium is provided. Teratocytes, obtained by dissecting parasitized host aphids, 3, 4 and 5 days after parasitoid oviposition, were incubated in vitroand, at the end of the incubation period, were separated from the medium for SDS-PAGE analysis of both cellular and secreted proteins. Various cellular proteins were more abundant as the time between parasitization and teratocyte collection increased. Furthermore, two proteins, showing an approximate molecular mass of 15 kD (p15) and 45 kD (p45) respectively, were abundantly secreted in the incubation medium by 5 day-old teratocytes. Incubations in presence of ³⁵S radiolabelled amino acids indicated that p15 and p45 are both synthesized by A. erviteratocytes. The amino acid composition of these two proteins was similar to that reported for other insect proteins with a demonstrated nutritional function. The p45 protein was found to be glycosylated. A tentative physiological model describing the host regulation role played by different parasitoid-derived factors is proposed.     

Expression and characterization of a novel teratocyte protein of the braconid,Microplitis croceipes (cresson)

Microplitis croceipes wasps overcome host immunity by inducing changes in host physiology using factors derived from the embryo and/or larva. Teratocytes of some parasitic wasps circulate in the host hemolymph after egg hatch and synthesize proteins (TSPs), some of which are secreted to alter host physiology in support of endoparasitoid development. TSPs appear to alter host physiology, at least in part, by inhibiting synthesis of certain proteins. M. croceipes teratocytes synthesize a 13.9 kDa protein (TSP14), which inhibits synthesis of host proteins that are linked to larval growth and development. A cDNA encoding TSP14 was generated by RT-PCR from teratocyte RNA, and cloned into yeast expression vectors to produce sufficient recombinant protein for functional analyses. RecTSP14 was produced using the yeast expression system at a concentration of more than 300 micrograms/L. The recTSP14 inhibited in vitro translation of larval Heliothis virescens RNA, with the activity sensitive to boiling, protein concentration, incubation time, and storage temperatures. Although recTSP14 inhibited translation of some cellular RNAs in vitro, the in vivo incorporation of [35S]-methionine into proteins of selected insect and mammalian cell lines was not inhibited. These findings suggest that recTSP14 entry is cell type-specific and required to inhibit synthesis of target protein(s).     

Growth patterns of Microplitis rufiventris (Hym., Braconidae) teratocytes in Spodoptera littoralis (Lep., Noctuidae) larvae treated with a chitin synthesis inhibitor

The influence of sublethal dietary levels (0.4 p.p.m.) of a chitin synthesis inhibitor lefenuron { N -[2,5-dichloro-4-(1,1,2,3,3,3-hexafluoropropoxy)-phenylaminocarbonyl]-2,6-difluorobenzamide} on the development of Microplitis rufiventris teratocytes was investigated. Observations and data were taken on maturing teratocytes, i.e. at the end of parasitoid development. When embryogenesis of the parasitoid eggs was initiated in treated hosts, the dissociated cells of the embryonic membrane of some of the parasitoid larvae did not liberate in the haemolymph of the host, whereas in the other hosts, the released ones as individual cells scored the largest mean number per host larva. When young teratocytes were exposed to the treatment they were found in some cases clumping in masses or clustered around the parasitoid larvae. In treated hosts, different size classes of teratocytes co-existed. Larger and morphological abnormal cells were more common in hosts which contained older teratocytes at the time of treatments. Abnormal teratocytes were associated with developmental abnormalities of parasitoid larvae, possibly due to deterioration of the host environment. This observation was common in treated hosts and in few cases of untreated ones. A significant decrease in teratocyte size occurred 24 h after parasitoid emergence. Observations and results obtained in the present work suggested that: (1) teratocyte may function in controlling the growth and successful development of parasitoid larvae; (2) they might have a protective function through sequestering abnormal materials issuing in the haemolymph of the host either during the course of parasitoid development or due to external stress; and (3) number of teratocyte cells liberated from the embryonic membrane of parasitoid egg is not constant.     

Ultrastructure of Microplitis croceipes (Cresson) (Braconidae : Hymenoptera) teratocytes

The developing embryo of the braconid, Microplitis croceipes (Braconidae : Hymenoptera), is encased in an extraembryonic serosal membrane. Hatching of the parasitoid within the larva of its habitual host, Hehothis virescens (Noctuidae : Lepidoptera), is initiated about 40 hr after oviposition when held at 25 ± 2°C. At this time, the monolayered serosal membrane begins to dissociate into individual cells (teratocytes). After dissociation, teratocytes become dispersed in the hemolymph of the host. The average number of teratocytes released from each parasitoid embryo is 914 ± 43. Teratocytes average 14.1 ± 2.4 μm in diameter when first released, and reach a maximum average diameter of 68.1 ± 4.6 μm 6 days after liberation. Newly released teratocytes have ovoid nuclei, simple mitochondria and a limited number of profiles of the endoplasmic reticulum, all of which indicate relative metabolic inactivity. The ramified nuclei, extensive endoplasmic reticulum, polymorphic mitochondria and accumulation of glycogen granules and lipid droplets observed in older teratocytes provide circumstantial evidence that protein synthesis is occurring. Within hours after dissociation, microvilli begin to cover the surface of the teratocytes. Anatomical deformation (blebs) that occurred on some older (8-day-old) teratocytes probably resulted from enlargement or expansion of microvilli.     

