Lepidopteran Hosts Research Articles

Potential Host Range of the Larval Endoparasitoid Cotesia vestalis (=plutellae) (Hymenoptera: Braconidae).

Many parasitoid wasps are highly specialized in nature, attacking only one or a few species of hosts. Host range is often determined by a range of biological and ecological characteristics of the host including diet, growth potential, immunity, and phylogeny. The solitary koinobiont endoparasitoid wasp, Cotesia vestalis, mainly parasitizes diamondback moth (DBM) larvae in the field, although it has been reported that to possess a relatively wide lepidopteran host range. To better understand the biology of C vestalis as a potential biological control of hosts other than the DBM, it is necessary to determine suitability for potential hosts. In this study, the potential host range of the wasp and its developmental capacity in each host larva were examined under laboratory conditions using 27 lepidopteran species from 10 families. The wasp was able to parasitize 15 of the 27 species successfully. Some host species were not able to exclude C vestalis via their internal physiological defenses. When parasitization was unsuccessful, most hosts killed the parasitoid at the egg stage or early first-instar stage using encapsulation, but some host species disturbed the development of the parasitoid at various stages. No phylogenetic relationships were found among suitable and unsuitable hosts, revealing that host range in some endoparasitoids is not constrained by relatedness among hosts based on immunity.

High temperature induces downregulation of polydnavirus gene transcription in lepidopteran host and enhances accumulation of host immunity gene transcripts

Endoparasitoids face the challenge of overcoming the immune reaction of their hosts, which typically consists of encapsulation and melanisation of parasitoid eggs or larvae. Some endoparasitic wasps such as the solitary Tranosema rostrale (Hymenoptera: Ichneumonidae) that lay their eggs in larvae of the spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae), have evolved a symbiotic relationship with a polydnavirus (PDV), which in turn helps them suppress the host’s immune response. We observed an increase in mortality of immature T. rostrale with increasing temperature, and we tested two hypotheses about the mechanisms involved: high temperatures (1) hamper the expression of T. rostrale PDV genes and (2) enhance the expression of spruce budworm immunity-related genes. Dissections of parasitized spruce budworm larvae reared at 30°C revealed that most parasitoid eggs or larvae had died as a result of encapsulation and melanisation by the host. A qPCR analysis of T. rostrale PDV (TrIV) gene expression showed that the transcription of several TrIV genes in host larvae was downregulated at high temperature. On the other hand, encapsulation, but not melanisation, of foreign bodies in spruce budworm larvae was enhanced at high temperatures, as shown by the injection of Sephadex™ beads into larvae. However, at the molecular level, the transcription of genes related to spruce budworm’s melanisation process (prophenoloxidase 1 and 2) was upregulated. Our results support the hypothesis that a temperature-dependent increase of encapsulation response is due to the combined effects of reduced expression of TrIV genes and enhanced expression of host immune genes.

Transcript Abundance of Photorhabdus Insect-Related (Pir) Toxin in Manduca sexta and Galleria mellonella Infections.

In this study, we assessed pirAB toxin transcription in Photorhabdus luminescens laumondii (strain TT01) (Enterobacteriaceae) by comparing mRNA abundance under in vivo and in vitro conditions. In vivo assays considered both natural and forced infections with two lepidopteran hosts: Galleria mellonella and Manduca sexta. Three portals of entry were utilized for the forced infection assays: (a) integument; (b) the digestive route (via mouth and anus); and (c) the tracheal route (via spiracles). We also assessed plu4093-2 transcription during the course of a natural infection; this is when the bacteria are delivered by Heterorhabditis bacteriophora nematodes. Transcript abundance in G. mellonella was higher than in M. sexta at two of the observed time points: 15 and 18 h. Expression of pirAB plu4093-2 reached above endogenous control levels at 22 h in G. mellonella but not in M. sexta. Overall, pirAB plu4093-2 transcripts were not as highly expressed in M. sexta as in G. mellonella, from 15 to 22 h. This is the first study to directly compare pirAB plu4093-2 toxin transcript production considering different portals of entry.

