Jenny S. Cory

Ancient Coevolution of Baculoviruses and Their Insect Hosts

ABSTRACT If the relationships between baculoviruses and their insect hosts are subject to coevolution, this should lead to long-term evolutionary effects such as the specialization of these pathogens for their hosts. To test this hypothesis, a phylogeny of the Baculoviridae, including 39 viruses from hosts of the orders Lepidoptera, Diptera, and Hymenoptera, was reconstructed based on sequences from the genes lef-8 and ac22. The tree showed a clear division of the baculoviruses according to the order of their hosts. This division highlighted the need to reconsider the classification of the baculoviruses to include one or possibly two new genera. Furthermore, the specialization of distinct virus lineages to particular insect orders suggests ancient coevolutionary interactions between baculoviruses and their hosts.

Host ecology determines the relative fitness of virus genotypes in mixed-genotype nucleopolyhedrovirus infections

Mixed‐genotype infections are common in many natural host–parasite interactions. Classical kin‐selection models predict that single‐genotype infections can exploit host resources prudently to maximize fitness, but that selection favours rapid exploitation when co‐infecting genotypes share limited host resources. However, theory has outpaced evidence: we require empirical studies of pathogen genotypes that naturally co‐infect hosts. Do genotypes actually compete within hosts? Can host ecology affect the outcome of co‐infection? We posed both questions by comparing traits of infections in which two baculovirus genotypes were fed to hosts alongside inocula of the same or a different genotype. The host, Panolis flammea, is a herbivore of Pinus sylvestris and Pi. contorta. The pathogen, PfNPV (a nucleopolyhedrovirus), occurs naturally as mixtures of genotypes that differ, when isolated, in pathogenicity, speed of kill and yield. Single‐genotype infection traits failed to predict the ‘winning’ genotypes in co‐infections. Co‐infections infected and caused lethal disease in more hosts, and produced high yields, relative to single‐genotype infections. The need to share with nonkin did not cause fitness costs to either genotype. In fact, in hosts feeding on Pi. sylvestris, one genotype gained increased yields in mixed‐genotype infections. These results are discussed in relation to theory surrounding adaptive responses to competition with nonkin for limited resources.

Responses of Mamestra brassicae (Lepidoptera: Noctuidae) to crowding: interactions with disease resistance, colour phase and growth

This study examines phenotypic plasticity in relation to rearing density in larvae of the moth, Mamestra brassicae. Larval phase, growth rate, weight at moulting and susceptibility to disease were quantified when reared at five densities. Larvae develop more quickly, but attain a smaller size and are more susceptible to disease, when reared at high than at intermediate densities. They also exhibit a higher degree of melanisation than larvae reared at intermediate densities, or singly. A review of the literature suggests that a switch to a rapidly developing dark phase at high densities is a widespread phenomenon within the Lepidoptera. Rapid development at the expense of attaining a large size, and increased melanisation, are interpreted as adaptive responses to reach pupation before food supplies are depleted, as is likely when larval density is high. High susceptibility to viral infection at high density may be a result of physiological stress associated with rapid development, or due to a shift in allocation of resources from resistance to development: larvae that developed quickly were more susceptible to infection. Larvae reared singly appeared to be less fit than larvae reared at intermediate densities: they exhibited many of the characteristics of larvae reared at high density, particularly low weight, a right-hand skew in their weight frequency distribution, and high susceptibility to disease. I hypothesise that expression of resistance may be phenotypically plastic with regard to environment. Contact with other larvae may, up to a point, stimulate both growth and resistance to infection, for the risk of infection will increase with the density of conspecifics.

Fungal entomopathogens in a tritrophic context.

Variation in plant quality has an important impact on insect growth and development and there is considerable evidence that plants can also influence an insect’s natural enemies. Here we discuss the potential for plant-mediated effects on fungal entomopathogens. Fungi differ from other insect pathogens in that they infect an insect directly through its cuticle. This means that they are particularly vulnerable to changes in microclimate and properties of the insect cuticle. Potential direct and indirect mechanisms for plant-mediated effects on fungal entomopathogens are discussed. It is clear from these studies that fungal entomopathogens could be affected by plant volatiles and plant surface chemistry. Plant secondary chemicals can also inhibit fungal growth, potentially protecting the insect herbivore. However, the site of action and the mechanism behind these effects in plant-based studies is not always clear. The implications for biocontrol using fungal entomopathogens are discussed.

