The relationship between the degree of specialization of parasitoids and their responses to host-related volatiles is an important and current evolutionary question. Specialist parasitoids which have evolved to attack fewer host species are predicted to be more responsive to host-related volatiles than generalists. We tested the above prediction by comparing behavioural responses of both sexes of two parasitoids (Hymenoptera: Braconidae) with different degrees of host specificity, Microplitis croceipes (Cresson) (specialist) and Cotesia marginiventris (generalist), to different suites of synthetic host-related volatile compounds. The compounds tested at two doses (1 and 100μg) include two green leaf volatiles (GLVs: hexanal and (Z)-3-hexen-1-ol) and four herbivore-induced plant volatiles (HIPVs: (Z)-3-hexenyl acetate, linalool, (Z)-3-hexenyl butyrate and (E,E)-α-farnesene). Two hypotheses were tested: (i) M. croceipes (specialist) would show relatively greater behavioural responses to the HIPVs, whereas C. marginiventris (generalist) would show greater behavioural responses to the GLVs, and (ii) females of both species would show greater responses than conspecific males to the host-related volatiles. At the low dose (1μg), females of the specialist showed significantly greater responses than females of the generalist to three of the tested HIPVs, (Z)-3-hexenyl acetate, linalool and (Z)-3-hexenyl butyrate. In contrast, females of the generalist showed relatively greater responses to the GLVs. The same trends were recorded at the high dose but fewer significant differences were detected. In general, similar results were recorded for males, with the exception of linalool (an HIPV) which elicited significantly greater response in the generalist than the specialist. Comparing the sexes, females of both species showed greater responses than conspecific males to most of the tested volatiles. The ecological significance of these findings is discussed.

The red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae), is an economically important pest of palms worldwide. To better understand the phenology of R. ferrugineus populations in the Mediterranean basin, the objective of the present work has been to estimate the effect of temperatures within the range 10–25°C on the reproductive parameters of R. ferrugineus and to determine its lower temperature thresholds for oviposition and egg hatching. Our results confirm that oviposition in R. ferrugineus is strongly affected by temperature. Lower thresholds for oviposition and egg hatching (15.45° and 13.95°C, respectively) are below the mean monthly temperatures registered in winter in most of the northern shore of the Mediterranean basin. Under these circumstances, new palm infestations would be difficult during most of the winter. These results should be taken into account when planning some palm management practices, such as pruning or pesticide treatments, in areas under a Mediterranean climate.

Thrips are small insects (0.5–3.0 mm) with distinct habits and life histories characterized by haplodiploid sex determination. In general, low levels of genetic diversity have been reported in haplodiploid insects, although most reports focus on the order Hymenoptera. Therefore, we used RAPD markers to evaluate the structure and both inter- and intra-population genetic variability of Gynaikothrips uzeli (Thysanoptera: Phlaeothripidae). Six populations, three from Paraná state, southern Brazil, and three from Bahia, northeastern Brazil, were studied. Similarly to other haplodiploid insects, the genetic diversity of G. uzeli was reduced. This result is putatively related to the haplodiploid sex determination system, which yields little genetic variation, and to ecological traits of the studied species, such as the low dispersal abilities and life mode in leaf galls. All individuals were homogeneously clustered in their respective collection sites, forming two main groups in which populations from similar environments were more closely related. The analyzed populations were highly structured, and the genetic variation was higher among than within populations.

Cotton growing landscapes in Australia have been dominated by dual-toxin transgenic Bt varieties since 2004. The cotton crop has thus effectively become a sink for the main target pest, Helicoverpa armigera. Theory predicts that there should be strong selection on female moths to avoid laying on such plants. We assessed oviposition, collected from two cotton-growing regions, by female moths when given a choice of tobacco, cotton and cabbage. Earlier work in the 1980s and 1990s on populations from the same geographic locations indicated these hosts were on average ranked as high, mid and low preference plants, respectively, and that host rankings had a heritable component. In the present study, we found no change in the relative ranking of hosts by females, with most eggs being laid on tobacco, then cotton and least on cabbage. As in earlier work, some females laid most eggs on cotton and aspects of oviposition behaviour had a heritable component. Certainly, cotton is not avoided as a host, and the implications of these finding for managing resistance to Bt cotton are discussed.

