Some bark beetle species (Coleoptera: Scolytinae) produce aggregation pheromones that allow coordinated attack on their conifer hosts. As a new saproxylic habitat is founded, an assemblage of associated beetles kairomonally respond to bark beetle infochemicals. Ips sexdentatus is one of the major damaging insects of Pinus spp. in Southern Europe. Its response to varying ipsenol (Ie) percentages in relation to ipsdienol (Id) was studied in northwestern Spain, along with the entire saproxylic beetle assemblage captured at multiple-funnel traps. Response profile modeling was undertaken for I.sexdentatus sexes and sex-ratios, associated species and for selected trophic groups using a reference Gaussian model. In addition, the effects on the saproxylic assemblages were analyzed. I. sexdentatus response curve peaked at 22.7% Ie content, while remaining taxa that could be modeled, peaked above ca. 40% Ie. Predator guilds showed a linear relationship with Ie proportion, while competitors showed a delayed response peak. Consequently, species assemblages differed markedly between varying pheromone component mixtures. Given that the evaluated pheromonal proportions mimicked that of logs being colonized by I. sexdentatus, results suggested that the registered differential responses at different levels might provide I.sexdentatus with a temporal window that maximizes conspecific attraction while reducing interference with competitor and predatory guilds. Described responses might help improve the monitoring of the population status of target bark beetles and their associates, but also point toward the by-catch of many natural enemies, as well as rare saproxylic beetle species, interfering with the aims of sustainable forest management.

The black currant-lettuce aphid, Nasonovia ribisnigri, is an important pest of cultivated lettuce, Lactuca sativa. Since 1982, the control of this aphid on lettuce is largely based on host plant resistance, conferred by the Nr gene, introgressed from Lactuca virosa. The resistance mechanism remains to be identified. N. ribisnigri populations virulent on the Nr-based resistance in lettuce have emerged in several locations in Europe since 2007. The objective of this study was to investigate the resistance mechanism mediated by the Nr gene in lettuce by detailed studies of aphid feeding behaviour and performance. Both avirulent (Nr:0) and virulent (Nr:1) biotypes of N. ribisnigri were studied on five resistant and two susceptible near isogenic lines (NILs). In addition, survival and colony development were quantified. Nr:0 aphids showed a strong decrease in sieve element ingestion and took longer to accept a sieve element on resistant NILs compared with susceptible NILs, and no aphids survived on the resistant NIL. Nr:1 aphids fed and performed equally well on the resistant and susceptible NILs. The resistance mechanism against Nr:0 aphids encoded by the Nr gene seems to be located in the phloem, although we also observed differences in feeding behaviour during the pathway phase to the phloem. Nr:1 aphids were highly virulent to the resistance conferred by the Nr gene. The consequences of the appearance of Nr:1 aphids for control of N. ribisnigri are discussed.

The synchrony between emergence of spring-active, insect herbivores and the budburst of their host plants could be affected by warming temperatures with influences on the availability and quality of foliage as it undergoes physical and chemical changes. This can affect the growth and survival of insects. Here, we used sun-exposed and shaded trees to determine whether the synchrony between egg hatch of western tent caterpillar, Malacosoma californicum pluviale Dyar (Lepidoptera: Lasiocampidae) and budburst of its host red alder, Alnus rubra Bongard (Betulaceae) changes with different thermal environments (temperature and light together). To explore the potential outcome of a shift in phenological synchrony, we used laboratory assays of larval growth and survival to determine the effect of variation in young, youthful and mature leaves from sun-exposed and shaded trees. While the average higher temperature of sun-exposed trees advanced the timing of budburst and egg hatch, synchrony was not disrupted. Leaf quality had no significant influence on growth or survival in the laboratory for early instars reared as family groups. Later instar larvae, however, performed best on mature leaves from sun-exposed trees. The robust relationship between leaf and larval development of western tent caterpillars suggests that warming climates may not have a strong negative impact on their success through shifts in phenological synchrony, but might influence other aspects of leaf quality and larval condition.