Midgut microbiota and host immunocompetence underlie Bacillus thuringiensis killing mechanism

Bacillus thuringiensis is a widely used bacterial entomopathogen producing insecticidal toxins, some of which are expressed in insect-resistant transgenic crops. Surprisingly, the killing mechanism of B. thuringiensis remains controversial. In particular, the importance of the septicemia induced by the host midgut microbiota is still debated as a result of the lack of experimental evidence obtained without drastic manipulation of the midgut and its content. Here this key issue is addressed by RNAi-mediated silencing of an immune gene in a lepidopteran host Spodoptera littoralis, leaving the midgut microbiota unaltered. The resulting cellular immunosuppression was characterized by a reduced nodulation response, which was associated with a significant enhancement of host larvae mortality triggered by B. thuringiensis and a Cry toxin. This was determined by an uncontrolled proliferation of midgut bacteria, after entering the body cavity through toxin-induced epithelial lesions. Consequently, the hemolymphatic microbiota dramatically changed upon treatment with Cry1Ca toxin, showing a remarkable predominance of Serratia and Clostridium species, which switched from asymptomatic gut symbionts to hemocoelic pathogens. These experimental results demonstrate the important contribution of host enteric flora in B. thuringiensis-killing activity and provide a sound foundation for developing new insect control strategies aimed at enhancing the impact of biocontrol agents by reducing the immunocompetence of the host.

Permissiveness of lepidopteran hosts is linked to differential expression of bracovirus genes

Polydnaviruses in the genus Bracovirus (BV) are associated with parasitoid wasps in the family Braconidae. BV-carrying wasps rely on their associated viruses to parasitize permissive hosts but also occasionally oviposit into host species that are non-permissive. Here, we studied Microplitis demolitor and M. demolitor bracovirus (MdBV) in Chrysodeixis includens, a permissive host, and Trichoplusia ni, which is usually non-permissive. M. demolitor laid eggs and injected MdBV into both hosts but almost no wasp offspring developed in T. ni. MdBV DNA similarly persisted in both host species, but deep sequencing data showed that transcript abundance for most viral genes was higher in C. includens than T. ni. Overall, our results identify lower expression of MdBV genes as an important factor in the non-permissiveness of T. ni. However, certain genes with functions in immunosuppression were sufficiently expressed to have similar effect in T. ni and C. includens.

Coleopteran and Lepidopteran Hosts of the Entomopathogenic Genus Cordyceps sensu lato

Entomophthoralean and ascomycetous fungi are the two major groups known to parasitize arthropods in almost every terrestrial habitat of the earth. Within Ascomycota, Cordyceps sensu lato is a large genus with more than 400 spp. described on numerous orders of Arthropoda. Among the hosts of Cordyceps, Coleoptera and Lepidoptera are the two major orders. Out of the estimated 200 Cordyceps spp. recorded on coleopteran and lepidopteran hosts, we have documented 92 spp. based on the available information of their host species. Among coleopteran hosts, Scarabaeidae and Elateridae are the two major families. Similarly, among lepidopterans, Hepialidae is the largest host family. Cordyceps militaris shows the widest host range, extending to 2 orders, 13 families, and 32 spp. We hope such accumulative work will be useful as a quick reference for interested biologists, forest ecologists, biocontrol researchers, and fungal and insect taxonomists to apprehend host range and host specificities of Cordyceps fungi.

Parasitism and Suitability of Fertilized and Nonfertilized Eggs of the Rice Striped Stem Borer, Chilo suppressalis (Lepidoptera: Crambidae), for Trichogramma Parasitoids.

