Nutrition as a neglected factor in insect herbivore susceptibility to Bt toxins.

The widespread global adoption of Bt crops elevates concerns about the evolution of Bt resistance in insect pest species. Current insecticide resistance management (IRM) strategies focus solely on genetic variation as a causal factor in the evolution of resistance, but ignore the role that environmental factors, such as nutrition, may play. In this opinion paper, we discuss the benefits that insect herbivores gain from consuming foods with protein-carbohydrate content that matches their self-selected protein-carbohydrate intake, and show that even within monocultures there is amply opportunity for insect herbivores to regulate their macronutrient intake. Next we review new data that show that dietary protein and carbohydrates can: firstly, have predictably strong effects on the survival and performance of caterpillars challenged with Bt toxins, and secondly, mediate plasticity in susceptibility to Cry1Ac, which can account for large differences in LC50 values. Nutrition-Bt interactions such as these have important implications for IRM, particularly given that diet-incorporated Bt bioassays commonly use artificial diets that vary substantially from their self-selected optimal diets, which likely results in underestimates of resistance in the field. Failing to bioassay larvae on ecologically-relevant diets can seriously confound the results of Bt resistance monitoring bioassays and undermine our ability to detect resistance in the field.

Alternative migratory locust phenotypes are associated with differences in the expression of genes encoding the methylation machinery.

Despite the importance of locust density-dependent polyphenism as a model system for understanding phenotypic plasticity, there is still much to be learnt about its underlying molecular control. Here we describe the first investigation into the expression of genes encoding the DNA methylation machinery in the migratory locust (Locusta migratoria). We show that the alternative solitarious and gregarious phenotypic states induced by different locust rearing densities are associated with significant differences in the expression of the target genes DNA methyltransferase 1, DNA methyltransferase 2 and methyl-CpG-binding domain protein 2/3. This variation was most pronounced in the embryos of solitarious vs. gregarious mothers. We mapped the embryonic methylation profiles of several intragenic regions and a Long Interspersed Nuclear Element (LINE), each of which is known to be differentially expressed between alternative locust phenotypes or has been directly implicated in phase change. LmI and three genes, adenyl cyclase-associated binding protein 2, choline kinase alpha-like and henna, were methylated. Our results set the stage for future studies investigating the specific role of DNA methylation in the maternal transfer of migratory locust phase polyphenism.

Long microsatellites and unusually high levels of genetic diversity in the Orthoptera.

Much remains to be learned about the mutational processes governing the evolution of microsatellite repeat regions and the associated levels of genetic diversity observed at microsatellite markers across populations or species. An extensive survey of microsatellite variation in 210 insect species from six major orders revealed that within Orthopterans, which are characterized by giant genomes, levels of genetic diversity were ~20% higher and microsatellite repeat arrays were longer than in any other group. Because of the mutation dependence on repeat length, this result suggests a higher microsatellite loci mutation rate in the Orthoptera. We deem it plausible that differences among insect orders, either in mismatch repair systems or in abundance of transposable element-derived microsatellites, can shape the size distribution of both genomes and microsatellite repeat regions. Our findings emphasise that observed levels of genetic diversity can greatly vary across species (orders at least) because of molecular differences in the mechanisms that determine microsatellite size, and are therefore critical to conservation and population genetics studies, where microsatellite repeat variability is primarily interpreted in terms of population demography and history.

Radiotelemetric analysis of the effects of prevailing wind direction on Mormon cricket migratory band movement.

During outbreaks, flightless Mormon crickets [Anabrus simplex Haldeman (Orthoptera: Tettigoniidae)] form large mobile groups known as migratory bands. These bands can contain millions of individuals that march en masse across the landscape. The role of environmental cues in influencing the movement direction of migratory bands is poorly understood and has been the subject of little empirical study. We examined the effect of wind direction on Mormon cricket migratory band movement direction by monitoring the local weather conditions and daily movement patterns of individual insects traveling in bands over the same time course at three close, but spatially distinct sites. Although weather conditions were relatively homogeneous across sites, wind directions tended to be more variable across sites during the morning hours, the period during which directional movement begins. Migratory bands at different sites traveled in distinctly different directions. However, we failed to find any evidence to suggest that the observed variation in migratory band movement direction was correlated with local wind direction at any time during the day. These results support the notion that the cues mediating migratory band directionality are likely to be group specific and that a role for landscape-scale environmental cues such as wind direction is unlikely.

Do outbreaks affect genetic population structure? A worldwide survey in Locusta migratoria, a pest plagued by microsatellite null alleles.