Parasitoids of herbivorous insects have frequently evolved specialized lineages exploiting hosts occurring on different plants. This study investigated whether host specialization is also observed when closely related parasitoids exploit herbivorous hosts sharing the same host plant. The question was addressed in economically relevant aphid parasitoids of the Lysiphlebus fabarum group. They exploit two aphid species (Aphis fabae cirsiiacanthoides and Brachycaudus cardui), co-occurring in mixed colonies (syntopy) on the spear thistle (Cirsium vulgare). Two morphologically distinguishable parasitoid lineages of the genus Lysiphlebus were observed and each showed virtually perfect host specialization on one of the two aphid species in this system. From A. f. cirsiiacanthoides, only females emerged that morphologically belonged to Lysiphlebus cardui, while males and females belonging to L. fabarum hatched from B. cardui. Microsatellite analyses indicated clear genetic differentiation of L. fabarum and L. cardui. L. cardui comprised only two distinct asexual lineages, one of which predominated throughout the area investigated. Population genetic analysis of sexual L. fabarum showed evidence for relatively strong spatial structuring and limited dispersal ability. Hyperparasitoids emerged from a large proportion of aphid mummies. One species, Pachyneuron aphidis, was significantly associated with B. cardui/L. fabarum mummies, indicating that host specialization may even extend to the trophic level above parasitoids.

The Colorado potato beetle (Leptinotarsa decemlineata, family Chrysomelidae), a phytophagous insect, which feeds preferably on potatoes, constitutes a serious pest of this crop and causes extensive damage to tomatoes and eggplants. It has a remarkable ability to develop resistance quickly against insecticides and shows a diversified and flexible life history. Consequently, the control of this pest has become difficult, requiring the development of new alternative biotechnology-based strategies. Such strategies require a thorough knowledge of the beetle's genome, including the repetitive DNA. Satellite DNA (stDNA), composed of long arrays of tandemly arranged repeat units, constitutes the major component of heterochromatin and is located mainly in centromeric and telomeric chromosomal regions. We have studied two different unrelated satellite-DNA families of which the consensus sequences were 295 and 109 bp in length, named LEDE-I and LEDE-II, respectively. Both were AT-rich (70.8% and 71.6%, respectively). Predictive models of sequence-dependent DNA bending and the study of electrophoretic mobility on non-denaturing polyacrylamide gels have shown that the DNA was curved in both satellite-DNA families. Among other features, the chromosome localization of both stDNAs has been studied. In situ hybridization performed on meiotic and mitotic nuclei showed chromosomes, including the X chromosome, with zero, one, or two stDNAs. In recent years, it has been proposed that the repetitive DNA may play a key role in biological diversification processes. This is the first molecular and cytogenetic study conducted on L. decemlineata repetitive DNA and specifically on stDNA, which is one of the important constituents of eukaryotic genomes.

Anopheles stephensi – Liston (Culicidae: Diptera) is an important urban malarial vector in the Indian sub-continent, accounting for about 15% of the total annual malaria incidence. Chemical control represents a key strategy in the management of this insect vector. However, owing to erratic and continuous application of insecticides, resistance has become a common phenomenon among them and their control has become an uphill task. The genetics of alphamethrin, a synthetic pyrethroid resistance was studied to determine its mode of inheritance. The late third instar larvae were selectively inbred for 27 and ten generations to synthesize homozygous resistant (R) and susceptible (S) stocks, respectively, to the diagnostic dose of 0.12 mg l−1. The log-dosage probit mortality relationships and degree of dominance (D) were calculated. Resistance was observed in both sexes, the dosage-mortality (d-m) line of F1 was towards the resistant parent and the ‘D’ value was found to be 0.8 indicating alphamethrin resistant (amr) gene to be autosomal and incompletely dominant. The d-m lines of F2/backcross exhibited a clear plateau of mortality across a range of doses indicating monogenic resistance. The null hypothesis for monogenic resistance was tested from mortality data of backcross progeny compared with theoretical expectations using the χ2 test and was found to be non-significant. Understanding genetics of insecticide resistance is significant in prediction and management of resistant insects. The amr genes can be used as genetic marker in A. stephensi, which can be used in several applications in conducting basic and applied genetic research.