Trichogramma spp. are egg parasitoids largely used worldwide for biological control of lepidopteran pests. Host quality (related to host size and age, host species, etc.) can influence parasitism preference and fitness of the parasitoid offspring. However, few studies have documented the performance of Trichogramma parasitoids when they parasitize nonfertilized eggs of their lepidopteran hosts. In the present study, we investigated the performance of three indigenous Trichogramma species ( Trichogramma japonicum Ashmead, Trichogramma dendrolimi Matsumura, and Trichogramma chilonis Ishii) on fertilized and nonfertilized eggs of their native host, Chilo suppressalis (Walker). The results showed that the three Trichogramma species tested could recognize fertilized and nonfertilized eggs of C. suppressalis . The different fertilization status eggs of C. suppressalis were all accepted by T. japonicum , T. chilonis , and T. dendrolimi with a clear tendency that they preferred to parasitize fertilized eggs whether under no-choice or choice conditions. All Trichogramma species successfully completed development in parasitized eggs of C. suppressalis regardless if the host eggs were fertilized or not. There was similar adult emergence rate, development time, and female progeny between fertilized and nonfertilized eggs for all Trichogramma species with an exception of development for T. chilonis . Trichogramma chilonis had shorter developmental time in fertilized eggs than in nonfertilized ones. Whether in fertilized or nonfertilized eggs, T. dendrolimi had longer development time than other two Trichogramma species. However, T. dendrolimi had the largest percentage of female progeny on fertilized eggs, followed by T. japonicum , and T. chilonis had the least percentage of female progeny. The present study indicates that Trichogramma parasitoids (i) can distinguish fertilized vs. nonfertilized host eggs, and (ii) could use nonfertilized host eggs for successful offspring development (despite showing lower preference for such eggs) [corrected].

Pathogenicity, morphology, and characterization of a Nosema fumiferanae isolate (Microsporidia: Nosematidae) from the light brown apple moth, Epiphyas postvittana (Lepidoptera: Tortricidae) in California

We recently discovered infections by a microsporidium closely related to Nosema fumiferanae in field populations of the light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), in the San Francisco region of California. E. postvittana originates from Australia and was first detected in California in 2006; therefore, our aim was to identify and determine the origin of the Nosema isolate. We characterized the pathogenicity, transmission pathways, and ultrastructure of this new Nosema isolate. In addition, we sequenced fragments of commonly used genetic markers (ITS, SSU, and RPB1), and examined the phylogenetic relationships between the Nosema isolate and other microsporidian species commonly found in lepidopteran hosts. The pathogenicity of the Nosema isolate was investigated by infecting second instar larvae of E. postvittana. Larval and pupal survivorship were reduced by 7% and 13% respectively, and pupation occurred 1–2d later in infected individuals than in healthy individuals. Emerging infected females died 5d earlier than healthy females, and daily fecundity was 22% lower. Hatch rate also was 22% lower for eggs oviposited by infected females. Vertical transmission was confirmed; spores were present in 68% of egg masses and 100% of the surviving larvae from infected females. Ultrastructure images, together with sequences from selected genetic markers, confirmed the Nosema isolate to be a member of the Nosema fumiferanae species complex (Nosema fumiferanae postvittana subsp. n.). The association of this pathogen with E. postvittana contributes further to the biotic resistance that E. postvittana has experienced since its introduction to California.

Discovery of the firstAximopsis(Hymenoptera: Eurytomidae) Parasitoid of Lepidoptera in Brazil and Notes on Its Biology

Eucalyptus cloeziana F. Muell. (Myrtales: Myrtaceae) and the tropical almond Terminalia catappa L. (Myrtales: Combretaceae) are widely cultivated in urban and forest areas of many countries where biological control is the most-preferred method to control insects. Aximopsis sp. (Hymenoptera: Eurytomidae) is reported for the first time in Brazil in a new group of lepidopteran hosts. Individuals of this species emerged from the pupae of Thyrinteina arnobia arnobia Stoll (Lepidoptera: Geometridae) and Thagona tibialis Walker (Lepidoptera: Lymantriidae) that developed from larvae defoliating E. cloeziana and T. catappa plants on the campus of the Universidade Federal de Vicosa (UFV) in Vicosa, Minas Gerais, Brazil. Aximopsis sp. was identified by comparing it with species of this group as described for the Neotropical region. Voucher specimens were deposited in the Centre de Cooperation Internationale en Recherche Agronomique pour le Developpement (CIRAD), France. Twenty new pupae each of Anticarsia gemmatalis Hubner (Lepidoptera: Noctuidae) and Tenebrio molitor L. (Coleoptera: Tenebrionidae) were held individually in test tubes with a drop of honey as food and 3 mated Aximopsis sp. females for 2 d. Aximopsis sp. parasitized 20% of the T. molitor pupae but none of the A. gemmatalis pupae. The duration of the life cycle (egg to adult), parasitism and emergence rates, total individuals emerged from each pupa, sex ratio, length of the body and width of the head capsule, and the longevities of Aximopsis sp. males and females that emerged from parasitized T. molitor pupae were evaluated. The duration of the life cycle of Aximopsis sp. was 14 ± 2 d. An average of 62 ± 5 Aximopsis sp. individuals emerged from each T. molitor pupa, and their sex ratio was 0.96 ± 0.02. The total number of parasitoids that emerged was 248 individuals. Measurements of characters of progeny Aximopsis sp. females (n = 10) and males (n = 5), respectively, were as follows: body length: 3.50 mm (2.40–3.80 mm) and 1.99 mm (1.97–2.02 mm); head capsule width: 0.63 mm (0.58–0.72 mm) and 0.48 mm (0.46–0.51 mm), and longevity: 6 ± 1 d and 4 ± 1 d. These results open prospects for investigations of biological control of pests with this natural enemy.