Effects of Microplitis croceipes Teratocytes on Host Haemolymph Protein Content and Fat Body Proliferation

Qualitative and quantitative changes in haemolymph proteins in Heliothis virescens were observed in larvae injected with either Microplitis croceipes teratocytes or teratocyte secreted proteins (TSP). Haemolymph protein titres in hosts receiving either 0.5 or 1 larval equivalent (LE) of teratocytes were similar to those of parasitized larvae, whereas a single injection of 4LE of TSP was required to induce a similar response. SDS-PAGE showed that the 82kDa monomer of riboflavin-binding protein and the 74/76kDa monomers of storage proteins were significantly reduced in parasitized larvae and in nonparasitized larvae treated with TSP. Concentrations of a 155kDa monomer (insectacyanin chromoprotein) also were reduced in parasitized larvae and those injected with either teratocytes or TSP. Two monomers (56 and 60kDa) were unique to parasitized larvae. Treated larvae required several days longer than controls to reach a comparable premetamorphic stage (burrowing-digging). Reductions in fat body proliferation similar to those seen in parasitized larvae were observed in larvae treated with either 1LE of teratocytes, or with 2 or 4LEs of TSP. Perivisceral fat body weights from larvae treated with either 0.25 or 0.5LE of teratocytes were significantly reduced, but less so than those which received 1LE. Thus, fat body proliferation in both teratocyte- and TSP-treated larvae was inhibited in a dose-dependent manner. Both light- and transmission electron microscopy observations revealed cytological differences in fat body tissues of larvae injected with either teratocytes or TSP from the condition observed in parasitized larvae and noninjected controls. Gross dissection of periviseral fat body from parasitized, teratocyte-injected and TSP-injected larvae showed tissue much less developed and differing considerably in appearance from controls. Observed differences included reduced size and/or number of lipid bodies and qualitative and quantitative changes in other cytoplasmic organelles.     

Juvenile hormone esterase activity and ecdysteroid titer in Heliothis virescens larvae injected with Microplitis croceipes teratocytes

Juvenile hormone esterase (JHE) activity in the hemolymph of 5th-instar Heliothis virescens larvae injected with Microplitis croceipes teratocytes was inversely related to the number of teratocytes injected. JHE activity in the hemolymph of larvae injected with 750 3-day-old teratocytes (the approximate number from one parasitoid embryo) was depressed to less than 5% of those levels found in control larvae. During the latter portion of the digging stage and in the burrowing-digging (BD) stage JHE activity in larvae treated with 350 teratocytes was approximately 40% of control values. However, injection of 180 teratocytes did not significantly affect JHE titers. Two-day-old teratocytes caused the greatest reduction in JHE titer with decreasing effects observed with injections of 3- to 6-day-old teratocytes. Nevertheless, because 2-day-old teratocytes were difficult to separate from host hemocytes, 3-day-old teratocytes were used in most of these studies. Injections of nonparasitized H. virescens hemolymph plasma, Micrococcus luteus bacterial cell walls, washed M. croceipes eggs, or teratocytes from Cotesia congregata did not depress JHE titers. Teratocyte injections also significantly reduced growth of host fat body. Ecdysteroid titers in cell formation, day 2 (CF2) larvae injected as new 5th instars with 350 3-day-old teratocytes failed to increase, as compared to noninjected and saline-injected controls. An injection of 1 μg/larva of 20-hydroxyecdysone at the BD stage permitted normal pupation in 50% of the teratocyte-treated larvae as compared to 0% pupation for teratocyte-treated control larvae not treated with 20-hydroxyecdysone. Teratocytes seem to be responsible for the inhibition of JHE release and thus indirectly impact on ecdysteroid titers. © 1992 Wiley-Liss, Inc.     

畸形细胞在协调寄生蜂同其寄主 相互关系中的作用

膜翅目内寄生的茧蜂科(Ｂｒａｃｏｎｉｄａｅ)、广腹细蜂科(Ｐｌａｔｙｇａｓｔｅｒｉｄａｅ)、缘腹卵蜂科(Ｓｃｅｌｉｏｎｉｄａｅ)中,蜂卵孵化时,起胚胎营养作用的浆膜(ｓｅｒｏｓａｌｍｅｍｂｒａｎｅ)释放到寄主血淋巴中,分散后游离形成畸形细胞。赤眼蜂科(Ｔｒｉｃｈ...     

