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Understanding the responses of insect herbivores to plant chemical defences is pivotal for the management of crops and pests. However, the mechanisms of interaction are not entirely understood. In this study, we compared the whole transcriptome gene expression of the aphid Macrosiphum euphorbiae grown on two different varieties of tomato that differ in their inducible chemical defences. We used two isogenic lines of tomato with a shared genetic background that only differ in the presence of type IV glandular trichomes and their associated acylsucrose excretions. This works also reports a de novo transcriptome of the aphid M. euphorbiae. Subsequently, we identified a unique and distinct gene expression profile for the first time corresponding to aphid´s exposure to type IV glandular trichomes and acylsugars. The analysis of the aphid transcriptome shows that tomato glandular trichomes and their associated secretions are highly efficient in triggering stress-related responses in the aphid, and demonstrating that their role in plant defence goes beyond the physical impediment of herbivore activity. Some of the differentially expressed genes were associated with carbohydrate, lipid and xenobiotic metabolisms, immune system, oxidative stress response and hormone biosynthesis pathways. Also, the observed responses are compatible with a starvation syndrome. The transcriptome analysis puts forward a wide range of genes involved in the synthesis and regulation of detoxification enzymes that reveal important underlying mechanisms in the interaction of the aphid with its host plant and provides a valuable genomic resource for future study of biological processes at the molecular level using this aphid.

The Old World cotton bollworm Helicoverpa armigera was first detected in Brazil with subsequent reports from Paraguay, Argentina, Bolivia, and Uruguay. This pattern suggests that the H. armigera spread across the South American continent following incursions into northern/central Brazil, however, this hypothesis has not been tested. Here we compare northern and central Brazilian H. armigera mtDNA COI haplotypes with those from southern Brazil, Uruguay, Argentina, and Paraguay. We infer spatial genetic and gene flow patterns of this dispersive pest in the agricultural landscape of South America. We show that the spatial distribution of H. armigera mtDNA haplotypes and its inferred gene flow patterns in the southwestern region of South America exhibited signatures inconsistent with a single incursion hypothesis. Simulations on spatial distribution patterns show that the detection of rare and/or the absence of dominant mtDNA haplotypes in southern H. armigera populations are inconsistent with genetic signatures observed in northern and central Brazil. Incursions of H. armigera into the New World are therefore likely to have involved independent events in northern/central Brazil, and southern Brazil/Uruguay-Argentina-Paraguay. This study demonstrates the significant biosecurity challenges facing the South American continent, and highlights alternate pathways for introductions of alien species into the New World.

Background Cassava whitefly outbreaks were initially reported in East and Central Africa cassava (Manihot esculenta Crantz) growing regions in the 1990’s and have now spread to other geographical locations, becoming a global pest severely affecting farmers and smallholder income. Whiteflies impact plant yield via feeding and vectoring cassava mosaic and brown streak viruses, making roots unsuitable for food or trading. Deployment of virus resistant varieties has had little impact on whitefly populations and therefore development of whitefly resistant varieties is also necessary as part of integrated pest management strategies. Suitable sources of whitefly resistance exist in germplasm collections that require further characterization to facilitate and assist breeding programs. Results In the present work, a hierarchical metabolomics approach has been employed to investigate the underlying biochemical mechanisms associated with whitefly resistance by comparing two naturally occurring accessions of cassava, one susceptible and one resistant to whitefly. Quantitative differences between genotypes detected at pre-infestation stages were consistently observed at each time point throughout the course of the whitefly infestation. This prevalent differential feature suggests that inherent genotypic differences override the response induced by the presence of whitefly and that they are directly linked with the phenotype observed. The most significant quantitative changes relating to whitefly susceptibility were linked to the phenylpropanoid super-pathway and its linked sub-pathways: monolignol, flavonoid and lignan biosynthesis. These findings suggest that the lignification process in the susceptible variety is less active, as the susceptible accession deposits less lignin and accumulates monolignol intermediates and derivatives thereof, differences that are maintained during the time-course of the infestation. Conclusions Resistance mechanism associated to the cassava whitefly-resistant accession ECU72 is an antixenosis strategy based on reinforcement of cell walls. Both resistant and susceptible accessions respond differently to whitefly attack at biochemical level, but the inherent metabolic differences are directly linked to the resistance phenotype rather than an induced response in the plant.