Gasmin (BV2-5), a polydnaviral-acquired gene in Spodoptera exigua. Trade-off in the defense against bacterial and viral infections

Thousands of Hymenopteran endoparasitoids have developed a unique symbiotic relationship with viruses named polydnavirus (PDVs). These viruses immunocompromise the lepidopteran host allowing the survival of the wasp eggs. In a previous work, we have shown the horizontal transfer of some polydnaviral genes into the genome of the Lepidoptera, Spodoptera exigua. One of these genes, BV2-5 (named gasmin) interferes with actin polymerization, negatively affecting the multiplication of baculovirus in cell culture. In this work, we have focused in the study of the effect of Gasmin expression on different aspects of the baculovirus production. In addition, and since actin polymerization is crucial for phagocytosis, we have studied the effect of Gasmin expression on the larval interaction with bacterial pathogens. Over-expression of Gasmin on hemocytes significantly reduces their capacity to phagocytize the pathogenic bacteria Bacillus thuringiensis. According to these results, gasmin domestication negatively affects baculovirus replication, but increases larvae susceptibility to bacterial infections as pay off. Although the effect of Gasmin on the insect interaction with other pathogens or parasitoids remain unknown, the opposite effects described here could shape the biological history of this species based on the abundance of certain type of pathogens as suggested by the presence of truncated forms of this protein in several regions of the world.

Recurrent Domestication by Lepidoptera of Genes from Their Parasites Mediated by Bracoviruses

Bracoviruses are symbiotic viruses associated with tens of thousands of species of parasitic wasps that develop within the body of lepidopteran hosts and that collectively parasitize caterpillars of virtually every lepidopteran species. Viral particles are produced in the wasp ovaries and injected into host larvae with the wasp eggs. Once in the host body, the viral DNA circles enclosed in the particles integrate into lepidopteran host cell DNA. Here we show that bracovirus DNA sequences have been inserted repeatedly into lepidopteran genomes, indicating this viral DNA can also enter germline cells. The original mode of Horizontal Gene Transfer (HGT) unveiled here is based on the integrative properties of an endogenous virus that has evolved as a gene transfer agent within parasitic wasp genomes for ≈100 million years. Among the bracovirus genes thus transferred, a phylogenetic analysis indicated that those encoding C-type-lectins most likely originated from the wasp gene set, showing that a bracovirus-mediated gene flux exists between the 2 insect orders Hymenoptera and Lepidoptera. Furthermore, the acquisition of bracovirus sequences that can be expressed by Lepidoptera has resulted in the domestication of several genes that could result in adaptive advantages for the host. Indeed, functional analyses suggest that two of the acquired genes could have a protective role against a common pathogen in the field, baculovirus. From these results, we hypothesize that bracovirus-mediated HGT has played an important role in the evolutionary arms race between Lepidoptera and their pathogens.

Development of a Real-Time qPCR Assay for Quantification of Covert Baculovirus Infections in a Major African Crop Pest.