Pieris brassicae (Linn.) is a destructive cosmopolitan pest of cruciferous crops. It is present wherever its host plants occur, and it is considered to be one of the most widely distributed of all the Lepidoptera. We investigated the affect of various host plants on the food consumption and utilization by P. brassicae. We quantified consumption of food, larval duration, pupal duration and weight on cabbage (Brassica oleracea var. capitata), cauliflower (Brassica oleracea var. botrytis), radish (Raphanus sativus), broccoli (Brassica oleracea var. italica) and mustard (Brassica campestris) under laboratory conditions. Insect-host relationships can be better understood by knowing the rate of food consumption, its digestibility and conversion of food eaten to body tissue. The consumption of food generally increased with the advancement of larval age. In our study we found that consumption of food was highest on radish and lowest on broccoli. The highest consumption of a particular host does not always indicate greater suitability of that host, until and unless other factors like consumption index (CI), relative growth rate (RGR), efficiency of conversion of ingested food (ECI), approximate digestibility (AD) and efficiency of conversion of digested food (ECD) are also considered. In the current investigation, factors like CI, RGR, ECI and ECD were highest on cabbage. Low body weight of pupa is associated with rapid development. On cabbage, the weight of pupa of both sexes was found lowest. Thus, from the present study, it can be concluded that cabbage is a more suitable host for P. brassicae than other host plants evaluated. Hence, on cabbage, the values of Waldbauer indices were highest and P. brassicae developed with a faster rate.

Understanding how the population dynamics of insect pests are affected by environmental factors and agricultural practices is important for pest management. To investigate how the abundance of the banana weevil, Cosmopolites sordidus (Coleoptera: Curculionidae), is related to environmental factors and the length of the fallow period in Martinique, we developed an extensive data set (18,130 observations of weevil abundance obtained with pheromone traps plus associated environmental data) and analysed it with generalized mixed-effects models.

At the island scale, C. sordidus abundance was positively related to mean temperature and negatively related to mean rainfall but was not related to soil type. The number of insects trapped was highest during the driest months of the year. Abundance of C. sordidus decreased as the duration of the preceding fallow period increased.

The latter finding is inconsistent with the view that fallow-generated decomposing banana tissue is an important resource for larvae that leads to an increase in the pest population. The results are consistent with the view that fallows, in association with pheromone traps, are effective for the control of the banana weevil.

The legume pod borer, Maruca vitrata, is an endemic insect pest that causes significant yield loss to the cowpea crop in West Africa. The application of population genetic tools is important in the management of insect pests but such data on M. vitrata is lacking. We applied a set of six microsatellite markers to assess the population structure of M. vitrata collected at five sites from Burkina Faso, Niger and Nigeria. Observed polymorphisms ranged from one (marker 3393) to eight (marker 32008) alleles per locus. Observed and expected heterozygosities ranged from 0.0 to 0.8 and 0.0 to 0.6, respectively. Three of the loci in samples from Nigeria and Burkina Faso deviated significantly from Hardy-Weinberg Equilibrium (HWE), whereas no loci deviated significantly in samples from Niger. Analysis of molecular variance (AMOVA) indicated that 67.3% level of the genetic variation was within individuals compared to 17.3% among populations. A global estimate of FST=0.1 (ENA corrected FST=0.1) was significant (P⩽0.05) and corroborated by pairwise FST values that were significant among all possible comparisons. A significant correlation was predicted between genetic divergence and geographic distance between subpopulations (R2=0.6, P=0.04), and cluster analysis by the program STRUCTURE predicted that co-ancestry of genotypes were indicative of three distinct populations. The spatial genetic variance among M. vitrata in West Africa may be due to limited gene flow, south-north seasonal movement pattern or other reproductive barriers. This information is important for the cultural, chemical and biological control strategies for managing M. vitrata.