Members of the genus Torradovirus (family Secoviridae, type species Tomato torrado virus, ToTV) are spherical plant viruses transmitted by the whitefly species Trialeurodes vaporariorum and Bemisia tabaci. Knowledge on the mode of vector transmission is lacking for torradoviruses. Here, the mode of transmission was determined for Tomato marchitez virus (ToMarV). A minimal acquisition access period (AAP) and inoculation access period (IAP) of approximately 2h each was required for its transmission by T. vaporariorum, while optimal transmission required an AAP and IAP of at least 16h and 8h, respectively. Whiteflies could retain the virus under non-feeding conditions for at least 8h without loss of transmission efficiency, but upon feeding on a non-host plant in between the AAP and IAP they retained the virus for no more than 8h. Similar conditions supported transmission of isolates of ToTV and Tomato chocolàte virus (ToChV) by T. vaporariorum and B. tabaci. Additionally, similar experiments revealed the banded-winged whitefly (Trialeurodes abutilonea) as a vector for all three virus species. The results are congruent with acquisition and retention periods for semi-persistent virus transmission. RT-PCR detection analysis of ToTV and ToMarV in the vector's body revealed their presence in the stylet, but not in the head where the pharynx of the foregut is located. The results altogether indicate a semi-persistent stylet-borne mode of vector transmission for torradoviruses. Additionally, this is the first group of spherical viruses transmitted by at least three different species of whiteflies.

Solanum galapagense is closely related to the cultivated tomato and can show a very good resistance towards whitefly. A segregating population resulting from a cross between the cultivated tomato and a whitefly resistant S. galapagense was created and used for mapping whitefly resistance and related traits, which made it possible to study the genetic basis of the resistance. Quantitative trait loci (QTL) for adult survival co-localized with type IV trichome characteristics (presence, density, gland longevity and gland size). A major QTL (Wf-1) was found for adult survival and trichome characters on Chromosome 2. This QTL explained 54.1 % of the variation in adult survival and 81.5 % of the occurrence of type IV trichomes. A minor QTL (Wf-2) for adult survival and trichome characters was identified on Chromosome 9. The major QTL was confirmed in F3 populations. Comprehensive metabolomics, based on GCMS profiling, revealed that 16 metabolites segregating in the F2 mapping population were associated with Wf-1 and/or Wf-2. Analysis of the 10 most resistant and susceptible F2 genotypes by LCMS showed that several acyl sugars were present in significantly higher concentration in the whitefly resistant genotypes, suggesting a role for these components in the resistance as well. Our results show that whitefly resistance in S. galapagense seems to inherit relatively simple compared to whitefly resistance from other sources and this offers great prospects for resistance breeding as well as elucidating the underlying molecular mechanism(s) of the resistance.