Many pathogens and parasites are present in host individuals and populations without any obvious signs of disease. This is particularly true for baculoviruses infecting lepidopteran hosts, where studies have shown that covert persistent viral infections are almost ubiquitous in many species. To date, the infection intensity of covert viruses has rarely been quantified. In this study, we investigated the dynamics of a covert baculovirus infection within the lepidopteran crop pest Spodoptera exempta. A real-time quantitative polymerase chain reaction (qPCR) procedure using a 5' nuclease hydrolysis (TaqMan) probe was developed for specific detection and quantification of Spodoptera exempta nucleopolyhedrovirus (SpexNPV). The qPCR assay indicated that covert baculovirus dynamics varied considerably over the course of the host life-cycle, with infection load peaking in early larval instars and being lowest in adults and final-instar larvae. Adult dissections indicated that, contrary to expectation, viral load aggregation was highest in the head, wings and legs, and lowest in the thorax and abdomen. The data presented here have broad implications relating to our understanding of transmission patterns of baculoviruses and the role of covert infections in host-pathogen dynamics.

Digital gene expression analysis of Helicoverpa armigera in the early stage of infection with Helicoverpa armigera nucleopolyhedrovirus

Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) is an obligatory and lethal parasite of the cotton bollworm and has been extensively used in China for the control of this notorious pest. Digital gene expression (DGE) analysis was adopted for an overall comparison of transcriptome profiling between HearNPV-infected and control healthy Helicoverpa armigera larvae during an early stage post-inoculation. A total of 908 differentially expressed genes (DEGs) were identified, of which 136 were up-regulated and 597 were down-regulated. GO category and KEGG pathway analysis demonstrated that the identified DEGs involved in ribosome biogenesis, aminoacyl-tRNA biosynthesis, protein processing in endoplasmic reticulum, biosynthesis of valine, leucine, isoleucine and the spliceosome were significantly down-regulated, whereas genes involved in pancreatic secretion, protein digestion and absorption and salivary secretion showed obviously up-regulated transcription. The DEGs were verified by quantitative real-time PCR, and genes that participated in defensive response, nutritional digestion and developmental regulation exhibited specific expression patterns in a continuous time-course assessment. These results provide basic data for future research on the molecular mechanism of HearNPV infection and the interactions between lepidopteran hosts and their specific NPV parasites.

Field Host Range ofApanteles opuntiarum(Hymenoptera: Braconidae) in Argentina, a Potential Biocontrol Agent ofCactoblastis cactorum(Lepidoptera: Pyralidae) in North America

Summary Field exploration in Argentina for cactophagous lepidopteran hosts parasitized by the recently described braconid parasitoid Apanteles opuntiarum Martinez & Berta (Hymenoptera: Braconidae) revealed a host range restricted to Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) and C. doddi. Field collections of cactophagous larvae parasitized by the congeneric and sympatric parasitoid A. alexanderi included the host species C. bucyrus, Sigelgaita nr. chilensis, Tucumania sp., Tucumania tapiacola and Salambona sp. If a narrow host range for A. opuntiarum is confirmed in quarantine with North American cactus-feeding species, then this parasitoid could be released as a biological control agent for C. cactorum with little or no risk to non-target species.

Attraction of egg-killing parasitoids toward induced plant volatiles in a multi-herbivore context

In response to insect herbivory, plants emit volatile organic compounds which may act as indirect plant defenses by attracting natural enemies of the attacking herbivore. In nature, plants are often attacked by multiple herbivores, but the majority of studies which have investigated indirect plant defenses to date have focused on the recruitment of different parasitoid species in a single-herbivore context. Here, we report our investigation on the attraction of egg parasitoids of lepidopteran hosts (Trichogramma brassicae and T. evanescens) toward plant volatiles induced by different insect herbivores in olfactometer bioassays. We used a system consisting of a native crucifer, Brassica nigra, two naturally associated herbivores [the butterfly Pieris brassicae (eggs and caterpillars) and the aphid Brevicoryne brassicae] and an alien invasive herbivore (eggs and caterpillars of the moth Spodoptera exigua). We found that Trichogramma wasps were attracted by volatiles induced in the plants by P. brassicae eggs, but not by those induced in the plants by S. exigua eggs, indicating the specificity of the plant responses toward lepidopteran herbivores. The results of the chemical analysis revealed significant differences between the volatile blends emitted by plants in response to attack by P. brassicae and S. exigua eggs which were in agreement with the behavioural observations. We investigated the attraction of Trichogramma wasps toward P. brassicae egg-induced volatiles in plants simultaneously attacked by larvae and nymphs of different non-hosts. The two chewing caterpillars P. brassicae and S. exigua, but not the phloem-feeding aphid B. brassicae, can disrupt the attraction of Trichogramma species toward P. brassicae egg-induced volatiles. Indirect plant defenses are discussed in the context of multiple herbivory by evaluating the importance of origin, dietary specialization and feeding guild of different attackers on the recruitment of egg-killing parasitoids.