We investigated the molecular diversity of the major agricultural pest Bemisia tabaci and of its associated secondary endosymbionts in Greece. Analyzing mitochondrial DNA, we found that the Q1 (=Q west) is predominant. We used eight microsatellite polymorphic markers to study the genetic structure of 37 populations from mainland and insular Greece, collected on different host species from outdoor and protected crops as well as from non-cultivated plants. In some cases, gene flow was found to be low even between populations separated by just a few kilometres. Bayesian analysis identified two main genetic groups, the first encompassing populations from south Crete and the second composed of populations from north Crete, two other Aegean islands and mainland Greece. Genetic differentiation was not correlated with different host plant species or habitat, or greenhouse versus open environment populations. Gene flow significantly decreased with geographic distance, but no isolation by distance existed when only the samples from mainland Greece or only the samples from Crete were considered. The secondary symbionts Wolbachia and Hamiltonella were present at high frequencies while Arsenophonus, Cardinium and Rickettsia were absent from Greek populations. Multilocus sequence typing of Wolbachia identified two Wolbachia strains. These two strains were found together in most of the populations studied but never in the same host individual. Their role on the observed population structure is discussed.

It is important to understand how components of the agroecosystem interfere with the attack of a pest species and their seasonality in order to use these components in IPM programs. This study focused on the evaluation of the seasonality of natural control factors associated with the guava psyllid Triozoida limbata (Enderlein) in Brazil. Life-table data were collected from an experimental guava orchard during four periods that roughly represented four seasons. Natural mortality was monitored daily through the immature stages, and the relative importance of each natural mortality factor and its seasonality was determined. Significant statistical differences were observed in the mortality during the four periods (P < 0.05). Several factors contributed to the mortality of T. limbata, including rainfall, physiological disturbance, the parasitoid Psyllaephagus sp. (Hymenoptera: Encyrtidae) and specific predators: syrphids, predatory wasps and other generalist predators. Depending on the location of nymphs (exposed or inside galls), the relative importance of the different natural mortality factors changed. The principal component analysis (PCA) showed some trends in the relationship of natural control agents and weather conditions. For example, the occurrence of predatory wasps was positively correlated with temperature and occurrence of winds; the occurrence of syrphids and Psyllaephagus sp. were negatively correlated with temperature and winds; and the occurrence of other generalist predators were negatively correlated with the occurrence of rainfall and photoperiod. The results showed the importance of natural mortality factors for the management of T. limbata and their changes through the different seasons which should be considered when implementing IPM programs in guava orchards.

Four morphologically cryptic species of the Bactrocera dorsalis fruit fly complex (B. dorsalis s.s., B. papayae, B. carambolae and B. philippinensis) are serious agricultural pests. As they are difficult to diagnose using traditional taxonomic techniques, we examined the potential for geometric morphometric analysis of wing size and shape to discriminate between them. Fifteen wing landmarks generated size and shape data for 245 specimens for subsequent comparisons among three geographically distinct samples of each species. Intraspecific wing size was significantly different within samples of B. carambolae and B. dorsalis s.s. but not within samples of B. papayae or B. philippinensis. Although B. papayae had the smallest wings (average centroid size=6.002 mm±0.061 SE) and B. dorsalis s.s. the largest (6.349 mm±0.066 SE), interspecific wing size comparisons were generally non-informative and incapable of discriminating species. Contrary to the wing size data, canonical variate analysis based on wing shape data discriminated all species with a relatively high degree of accuracy; individuals were correctly reassigned to their respective species on average 93.27% of the time. A single sample group of B. carambolae from locality ‘TN Malaysia’ was the only sample to be considerably different from its conspecific groups with regards to both wing size and wing shape. This sample was subsequently deemed to have been originally misidentified and likely represents an undescribed species. We demonstrate that geometric morphometric techniques analysing wing shape represent a promising approach for discriminating between morphologically cryptic taxa of the B. dorsalis species complex.

Generalist predators and parasitoids are considered to be important regulators of aphids. The former not only feed on these pests, but might also consume parasitoids at all stages of development. This direct or coincidental interference affects the natural control of aphids, the scale of which is largely unknown, and it has rarely been examined under natural conditions. Here, molecular diagnostics were used to track trophic interactions in an aphid-parasitoid-generalist predator community during the build-up of a cereal aphid population. We found that generalist predators, principally carabid and staphylinid beetles as well as linyphiid spiders, had strong trophic links to both parasitoids and aphids. Remarkably, more than 50% of the parasitoid DNA detected in predators stems from direct predation on adult parasitoids. The data also suggest that coincidental intraguild predation is common too. Generalist predators, hence, disrupt parasitoid aphid control, although the levels at which the predators feed on pests and parasitoids seem to vary significantly between predator taxa. Our results suggest that taxon-specific trophic interactions between natural enemies need to be considered to obtain a more complete understanding of the route to effective conservation biological control.