The two-spotted spider mite, Tetranychus urticae, is a chelicerate herbivore with an extremely wide host range and an extraordinary ability to develop pesticide resistance. Due to its responsiveness to natural and synthetic xenobiotics, the spider mite is becoming a prime pest herbivore model for studies of the evolution of host range, plant-herbivore interactions and mechanisms of xenobiotic resistance. The spider mite genome has been sequenced and its transcriptional responses to developmental and various biotic and abiotic cues have been documented. However, to identify biological and evolutionary roles of T. urticae genes and proteins, it is necessary to develop methods for the efficient manipulation of mite gene function or protein activity. Here, we describe protocols developed for the delivery of small molecules into spider mites. Starting with mite maintenance and the preparation of the experimental mite populations of developmentally synchronized larvae and adults, we describe 3 methods for delivery of small molecules including artificial diet, leaf coating, and soaking. The presented results define critical steps in these methods and demonstrate that they can successfully deliver tracer dyes into mites. Described protocols provide guidelines for high-throughput setups for delivery of experimental compounds that could be used in reverse genetics platforms to modulate gene expression or protein activity, or for screens focused on discovery of new molecules for mite control. In addition, described protocols could be adapted for other Tetranychidae and related species of economic importance such as Varroa, dust and poultry mites., Correction -- 15 Dec 2017: Suzuki T, España MU, Nunes MA, Zhurov V, Dermauw W, et al. (2017) Correction: Protocols for the delivery of small molecules to the two-spotted spider mite, Tetranychus urticae. PLOS ONE 12(12): e0190025.

International audience; Wheat being staple food of Pakistan is constantly attacked by major wheat aphid species, Schizaphis graminum (R.), Rhopalosiphum padi (L.) and Sitobion avenae (F.). Due to concern on synthetic chemical use in wheat, it is imperative to search for alternative environment-and human-friendly control measures such as botanical pesticides. In the present study, we evaluated the comparative role of neem seed extract (NSE), moringa leaf extract (MLE) and imidacloprid (I) in the management of the aphid as well as the yield losses parameters in late planted wheat fields. Imidacloprid reduced significantly aphids infestation compared to the other treatments, hence resulting in higher yield, particularly when applied with MLE. The percentages of yield increase in I+MLE treated plots over the control were 19.15-81.89% for grains per spike, 5.33-37.62% for thousand grain weight and 27.59-61.12% for yield kg/ha. NSE was the second most effective control measure in suppressing aphid population, but the yield protected by NSE treatment over the control was comparable to that by imidacloprid. Population densities of coccinellids and syrphids in the plots treated with NSE-2 were higher than those treated with imidacloprid in two out of three experiments during 2013-14. Low predator density in imidacloprid-treated plots was attributed to the lower availability of prey aphids. The efficacy of NSE against aphids varied depending on degree of synchronization among the application timing, the activity of aphids, crop variety and environmental conditions. Despite that, we suggested NSE to be a promising alternative botanical insecticide compared to the most commonly recommended imidiacloprid. Further studies should consider the side effects of biopesticides on non-target organisms in order to provide better management practices in the field.

In China, two invasive pests, Bemisia tabaci MEAM1 (Gennadius) and Frankliniella occidentalis (Pergande), often co-occur with the native pest, Aphis gossypii (Glover), on plants of Malvaceae and Cucurbitaceae. All three are preyed on by the native ladybird, Harmonia axyridis (Pallas); however, the native predator might be expected to prefer native prey to the exotic ones due to a shared evolutionary past. In order to clarify whether the presence of native prey affected the consumption of these two invasive species by the native predator, field-cage experiments were conducted. A duplex qPCR was used to simultaneously detect both non-native pests within the gut of the predator. H. axyridis readily accepted both invasive prey species, but preferred B. tabaci. With all three prey species available, H. axyridis consumption of B. tabaci was 39.3 +/- 2.2% greater than consumption of F. occidentalis. The presence of A. gossypii reduced (by 59.9% on B. tabaci, and by 60.6% on F. occidentalis), but did not stop predation on the two exotic prey when all three were present. The consumption of B. tabaci was similar whether it was alone or together with A. gossypii. However, the presence of aphids reduced predation on the invasive thrips. Thus, some invasive prey may be incorporated into the prey range of a native generalist predator even in the presence of preferred native prey.