畸形细胞在协调寄生蜂同其寄主 相互关系中的作用

Teratocytes originating from the serosal membrane of an endoparasitic wasp embryo, which is one of the species found in the Hymenopteran families Braconidae, Platygasteridae and Socelionidae, act an important role in coordinating the relationship between the larval wasp and its host. It may be concluded that inhibiting the immune system of the host, regulating development of the host and nourishing the parasitoid larva are three major functions of the teratocytes. By means of releasing inhibitor of phenol oxidase, a fundamental ingredient of the immune system of the host, the teratocytes prevent the host from encapsulation, which is fatal to the parasitoid larva. In the process of parasitism, the teratocytes reduce ecdysteroid titer and inhibit the metabolism of JH in the hemoymph of the host, and subsequently result in the developmental arrest of the host larva. On the other hand, the teratocytes secrete proteins or present themselves as food to the parasitoid. Although all those above are generally acknowledged functions of the teratocytes, there are other putative functions of the teratocytes, such as secreting histolytic enzymes and mycostatic material, and protecting the host from overharmed by the parasitoid before the parasitoid emerges from its host. The studies in this area can provide us more understanding and knowledge about the relationship between a parasitoid and its host.     

Teratocytes: Growth and numbers in the hemocoel of Heliothis virescens attacked by Microplitis croceipes

When the egg of Microplitis croceipes hatches in its host, Heliothis virescens, spherical cells (teratocytes) from the extraembryonic membrane are released into the host's hemolymph. Approximately 750 teratocytes are liberated from the parasitoid egg, and they average 10.5 μm in diameter when released. These cells increase in size, reaching a maximum average diameter of 140 μm in 8–9 days.The developing parasitoid emerges from the host in 9 days. The host remains alive and contains approximately 750 teratocytes, indicating that the teratocytes are not consumed by the parasitoid. These results suggest that teratocytes are not a direct source of nutrition for the developing parasitoid.Observations showed that encapsulation can occur in the presence of teratocytes even following the emergence of the parasitoid. The results indicate that teratocytes do not block the host's ability to encapsulate certain foreign materials within its hemocoel.     

Larvae of an endoparasitoid,Cotesia kariyai (Hymenoptera: Braconidae), feed on the host fat body directly in the second stadium with the help of teratocytes

Larvae of a gregarious endoparasitoid, Cotesia kariyai (Watanabe), grew rapidly during the second stadium in the host. The fat body of a Pseudaletia host parasitized by C. kariyai was completely consumed by 10 d, just before larval emergence. It seemed hard to explain the growth of the second instar parasitoids and the rapid consumption of the fat body only by ingestion of hemolymph converted from the fat body or other organs of the host. Paraffin sections of the parasitized host revealed that many teratocytes were attached to the surface of the fat body in many sites and destroyed the fat body tissue locally. Zymography of proteins released from the teratocytes revealed that the teratocytes 4 to 9 days after parasitization showed collagenase activity (as a gelatinase). Further, 1st instar parasitoids which were transplanted together with teratocytes into unparasitized hosts preconditioned with C. kariyai polydnavirus (CkPDV) plus venom, grew normally to the 2nd stadium. Abnormal growth of parasitoid larvae was observed when parasitoid larvae were transplanted without teratocytes. These results suggest that the teratocytes attach to the outer sheath of the fat body, secrete an enzyme that makes a hole in the matrix of the fat body, thus allowing the second instar parasitoid to ingest the content of the fat body.     

Systematic analysis of a wasp parasitism arsenal

Parasitoid wasps are among the most diverse insects on earth with many species causing major mortality in host populations. Parasitoids introduce a variety of factors into hosts to promote parasitism, including symbiotic viruses, venom, teratocytes and wasp larvae. Polydnavirus‐carrying wasps use viruses to globally suppress host immunity and prevent rejection of developing parasites. Although prior results provide detailed insights into the genes viruses deliver to hosts, little is known about other products. RNAseq and proteomics were used to characterize the proteins secreted by venom glands, teratocytes and larvae from Microplitis demolitor, which carries M. demolitor bracovirus (MdBV). These data revealed that venom glands and teratocytes secrete large amounts of a small number of products relative to ovaries and larvae. Venom and teratocyte products exhibited almost no overlap with one another or MdBV genes, which suggested that M. demolitor effector molecules are functionally partitioned according to their source. This finding was well illustrated in the case of MdBV and teratocytes. Many viral proteins have immunosuppressive functions that include disruption of antimicrobial peptide production, yet this study showed that teratocytes express high levels of the antimicrobial peptide hymenoptaecin, which likely compensates for MdBV‐mediated immunosuppression. A second key finding was the prevalence of duplications among genes encoding venom and teratocyte molecules. Several of these gene families share similarities with proteins from other species, while also showing specificity of expression in venom glands or teratocytes. Overall, these results provide the first comprehensive analysis of the proteins a polydnavirus‐carrying wasp introduces into its host.