An experimental test of the independent action hypothesis in virus-insect pathosystems.

The ‘independent action hypothesis’ (IAH) states that each pathogen individual has a non-zero probability of causing host death and that pathogen individuals act independently. IAH has not been rigorously tested. In this paper, we (i) develop a probabilistic framework for testing IAH and (ii) demonstrate that, in two out of the six virus–insect pathosystems tested, IAH is supported by the data. We first show that IAH inextricably links host survivorship to the number of infecting pathogen individuals, and develop a model to predict the frequency of single- and dual-genotype infections when a host is challenged with a mixture of two genotypes. Model predictions were tested using genetically marked, near-identical baculovirus genotypes, and insect larvae from three host species differing in susceptibility. Observations in early-instar larvae of two susceptible host species support IAH, but observations in late-instar larvae of susceptible host species and larvae of a less susceptible host species were not in agreement with IAH. Hence the model is experimentally supported only in pathosystems in which the host is highly susceptible. We provide, to our knowledge, the first qualitative experimental evidence that, in such pathosystems, the action of a single virion is sufficient to cause disease.

Host plant species can influence the fitness of herbivore pathogens: the winter moth and its nucleopolyhedrovirus.

Plants can have a significant impact on the fitness and efficacy of natural enemies. These interactions are widespread and suggest that the influences on the population dynamics of insect herbivores cannot be simply divided into bottom up and top down. Several questions remain little studied in this field. Firstly, to what extent can plants affect the interactions between insects and their pathogens? Secondly, what are the effects of variation within natural enemy species on host/enemy/plant interactions? Finally, if plant/pathogen interactions can occur, do pathogens have increased fitness on the locally abundant food plant of their host? This study explored the influence of three host plant species of the polyphagous winter moth, Operophtera brumata, on infections caused by two geographic isolates of the winter moth nucleopolyhedrovirus (NPV) collected from distinct winter moth habitats. Insects were infected on excised leaf tissue of common oak, Quercus robur, Sitka spruce, Picea sitchenis, and heather, Calluna vulgaris. Parameters fundamental to the basic reproductive rate of the pathogen were estimated: these being infectivity, speed of kill and the yield of virus per insect. Leaf nitrogen and phenolic content were measured as indicators of host plant quality for the three plant species: oak had the highest levels of nitrogen and also the highest levels of phenolic compounds. Heather had higher levels of phenolic compounds than Sitka spruce. Host plant did not affect the infectivity of either isolate but insects that ingested virus on oak foliage died sooner and yielded more virus than insects that ingested virus on Sitka spruce or heather. The effect of host plant species on pathogen yield varied between the two isolates of the NPV but not as predicted by our adaptive hypothesis. The interactions between virus and food plant are discussed in relation to host and pathogen population dynamics.

TRANSMISSION PATTERNS OF NATURAL AND RECOMBINANT BACULOVIRUSES

The advent of genetically modified organisms such as pathogens has raised ecological questions that need to be addressed in order to assess any risks involved in their use. The baculovirus Autographa californica nucleopolyhedrovirus (AcNPV), which infects a number of lepidopteran species, has been modified to express an insect-selective toxin. This genetic modification increases the speed with which it kills its host. However, in addition to this intended feature of the modified virus, there may be other consequences for the host-pathogen interaction. We report a field experiment in which transmission patterns of the wild-type and the genetically modified baculovirus are measured within and between a model target (susceptible) and nontarget (less susceptible) lepidopteran species. Two foliar feeders were chosen: Trichoplusia ni, the cabbage looper, is highly susceptible to this pathogen, while Mamestra brassicae, the cabbage moth, is semipermissive. These two species are used as both the source and the recipients of infection for both virus types. A series of models are fitted to determine the probabilities of infection (given survival from other sources of mortality) over a 7-d period within contained field cages. Fitting these models to data illustrates that a substantial fraction of the population escapes infection, and it is the size of the pathogen-free refuge that varies between treatments. When infected individuals from the less susceptible species die, the yield of virus is greater than from susceptible hosts, yet this does not significantly alter the risk of transmission to other hosts. In contrast, the genetically modified baculovirus always results in a lower risk of infection in the field compared to the wild type. This is because the recombinant virus causes paralysis, and as a result, the cadaver may fall from the plant before death and virus release. Hence the number of cadavers remaining on the foliage has a greater influence on transmission than the yield of virus from those cadavers.