The cabbage whitefly [Aleyrodes proletella L. (Hemiptera: Aleyrodidae)] is becoming a serious pest in Brassica oleracea L. (Brassicaceae) crops. However, almost nothing is known about the interaction of this insect with its host plants. Previous studies have shown differences in the natural occurrence of adults, eggs, and nymphs on the closely related B. oleracea cultivars Christmas Drumhead and Riviera grown in the field. In this study, we aimed to identify the nature of these differences and to gain insight into the resistance mechanisms against A. proletella. We used no-choice experiments on field- and greenhouse-grown plants to show that the differences between the two cultivars are mainly based on antibiosis (traits that reduce herbivore performance) and not on antixenosis (traits that deter herbivory). This was further supported by laboratory choice experiments that indicated little or no discrimination between the two cultivars based on plant volatiles. We showed that resistance is dependent on plant age, that is, resistance increased during plant development, and is mainly independent of environmental factors. Analysis of probing behaviour revealed that the resistance trait affects A. proletella at the phloem level and that morphological differences between the two cultivars are most likely not involved. We suggest that compounds present in the phloem reduce sap ingestion by the whitefly and that this explains the observed resistance

Biological control of different species of pest with various species of generalist predators can potentially disrupt the control of pests through predator-predator interactions. We evaluate the impact of three species of generalist predatory mites on the biological control of green peach aphids, Myzus persicae (Sulzer) with the aphidophagous gall midge Aphidoletes aphidimyza (Rondani). The predatory mites tested were Neoseiulus cucumeris (Oudemans), Iphiseius degenerans (Berlese) and Amblyseius swirskii Athias-Henriot, which are all commonly used for pest control in greenhouse sweet pepper. All three species of predatory mites were found to feed on eggs of A. aphidimyza, even in the presence of abundant sweet pepper pollen, an alternative food source for the predatory mites. In a greenhouse experiment on sweet pepper, all three predators significantly reduced population densities of A. aphidimyza, but aphid densities only increased significantly in the presence of A. swirskii when compared to the treatment with A. aphidimyza only. This stronger effect of A. swirskii can be explained by the higher population densities that this predator reached on sweet pepper plants compared to the other two predator species. An additional experiment showed that female predatory midges do not avoid oviposition sites with the predator A. swirskii. On the contrary, they even deposited more eggs on plants with predatory mites than on plants without. Hence, this study shows that disruption of aphid control by predatory mites is a realistic scenario in sweet pepper, and needs to be considered when optimizing biological control strategies.

Tomato yellow leaf curl disease, a devastating disease of Solanum lycopersicum (tomato), is caused by a complex of begomoviruses generally referred to as Tomato yellow leaf curl virus (TYLCV). Almost all breeding for TYLCV resistance has been based on the introgression of the Ty-1 resistance locus derived from Solanum chilense LA1969. Knowledge about the exact location of Ty-1 on tomato chromosome 6 will help in understanding the genomic organization of the Ty-1 locus. In this study, we analyze the chromosomal rearrangement and recombination behavior of the chromosomal region where Ty-1 is introgressed. Nineteen markers on tomato chromosome 6 were used in F2 populations obtained from two commercial hybrids, and showed the presence of a large introgression in both. Fluorescence in situ hybridization (FISH) analysis revealed two chromosomal rearrangements between S. lycopersicum and S. chilense LA1969 in the Ty-1 introgression. Furthermore, a large-scale recombinant screening in the two F2 populations was performed, and 30 recombinants in the Ty-1 introgression were identified. All recombination events were located on the long arm beyond the inversions, showing that recombination in the inverted region was absent. Disease tests on progenies of informative recombinants with TYLCV mapped Ty-1 to the long arm between markers MSc05732-4 and MSc05732-14, an interval overlapping with the reported Ty-3 region, which led to the indication that Ty-1 and Ty-3 may be allelic. With this study we prove that FISH can be used as a diagnostic tool to aid in the accurate mapping of genes that were introgressed from wild species into cultivated tomato.