Ribonucleic acid interference (RNAi) is a sequence-specific gene silencing mechanism induced by double-stranded RNA (dsRNA). Recently, RNAi has gained popularity as a reverse genetics tool owing to its tremendous potential in insect pest management, which includes Helicoverpa armigera. However, its efficiency is mainly governed by dsRNA concentration, frequency of application, target gene, etc. Therefore, to obtain a robust RNAi response in H. armigera, we evaluated various concentrations of dsRNA and its frequency of applications delivered through diet in silencing a midgut gene, chymotrypsin and a non-midgut gene, juvenile hormone acid methyl transferase (jhamt) of H. armigera. The extent of target gene silencing was determined by employing reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Our study revealed four significant findings: (i) single application of dsRNA elicited a delayed and transient silencing, while multiple applications resulted in early and persistent silencing of the above genes; (ii) silencing of the non-midgut gene (jhamt) through diet delivered dsRNA revealed prevalence of systemic silencing probably due to communication of silencing signals in this pest; (iii) the extent of silencing of chymotrypsin was positively correlated with dsRNA concentration and was negatively correlated with jhamt; (iv) interestingly, over-expression (15–18 folds) of an upstream gene, farnesyl diphosphate synthase (fpps), in juvenile hormone (JH) biosynthetic pathway at higher concentrations of jhamt dsRNA was the plausible reason for lesser silencing of jhamt. This study provides an insight into RNAi response of target genes, which is essential for RNAi design and implementation as a pest management strategy.

Megaplatypus mutatus (=Platypus sulcatus Chapuis) is an Ambrosia beetle native to South America, which was recently introduced in Italy and its presence there is causing severe damage to the local poplar plantations. The male M. mutatus pheromone is composed of (S)-(+)-6-methyl-5-hepten-2-ol [(+)-sulcatol], 6-methyl-5-hepten-2-one (sulcatone) and 3-pentanol. A series of field trials testing dose, blend and enantiomer composition performed in Argentina and Italy evaluated attraction and found that the optimal release rate of pheromone components as baits in cross vane baited traps (CIPEIN-CV) was 6, 6 and 30 mg day−1 of sulcatone, (+)-sulcatol and 3-pentanol, respectively. It was also determined that racemic sulcatol is as effective as the pure (+)-isomer for the purpose of beetle catch, due to the inert nature of the (−)-isomer allowing the usage of low cost racemic sulcatol instead of highly expensive (+)-sulcatol. The results of our work contribute to the development of pheromone-based local technologies with low environmental impact and low cost for control or monitoring of an important pest.

Male contributions, both chemical and behavioural, can influence female sperm usage and reproductive success. To determine whether such male factors are subject to depletion in Coccinella septempunctata and Coccinella transversalis, we tested the effects of male mating history on male virility, as estimated by measures of mating behaviour, female reproductive success and progeny fitness, with parental age held constant. Overt measures of male mating effort (wriggling duration, number of shaking bouts and total copula duration) all diminished from virgin to 5 × mated males and were mirrored by concurrent declines in female fecundity and fertility (measured over 20 days). Paternal effects were also observed which diminished as a function of mating history, suggesting that transgenerational signals of male origin are also subject to depletion. Progeny of virgin fathers had higher rates of survival (C. transversalis) and faster development (both species) than progeny of 5 × mated fathers. Seminal fluid proteins are known to have allohormonal properties and can stimulate female fecundity and fertility in a number of insects, making them strong candidates for depletion as a function of mating activity. However, it is also possible that sperm limitation and/or reduced tactile stimulation of females by multiple-mated males may have contributed to some of the observed effects.

The interplay between the host plant of an insect herbivore and an insect predator (here two-spot ladybird beetles; Adalia bipunctata (L).; Coleoptera: Coccinellidae), feeding upon such a herbivore was examined in the laboratory as factors possibly determining the differential abundance and success of green and red host races of pea aphid, Acyrthosiphon pisum Harris. The experiment comprised three treatments: two host plants (bean and clover), two treatment levels (control and predation) and three colour morph levels (green alone, red alone and green and red in mixture). Green morphs had higher fitness on the general host plant, bean Vicia faba, than on the derived host, clover (Trifolium pratense), in the absence of predation. Although green morph fitness was reduced by predation when infesting bean together with reds, there was no observable net fitness loss due to predation on clover in mixed colonies with red morphs. Red morphs exhibited fitness loss alone on both bean and clover, while clover plants seemingly prevented fitness loss in the presence of predation when red morphs were mixed with green ones. According to this scenario, when colour morphs existed as a mixed colony, the net fitness of either pea aphid morph was not influenced by predation on clover. Predators had significant effects only on red morphs on broad bean either when alone or were mixed together with green morphs. Thus, only red morphs experienced the benefits of switching from the general to the derived host red clover in the presence of predation. For green morphs, there was no apparent cost of switching host plants when they faced predation. Hence, the co-existence of green-red colour polymorphism of pea aphids on single host plants appears to be maintained by the morph gaining fitness on the derived host due to a host plant– and predation–reduction effect. These findings have important implications for understanding the ecology and evolution of host switching by different colour-plant host adapted races of pea aphids.