Phosphoglucose isomerase (PGI) plays an important role in energy metabolism, and it is documented that PGI exhibits an extensive polymorphism which can affect insects' fitness and adaptation. In this paper, we studied the structural characteristics and polymorphism of pgi gene in the fall webworm (Hyphantria cunea), an important invasive pest in some European and Asian countries. A 2110-bp pgi full-length cDNA encoding a polypeptide of 556 amino acids was obtained from H. cunea. The pgi full-length in the H. cunea genomic DNA was 14,332 bp with 12 exons and 11 introns, similar to the structures of pgi in other Lepidoptera species. We compared the structures of pgi in different insect species. Moreover, thirteen pgi genotypes comprised of five alleles were identified in the Chinese population. Genotypes pgi-cd, pgi-cc and pgi-ce were the most prevalent with over 70% of individuals allocated to them. Four out of five alleles were sequenced the cDNA full-length. Thirty stably variable sites were found among them with five non-synonymous mutation sites. The frequencies of alleles and genotypes were variable in different Chinese geographic subpopulations. Moreover, comparison of pgi mRNA expression levels in each stage of the moth's lifecycle showed that a high expression level was in the 6th instar larval stage, followed by that in the egg and adult stages. The results will provide a basis for further study of the role of different alleles and genotypes of PGI on fitness and adaptation of the moth H. cunea.

Temperature is considered to be an important abiotic factor influencing insect reproduction. Despite the importance of Plutella xylostella L. (Lepidoptera: Plutellidae) as a pest of brassicaceous crops worldwide, the effects of temperature on its reproduction are not well understood. We evaluated the effect of constant temperatures ranging from 10 to 32.5°C on the reproduction of P. xylostella and developed an oviposition model for the species. The model combined temperature-dependent parameters of total fecundity, age-specific oviposition rate and age-specific survival. Additionally, we modelled population growth as a function of temperature. The estimated parameters allowed us to discuss the possible consequences of global warming on P. xylostella distribution. Temperature affected the length of pre-oviposition after adult emergence, oviposition period, longevity, total fecundity and egg viability. The model predicted that both daily egg production and length of oviposition period decreased at temperatures below 15°C and above 25°C. Population growth increased linearly with temperature in a range from 10°C to 25°C; however, the model predicted a reduction in population growth at temperatures above 28.6°C. Data suggested that temperature plays a critical role in P. xylostella reproduction, and subtle differences in average temperature could have an impact on its population growth. This is especially important in the context of global climate change, which in turn could alter the distribution and abundance of the pest in some regions of the world.

The females of the white grub beetle, Dasylepida ishigakiensis, release two enantiomers of 2-butanol, (R)-2-butanol and (S)-2-butanol. The ratio describing the relative proportions of these two enantiomers (R/S ratio) has not yet been investigated. (R)-2-Butanol has been shown to attract males in laboratory and field experiments, whereas (S)-2-butanol tends to inhibit them. To determine the R/S ratio of the 2-butanol emitted by virgin females, we collected 2-butanol from young (53 days old), mature (63 days old) and old females (73 days old) using water, extracted with an SPME fibre and subsequently injected into GC-MS. The major component of the 2-butanol emitted by the young females was (R)-2-butanol, but as the females aged, the component ratio favoured (S)-2-butanol. Young females released an 80:20 mixture of (R)- and (S)-2-butanol, whereas old females released a 45:55 mixture. The EAG response of male antennae to a 50:50 ratio (racemic mixture) showed a similar dose-response curve to that of (R)-2-butanol. The male orientation responses to (R)-2-butanol decreased when the relative proportion of (S)-2-butanol increased. An inhibitory and/or masking effect of (S)-2-butanol on male orientation behaviour was also observed in the flight tunnel assay. These results suggest that males are more strongly attracted to young females than to old females. We also discuss the possibility of using 2-butanol isomers as a control or monitoring agent for this insect.