A monoclonal antibody (McAb), EGA-4A9, was developed to detect the semi-digested proteins of the English grain aphid, Sitobion avenae (Fabricius) (Hemiptera: Aphididae), in predatory ladybird beetles (species of the genera Adonia, Coccinella, Hippodamia, and Propylea) using the gut homogenate of Adonia variegata (Goeze) (Coleoptera: Coccinellidae) adults which had fed on S. avenae as immunogen. The McAb was selected by screening hybridoma lines for antibodies that bound with the semi-digested aphid proteins in ladybirds. A double antibody sandwich enzyme-linked immuno-sorbent assay (ELISA) using ClonotypingTM System/HRP showed that it belonged to the IgG2a isotype. It did not cross-react with any of the 21 arthropod species tested besides the ladybird beetles fed on S. avenae with an indirect ELISA. It could still detect the semi-digested proteins in the gut of a ladybird adult, kept at 25 °C, that had ingested one aphid 6 days before. The extended antigen detection period and the high specificity of the antibody indicated that EGA-4A9 could be used to study interactions between English grain aphids and their ladybird predators in the field. Between 28 and 72% of coccinellids collected from the field were positive for English grain aphid protein by ELISA. The percentage of McAb-positive predatory ladybird beetles was positively correlated with the density of S. avenae in wheat fields

We studied the capacity of one species of predator to control two major pests of greenhouse crops, Western flower thrips (Frankliniella occidentalis (Pergande)) and the greenhouse whitefly (Trialeurodes vaporariorum (Westwood)). In such a one-predator¿two-prey system, indirect interactions can occur between the two pest species, such as apparent competition and apparent mutualism. Whereas apparent competition is desired because it brings pest levels down, apparent mutualism is not, because it does the opposite. Because apparent competition and apparent mutualism occurs at different time scales, it is important to investigate the effects of a shared natural enemy on biological control on a time scale relevant for crop growth. We evaluated the control efficacy of the predatory mites Amblyseius swirskii (Athias-Henriot) and Euseius ovalis (Evans) in cucumber crops in greenhouse compartments with only thrips, only whiteflies or both herbivorous insects together. Each of the two predators controlled thrips, but A. swirskii reduced thrips densities the most. There was no effect of the presence of whiteflies on thrips densities. Whitefly control by each of the two predators in absence of thrips was not sufficient, yet better with E. ovalis. However, whitefly densities in presence of thrips were reduced dramatically, especially by A. swirskii. The densities of predators were up to 15 times higher in presence of both pests than in the single-pest treatments. Laboratory experiments with A. swirskii suggest that this is due to a higher juvenile survival and developmental rate on a mixed diet. Hence, better control may be achieved not only because of apparent competition, but also through a positive effect of mixed diets on predator population growth. This latter phenomenon deserves more attention in experimental and theoretical work on biological control and apparent competition.

Planting Bt cotton in China since 1997 has led to important changes in the natural enemy communities occurring in cotton, however their specific effect on suppressing the cotton aphids (being notorious in conventional cotton ecosystem) has not been fully documented yet. We observed strong evidence for top-down control of the aphid population, e.g. the control efficiency of natural enemies on cotton aphid increased significantly in open field cages compared to exclusion cages, accounted for 60.2, 87.2 and 76.7% in 2011, 2012 and 2013 season, respectively. The cotton aphid populations peaked in early June to late July (early and middle growth stages) in open field cotton survey from 2011 to 2013. The population densities of cotton aphids and natural enemies were highest on middle growth stage while lowest densities were recorded on late stage for aphids and on early plant stage for natural enemies. Aphid parasitoids (Trioxys spp., Aphidius gifuensis), coccinellids and spiders were key natural enemies of cotton aphid. Briefly, natural enemies can suppress aphid population increase from early to middle plant growth stages by providing biocontrol services in Chinese Bt cotton.