RNA interference (RNAi) induced through double-stranded RNA (dsRNA) has been used widely to study gene function in insects. In this paper we demonstrate the efficacy of RNAi in the cotton bollworm, Helicoverpa armigera. Using CYP6B6 as the target gene, which is expressed in the fat baby and midgut of the lepidopteran pest H. armigera, we constructed the vector which expressed dsRNA of CYP6B6. Northern blot analysis showed that dsRNA expressed in the Escherichia coli (HT115) was target gene. The results also showed that the gene expression level and protein expression level of H. armigera larvae fed with dsRNA expressed by E. coli were significantly lower than those of all controls, but the gene expression level was more obvious than that at the protein level; significant lethality differences were also found between HT115 bacteria containing L4440-dsC1 treatment and HT115 bacteria containing L4440 vector or CK (ddH2O) in instar larvae on 4 day when continuous feeding, 32.45% mortality was recorded in the group of feeding HT115 bacteria containing L4440-dsC1 on 10 day. Our results suggest that the RNAi pathway can be exploited to control insect pests.

β-Aminobutyric acid (BABA) is known to induce resistance to microbial pathogens, nematodes and insects in several host plant/pest systems. The present study was undertaken to determine whether a similar effect of BABA occurred against the Asian citrus psyllid, Diaphorina citri Kuwayama, in citrus. A 25 mM drench application of BABA significantly reduced the number of eggs/plant as compared with a water control, whereas 200 and 100 mM applications of BABA reduced the numbers of nymphs/plant and adults/plants, respectively. A 5 mM foliar application of BABA significantly reduced the number of adults but not eggs or nymphs when compared with a water control treatment. In addition, leaf-dip bioassays using various concentrations (25–500 mM) of BABA indicated no direct toxic effect on 2nd and 5th instar nymphs or adult D. citri. BABA-treated plants were characterized by significantly lower levels of iron, magnesium, phosphorus, sodium, sulfur and zinc as compared with control plants. The expression level of the PR-2 gene (β-1,3-glucanase) in BABA-treated plants that were also damaged by D. citri adult feeding was significantly higher than in plants exposed to BABA, D. citri feeding alone or control plants. Our results indicate the potential for using BABA as a systemic acquired resistance management tool for D. citri.

Rapid, precise and timely identification of invasive pest insects such as aphids is important and a challenge worldwide due to their complex life cycles, parthenogenetic reproduction, sex and colour morphs. In this respect, DNA barcoding employing a 658 bp fragment of 5′ region of the mitochondrial cytochrome oxidase I (CO-I) gene is an effective tool in addressing the above. In the present study, we employed CO-I for discriminating 142 individuals representing 32 species of aphids from India. Sequence analyses revealed that the intraspecific and interspecific distances ranged from zero to 3.8% and 2.31 to 18.9%, respectively. In addition, the study also showed for the first time the prevalence of three cryptic species, namely Brevicoryne brassicae (Linnaeus), Hyperomyzus carduellinus (Theobald) and Brachycaudus helichrysi (Kaltenbach) from India. Our work has clearly demonstrated that DNA barcoding is an efficient and accurate method for identification of aphid species (including cryptic species), an approach that potentially could play an important role in formulating viable pest management strategies, more especially biocontrol.

Following the global uptake of transgenic cotton several Hemipteran pests have emerged as primary targets for pesticide control. Previous research on one such emergent pest: the green mirid, Creontiades dilutus, indicated differential use of two crop hosts, cotton (Gossypium hirsutum, Malvaceae) and lucerne (alfalfa) (Medicago sativa, Fabaceae). We tested the hypothesis that this apparent demographic independence of lucerne and cotton inhabiting mirids is the result of cryptic species being associated with these two crops. We assessed gene flow using microsatellite markers across adjacent cotton and lucerne crops at three geographically separated sites (up to 900 km apart). We also analysed the recent feeding behaviour of these insects by amplifying chloroplast markers from their gut contents. We find high gene flow between these two crops (mean pair wise FST between host plants=0.0141 within localities), and no evidence of cryptic species. Furthermore, the gut analyses revealed evidence of substantial recent movement between these two crops. We discuss the implications of these results for interpreting multiple host use in this species and setting future research priorities for this economically important pest.