Hierarchical spatial structure of genetically variable nucleopolyhedroviruses infecting cyclic populations of western tent caterpillars.

The cyclic population dynamics of western tent caterpillars, Malacosoma californicum pluviale, are associated with epizootics of a nucleopolyhedrovirus, McplNPV. Given the dynamic fluctuations in host abundance and levels of viral infection, host resistance and virus virulence might be expected to change during different phases of the cycle. As a first step in determining if McplNPV virulence and population structure change with host density, we used restriction fragment length polymorphism (RFLP) analysis to examine the genetic diversity of McplNPV infecting western tent caterpillar populations at different spatial scales. Thirteen dominant genetic variants were identified in 39 virus isolates (individual larvae) collected from field populations during one year of low host density, and another distinct variant was discovered among nine additional isolates in two subsequent years of declining host density. The distribution of these genetic variants was not random and indicated that the McplNPV population was structured at several spatial levels. A high proportion of the variation could be explained by family grouping, which suggested that isolates collected within a family were more likely to be the same than isolates compared among populations. Additionally, virus variants from within populations (sites) were more likely to be the same than isolates collected from tent caterpillar populations on different islands. This may indicate that there is limited mixing of virus among tent caterpillar families and populations when host population density is low. Thus there is potential for the virus to become locally adapted to western tent caterpillar populations in different sites. However, no dominant genotype was observed at any site. Whether and how selection acts on the genetically diverse nucleopolyhedrovirus populations as host density changes will be investigated over the next cycle of tent caterpillar populations.

Behavior of a recombinant baculovirus in lepidopteran hosts with different susceptibilities.

ABSTRACT Insect pathogens, such as baculoviruses, that are used as microbial insecticides have been genetically modified to increase their speed of action. Nontarget species will often be exposed to these pathogens, and it is important to know the consequences of infection in hosts across the whole spectrum of susceptibility. Two key parameters, speed of kill and pathogen yield, are compared here for two baculoviruses, a wild-type Autographa californica nucleopolyhedrovirus (AcNPV), AcNPV clone C6, and a genetically modified AcNPV which expresses an insect-selective toxin, AcNPV-ST3, for two lepidopteran hosts which differ in susceptibility. The pathogenicity of the two viruses was equal in the less-susceptible host, Mamestra brassicae, but the recombinant was more pathogenic than the wild-type virus in the susceptible species, Trichoplusia ni. Both viruses took longer to kill the larvae of M. brassicae than to kill those of T. ni. However, whereas the larvae of T. ni were killed more quickly by the recombinant virus, the reverse was found to be true for the larvae ofM. brassicae. Both viruses produced a greater yield inM. brassicae, and the yield of the recombinant was significantly lower than that of the wild type in both species. The virus yield increased linearly with the time taken for the insects to die. However, despite the more rapid speed of kill of the wild-type AcNPV in M. brassicae, the yield was significantly lower for the recombinant virus at any given time to death. A lower yield for the recombinant virus could be the result of a reduction in replication rate. This was investigated by comparing determinations of the virus yield per unit of weight of insect cadaver. The response of the two species (to both viruses) was very different: the yield per unit of weight decreased over time for M. brassicae but increased for T. ni. The implications of these data for risk assessment of wild-type and genetically modified baculoviruses are discussed.

Evolution and the microbial control of insects

Insect pathogens can be utilized in a variety of pest management approaches, from inundative release to augmentation and classical biological control, and microevolution and the consideration of evolutionary principles can potentially influence the success of all these strategies. Considerable diversity exists in natural entomopathogen populations and this diversity can be either beneficial or detrimental for pest suppression, depending on the pathogen and its mode of competition, and this should be considered in the selection of isolates for biological control. Target hosts can exhibit considerable variation in their susceptibility to entomopathogens, and cases of field‐evolved resistance have been documented for Bacillus thuringiensis and baculoviruses. Strong selection, limited pathogen diversity, reduced gene flow, and host plant chemistry are linked to cases of resistance and should be considered when developing resistance management strategies. Pre‐ and post‐release monitoring of microbial control programs have received little attention; however, to date there have been no reports of host‐range evolution or long‐term negative effects on nontarget hosts. Comparative analyses of pathogen population structure, virulence, and host resistance over time are required to elucidate the evolutionary dynamics of microbial control systems.