Plant responses induced by zoophytophagous plant bugs (Miridae) have not been thoroughly studied. Here, we show that three different zoophytophagous predators (Nesidiocoris tenuis, Macrolophus pygmaeus and Dicyphus maroccanus) have different capacities for the induction of responses in tomato plants, resulting in varying degrees of attractiveness of the plants to pests and natural enemies. Tomato plants punctured by N. tenuis were less attractive to the whitefly Bemisia tabaci and to the lepidopteran Tuta absoluta. In contrast, tomato plants punctured by M. pygmaeus and D. maroccanus were not able to repel B. tabaci and, more interestingly, became more attractive to T. absoluta. The ability of N. tenuis to make tomato plants less attractive to B. tabaci was attributed to the activation of the abscisic acid (ABA) signaling pathway, which was only up-regulated in plants punctured by N. tenuis. However, the phytohormones involved in the behavioral responses of T. absoluta could not be identified; therefore, further studies are required. Additionally, all three zoophytophagous mirid predators activated jasmonic acid (JA) signaling pathways, which resulted in the parasitoid Encarsia formosa being attracted to tomato plants. Here, the implications of these results on the efficacy of these three predators as biocontrol agents are discussed. The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under the Grant agreement n°265865 and by the Spanish Ministry of Science and Innovation (AGL2011-30538-C03) and the Conselleria d’Agricultura, Pesca i Alimentació de la Generalitat Valenciana. The authors thank Javier Calvo (KOPPERT BS) for supplying the insects and Universitat Jaume I-SCIC for providing technical support.

International audience; Phytoviruses are highly prevalent in plants worldwide, including vegetables and fruits. Humans, and more generally animals, are exposed daily to these viruses, among which several are extremely stable. It is currently accepted that a strict separation exists between plant and vertebrate viruses regarding their host range and pathogenicity, and plant viruses are believed to infect only plants. Accordingly, plant viruses are not considered to present potential pathogenicity to humans and other vertebrates. Notwithstanding these beliefs, there are many examples where phytoviruses circulate and propagate in insect vectors. Several issues are raised here that question if plant viruses might further cross the kingdom barrier to cause diseases in humans. Indeed, there is close relatedness between some plant and animal viruses, and almost identical gene repertoires. Moreover, plant viruses can be detected in non-human mammals and humans samples, and there are evidence of immune responses to plant viruses in invertebrates, non-human vertebrates and humans, and of the entry of plant viruses or their genomes into non-human mammal cells and bodies after experimental exposure. Overall, the question raised here is unresolved, and several data prompt the additional extensive study of the interactions between phytoviruses and non-human mammals and humans, and the potential of these viruses to cause diseases in humans.

Bemisia tabaci (Gennadius) is a serious pest of vegetable, ornamental, and agronomic crops throughout the world. To control B. tabaci, Eretmocerus eremicus Rose & Zolnerowich, and Eretmocerus mundus Mercet are considered the most effective parasitoids in dry tropical regions. In parasitoids, choosing the `right` hosts has direct consequences for their reproductive success and efficiency as biocontrol agent. Therefore, being able to discriminate a parasitized host from an unparasitized one would be important to prevent wasting time, eggs, and to reduce the mortality risk for their offspring. We evaluated intra- and interspecific host discrimination and the chance of super-parasitism or multi-parasitism in two populations of E. mundus (sexual and asexual) and E. eremicus. Different combinations and sequences of female introduction were carried out for the various populations and species. Experienced females avoided super-parasitism. However, naïve females did lay eggs under hosts that were previously parasitized by conspecific females. E. eremicus females avoided to multi-parasitize hosts parasitized by E. mundus. However, E. mundus females did multi-parasitize the hosts that had been parasitized earlier by E. eremicus. In the case of super-parasitism, the outcome showed that neither of the E. mundus populations was stronger, whereas in the case of multi-parasitism E. mundus appeared stronger than E. eremicus. Since those populations and species are morphologically similar a molecular method had to be developed to identify the outcome of super- or multi-parasitism, which is presented in Appendix A.