Insect chemosensory proteins (CSPs) are supposed to transport hydrophobic chemicals to receptors on sensory neurons. However, CSPs are broadly expressed in various insect tissues, suggesting their involvement in the physiological processes beyond chemoreception. So, the exact physiological roles of CSPs in insects still need to be unraveled. In this study, three full-length of CSP genes from Spodoptera exigua have been cloned and characterized. The deduced amino acid sequences of SexiCSP1, SexiCSP2 and SexiCSP3 revealed open reading frames of 128, 128 and 126 amino acids, respectively, with four conserved cysteine residues. The expression patterns of the three SexiCSPs were further investigated by real-time PCR. Three SexiCSPs were expressed in antennae, heads, legs, wings, thoraxes, abdomens, testes and ovaries, with the highest expression level in female and male antennae. Furthermore, all three SexiCSPs mRNA were distributed extensively in the tested development stages with the highest expression level in pupae. RNAi-based gene silencing study resulted in a dramatic reduction of corresponding mRNA in female S. exigua after injection with dsRNA of all three SexiCSPs. Consequentially, 42.5% of mortalities, 68.3% (compare to DEPC water injected control) and 71.4% (compare to uninjected control) oviposition inhibition, and significantly effected egg hatching were observed in the female S. exigua injected with dsSexiCSP3 as compared to control treatments. On the other hand, dsSexiCSP1 and dsSexiCSP2 injected female adults did not show effects on survival and reproduction. Our study confirms the utility of RNAi approach to functional characterization of CSP genes in S. exigua and provides a starting point for further studies on female survival and reproduction in this insect. It also reveals the potential pest controlling method, as insect behavior regulation agent that disrupts the expression of chemosensory proteins.

To assess genetic diversity in populations of the brown planthopper (Nilaparvata lugens Stål) (Homoptera: Delphacidae), we have developed and applied microsatellite, or simple sequence repeat (SSR), markers from expressed sequence tags (ESTs). We found that the brown planthopper clusters of ESTs were rich in SSRs with unique frequencies and distributions of SSR motifs. Three hundred and fifty-one EST-SSR markers were developed and yielded clear bands from samples of four brown planthopper populations. High cross-species transferability of these markers was detected in the closely related planthopper N. muiri. The newly developed EST-SSR markers provided sufficient resolution to distinguish within and among biotypes. Analyses based on SSR data revealed host resistance-based genetic differentiation among different brown planthopper populations; the genetic diversity of populations feeding on susceptible rice varieties was lower than that of populations feeding on resistant rice varieties. This is the first large-scale development of brown planthopper SSR markers, which will be useful for future molecular genetics and genomics studies of this serious agricultural pest.

Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) develops in chestnut buds that remain asymptomatic from oviposition (June–July) until budburst; it is, thus, easily spread by plant material used in propagation. Therefore, it is particularly interesting to identify infested plant batches before their movement. Unfortunately, a non-destructive method for checking buds has not yet been developed, and the only technique available is the screening of a bud sample. The visual investigation is long and requires highly skilled and trained staff. The purpose of this work was to set up an effective and fast method able to identify the presence of first instar larvae of D. kuriphilus in a large number of chestnut buds by PCR. Four primer pairs were designed on nuclear and mitochondrial sequences of a set of seven gall wasp taxa and tested on five different cynipid's DNA. Nested diagnostic PCR was carried out on DNA extracted from samples of 2 g buds simulating four levels of infestation (larvae were added to uninfested buds); 320 bp amplicon of 28S sequence was chosen as a marker to detect one larva out of 2 g buds. The method showed a potential efficiency of 5000 to 15,000 buds per week, depending on bud size.

The insect cuticle is covered by a thin layer of hydrocarbons not only preventing desiccation but also playing an important role in the sexual communication of several species. In the pteromalid wasp Lariophagus distinguendus, a parasitoid of grain infesting beetles, female cuticular hydrocarbons (CHCs) elicit male courtship behaviour. We analyzed the CHC profiles of male and female L. distinguendus wasps reared on different beetle hosts by coupled gas chromatography- mass spectrometry (GC-MS). Statistical analysis of the data revealed significant differences between strains reared on different hosts, while spatially isolated strains reared on the same host produced similar profiles. CHC profiles of parasitoids reared on Stegobium paniceum were statistically distinguishable from those of wasps reared on all other hosts. A host shift from Sitophilus granarius to S. paniceum resulted in distinguishable CHC profiles of L. distinguendus females after only one generation. Considering the role of CHCs as contact sex pheromones, our data suggest that host shifts in parasitic wasps might lead to reproductive isolation of host races due to the modification of the cuticular semiochemistry.