Transcriptome analysis of the brain of the silkworm Bombyx mori infected with Bombyx mori nucleopolyhedrovirus: A new insight into the molecular mechanism of enhanced locomotor activity induced by viral infection

Baculoviruses have been known to induce hyperactive behavior in their lepidopteran hosts for over a century. As a typical lepidopteran insect, the silkworm Bombyx mori displays enhanced locomotor activity (ELA) following infection with B. mori nucleopolyhedrovirus (BmNPV). Some investigations have focused on the molecular mechanisms underlying this abnormal hyperactive wandering behavior due to the virus; however, there are currently no reports about B. mori. Based on previous studies that have revealed that behavior is controlled by the central nervous system, the transcriptome profiles of the brains of BmNPV-infected and non-infected silkworm larvae were analyzed with the RNA-Seq technique to reveal the changes in the BmNPV-infected brain on the transcriptional level and to provide new clues regarding the molecular mechanisms that underlies BmNPV-induced ELA. Compared with the controls, a total of 742 differentially expressed genes (DEGs), including 218 up-regulated and 524 down-regulated candidates, were identified, of which 499, 117 and 144 DEGs could be classified into GO categories, KEGG pathways and COG annotations by GO, KEGG and COG analyses, respectively. We focused our attention on the DEGs that are involved in circadian rhythms, synaptic transmission and the serotonin receptor signaling pathway of B. mori. Our analyses suggested that these genes were related to the locomotor activity of B. mori via their essential roles in the regulations of a variety of behaviors and the down-regulation of their expressions following BmNPV infection. These results provide new insight into the molecular mechanisms of BmNPV-induced ELA.

Scale-dependent, contrasting effects of habitat fragmentation on host-natural enemy trophic interactions

Habitat fragmentation can have contrasting effects on species and their interactions within communities, changing community structure and function. Parasitoids and pathogens are key natural enemies in invertebrate communities, but their responses to fragmentation have not been explored within the same community. This study aimed to explore the scale-dependent effects of habitat fragmentation on the population density of a lepidopteran host and particularly its trophic interactions with a specialist parasitoid and virus. Host density and host larval-mortality from the parasitoid and the virus were measured in twelve isolated sites and thirteen connected sites. An index of habitat isolation was created based on the amount of suitable habitat surrounding sites at a range of spatial scales (0.1–5 km radii), and the direct and indirect effects of habitat isolation were analysed using generalised linear mixed effects models. Consistent with predictions, habitat isolation had direct negative effects on host density at the smallest and largest spatial scales, and indirect negative effects on host mortality from the virus at the largest scale, but in contrast to predictions it had direct positive effects on parasitism at small and medium scales. Higher trophic level species may still display responses to habitat fragmentation contrary to predictions based on well supported theory and empirical evidence. The mechanisms underlying these responses may be elucidated by studying responses, contrary to expectations, shared by related species. Developing general predictions about the responses of host-pathogen interactions to fragmentation will require greater understanding of the system-specific mechanisms by which fragmentation can influence pathogen transmission.

Parasitism and survival rate of Diadegma fenestrale (Hymenoptera: Ichneumonidae) and DfIV gene expression patterns in two lepidopteran hosts

The genus Diadegma is a well-known parasitoid group and some are known to have symbiotic virus, polydnavirus (PDV). A novel IV was discovered from the calyx of Diadegma fenestrale female and sequenced its genome. D. fenestrale has more than two hosts, including potato tuber moth (PTM) and diamondback moth (DBM). D. fenestrale preferred PTM to DBM as hosts based on the oviposition and survival rate. Nevertheless, the developmental period and morphology of D. fenestrale were not significantly different between PTM and DBM. We compared DfIV gene expression patterns between PTM and DBM under various conditions to understand the phenomena. DfIV genes were more widely expressed in PTM with large numbers than in DBM after parasitized by D. fenestrale, particularly at the initial point. They showed differential expression patterns between two lepidopteran hosts. This DfIV gene expression plasticity showed a dependency on the lepidopteran host species and parasitization time, suggesting that it may contribute to increase the parasitoid survival rate. This might be one of the key elements that determine the symbiotic relationship between PDV and parasitoid.