An understanding of the role of factors intrinsic to a species' life history in structuring contemporary genetic variation is a fundamental, but understudied, aspect of evolutionary biology. Here, we assessed the influence of the propensity to outbreak in shaping worldwide genetic variation in Locusta migratoria, a cosmopolitan pest well known for its expression of density-dependent phase polyphenism. We scored 14 microsatellites in nine subspecies from 25 populations distributed over most of the species' range in regions that vary in the historical frequency and extent of their outbreaks. We rejected the hypothesis that L. migratoria consists of two genetically distinct clusters adapted to habitats either rarely (nonoutbreaking) or cyclically (outbreaking) favourable to increases in population density. We also invalidated the current subspecific taxonomic classification based on morphometrics. Bayesian inferences indicated evidence of a homogenizing effect of outbreaks on L. migratoria population structure. Geographical and ecological barriers to gene flow in conjunction with historical events can also explain the observed patterns. By systematically assessing the effects of null alleles using computer simulations, we also provide a template for the analysis of microsatellite data sets characterized by a high prevalence of null alleles.

Ancient trans-Atlantic flight explains locust biogeography: molecular phylogenetics of Schistocerca.

The desert locust (Schistocerca gregaria) has been an important agricultural pest at least since biblical times. Although the ecology, physiology and behaviour of this insect species have been well characterized, its biogeographical origins and evolutionary history are more obscure. Schistocerca gregaria occurs throughout Africa, the Middle East and Western Asia, but all other species in the genus Schistocerca are found in the New World. Because S. gregaria has the capacity for extreme long-distance movement associated with swarming behaviour, dispersal may have played an important role in determining current distribution patterns. Some authors have argued that S. gregaria is the product of an eastward trans-Atlantic dispersal from North America to Africa; others consider it more likely that the New World taxa are the product of westward dispersal from Africa. Here, we present a mitochondrial DNA phylogeny of Schistocerca species that supports the monophyly of New World species (including the Galapagos endemic Halmenus) relative to S. gregaria. In concert with observed patterns of molecular divergence, and in contrast to previous morphological studies, our analysis indicates a single trans-Atlantic flight from Africa to South America, followed by extensive speciation and ecological divergence in the New World.

Host plant-associated genetic differentiation in the snakeweed grasshopper, Hesperotettix viridis (Orthoptera: Acrididae).

Studies of herbivorous insects have played a major role in understanding how ecological divergence can facilitate genetic differentiation. In contrast to the majority of herbivorous insects, grasshoppers as a group are largely polyphagous. Due to this relative lack of intimate grasshopper-plant associations, grasshopper-plant systems have not played a large part in the study of host-associated genetic differentiation. The oligophagous grasshopper, Hesperotettix viridis (Thomas), is endemic to North America and feeds on composites (Asteraceae) within the tribe Astereae. Previous work has shown both preference and performance differences between H. viridis individuals feeding on either Solidago mollis or Gutierrezia sarothrae. Using 222 AFLP markers, we examined the genetic relationships among 38 H. viridis individuals feeding on these plants both in sympatry and allopatry. Neighbour-joining analysis resulted in two distinct host-associated clades with 71% bootstrap support for host-associated monophyly. Analyses of molecular variation (amova) revealed significant genetic structuring with host plant accounting for 20% of the total genetic variance while locality accounted for 0%. Significant genetic differentiation was detected between S. mollis-feeders and G. sarothrae-feeders even when the two were present at the same locality. These results are consistent with observed differences in preference and performance between H. viridis grasshoppers feeding on either G. sarothrae or S. mollis and indicate that H. viridis is comprised of at least two genetically distinct host plant-associated lineages.

Density-dependent aposematism in the desert locust.

The ecological processes underlying locust swarm formation are poorly understood. Locust species exhibit phenotypic plasticity in numerous morphological, physiological and behavioural traits as their population density increases. These density-dependent changes are commonly assumed to be adaptations for migration under heterogeneous environmental conditions. Here we demonstrate that density-dependent nymphal colour change in the desert locust Schistocerca gregaria (Orthoptera: Acrididae) results in warning coloration (aposematism) when the population density increases and locusts consume native, toxic host plants. Fringe-toed lizards (Acanthodactylus dumerili (Lacertidae)) developed aversions to high-density-reared (gregarious-phase) locusts fed Hyoscyamus muticus (Solanaceae). Lizards associated both olfactory and visual cues with locust unpalatability, but only gregarious-phase coloration was an effective visual warning signal. The lizards did not associate low rearing density coloration (solitarious phase) with locust toxicity. Predator learning of density-dependent warning coloration results in a marked decrease in predation on locusts and may directly contribute to outbreaks of this notorious pest.