Phylogeny of the parasitic wasp subfamily Euphorinae (Braconidae) and evolution of its host preferences

AbstractThe braconid subfamily Euphorinae is a large, cosmopolitan group of endoparasitoid wasps. The majority of species attack adult hosts, a strategy that is rare among parasitic wasps, but there are also many species that attack nymphs and larval stages. Euphorine hosts may belong to a variety of insect orders (Coleoptera, Hemiptera, Hymenoptera, Neuroptera, Psocoptera, Orthoptera and Lepidoptera) although most euphorine tribes are confined to Coleoptera. Here we investigate the phylogenetic relationships of the Euphorinae based on molecular data (3 kb of nucleotide data from four markers:18S,28S,CADandCOI) and propose a higher‐level classification based upon the resulting phylogeny. We also infer the evolution of host associations and discuss the diversification of the Euphorinae. Results from both Bayesian inference and maximum‐likelihood analysis show that the subfamily, as previously circumscribed, is paraphyletic. We propose that the subfamily be expanded to include the tribes Meteorini and Planitorini (Mannokeraia + Planitorus), so that it corresponds to a clade that is strongly supported as monophyletic in our analyses. Based on our results, a revised higher classification of the Euphorinae is proposed, in which 52 extant genera and 14 tribes are recognized. We reinstate the genusMicroctonusbelonging to the tribe Perilitini, and synonymizeUssurarideluswithHoldawayella, SinuatophoruswithEucosmophorus. Furthermore, we propose the following tribal rearrangements:SpathicopisandStenothremmaare transferred to Perilitini;Tuberidelus, EucosmophorusandPlynopsto Cosmophorini;Eccliturato Dinocampini;Chrysopophthorus, HoldawayellaandWesmaeliato Helorimorphini;ProclithroporusandHeiato Townesilitini. The monotypic tribe Cryptoxilonini is synonymized with Cosmophorini. The generaPygostolusandLitostolusare placed in a separate tribe, Pygostolini, previously recognized as a subtribe among the Centistini. Parsimony‐based ancestral state reconstructions suggest that the ancestor of Euphorinae was a parasitoid of lepidopteran larvae, and that a host shift to larval Coleoptera occurred only in one clade of the Meteorini, some members of which secondarily shifted back to larval lepidopteran hosts. In the remainder of the subfamily, there was an initial shift from larval to adult coleopterans, followed by subsequent shifts to adults or larvae of Hemiptera, Hymenoptera, Neuroptera, Orthoptera and Psocoptera.

Expression, delivery and function of insecticidal proteins expressed by recombinant baculoviruses.

Since the development of methods for inserting and expressing genes in baculoviruses, a line of research has focused on developing recombinant baculoviruses that express insecticidal peptides and proteins. These recombinant viruses have been engineered with the goal of improving their pesticidal potential by shortening the time required for infection to kill or incapacitate insect pests and reducing the quantity of crop damage as a consequence. A wide variety of neurotoxic peptides, proteins that regulate insect physiology, degradative enzymes, and other potentially insecticidal proteins have been evaluated for their capacity to reduce the survival time of baculovirus-infected lepidopteran host larvae. Researchers have investigated the factors involved in the efficient expression and delivery of baculovirus-encoded insecticidal peptides and proteins, with much effort dedicated to identifying ideal promoters for driving transcription and signal peptides that mediate secretion of the expressed target protein. Other factors, particularly translational efficiency of transcripts derived from recombinant insecticidal genes and post-translational folding and processing of insecticidal proteins, remain relatively unexplored. The discovery of RNA interference as a gene-specific regulation mechanism offers a new approach for improvement of baculovirus biopesticidal efficacy through genetic modification.