Insect parasitoids lay their eggs in arthropods. Some parasitoid species not only use their arthropod host for oviposition but also for feeding. Host feeding provides nutrients to the adult female parasitoid. However, in many species, host feeding destroys an opportunity to oviposit. For parasitoids that attack Homoptera, honeydew is a nutrient-rich alternative that can be directly imbibed from the host anus without injuring the host. A recent study showed that feeding on host-derived honeydew can be an advantageous alternative in terms of egg quantity and longevity. Here we explore the conditions under which destructive host feeding can provide an advantage over feeding on honeydew. For 5 days, Encarsia formosa Gahan (Hymenoptera: Aphelinidae) parasitoids were allowed daily up to 3 h to oviposit until host feeding was attempted. Host feedings were either prevented or allowed and parasitoids had ad libitum access to honeydew between foraging bouts. Even in the presence of honeydew, parasitoids allowed to host feed laid more eggs per hour of foraging per host-feeding attempt than parasitoids that were prevented from host feeding. The higher egg-laying rate was not compromised by survival or by change in egg volume over time. In conclusion, host feeding can provide an advantage over feeding on honeydew. This applies most likely under conditions of high host density or low extrinsic mortality of adult parasitoids, when alternative food sources cannot supply enough nutrients to prevent egg limitation. We discuss how to integrate ecological and physiological studies on host-feeding behavior.

Semi-natural habitats in agricultural landscapes may support parasitoid populations that provide biocontrol services by suppressing populations of crop pests, but little is known about the spatial pattern and variability of these services at different levels of scale. Here we investigate the rarely studied phenomenon of early-season crop colonization by parasitoids and the relationship with the surrounding landscape. We assessed parasitism of whiteflies by placing whitefly infested cotton seedlings in remnant vegetation, arable land 25–125 m from remnant vegetation, and arable land further than 400 m from remnant vegetation. Twelve to twenty sentinel plants were exposed in a 25 × 25 m grid pattern in plots in each habitat. The experiment was conducted at 18 locations across two landscapes and repeated three times in a 2-week period in 2007 and 2008. Parasitism was observed during the first three days after the introduction of the whitefly infested seedlings and was in all cases caused by Encarsia spp. The mean number of parasitized whitefly per plant was 0.106 ± 0.025 and was highest on cotton plants placed in remnant vegetation, declining with increasing distance from remnant vegetation. A regression model with land use and meteorological variables received more statistical support from the data than models with only landscape and time period as factors. Parasitism levels were influenced by the proportion of remnant vegetation, grassland, as well as wind, temperature, dew point temperature and year. Early-season colonization of whitefly infested seedlings by parasitoids was erratic and characterized by large spatial (inter-plant and inter-plot) and temporal variation. Our study confirms that remnant vegetation function as reservoirs for parasitoids and that parasitoids can penetrate arable fields beyond 125 m within 3 days. However, variation in the occurrence of parasitism makes it difficult to predict parasitoid colonization at a specific place and time. Therefore, field-based scouting for pests and parasitoids is necessary, even in landscapes with a high biocontrol potential.

Bead-based suspension array systems enable simultaneous fluorescence-based identification of multiple nucleic acid targets in a single reaction. This study describes the development of a novel approach to plant virus and vector diagnostics, a multiplexed 7-plex array that comprises a hierarchical set of assays for the simultaneous detection of begomoviruses and Bemisia tabaci, from both plant and whitefly samples. The multiplexed array incorporates genus, species and strain-specific assays, offering a unique approach for identifying both known and unknown viruses and B. tabaci species. When tested against a large panel of sequence-characterized begomovirus and whitefly samples, the array was shown to be 100% specific to the homologous target. Additionally, the multiplexed array was highly sensitive, efficiently and concurrently determining both virus and whitefly identity from single viruliferous whitefly samples. The detection limit for one assay within the multiplexed array that specifically detects Tomato yellow leaf curl virus-Israel (TYLCV-IL) was quantified as 200 fg of TYLCV-IL DNA, directly equivalent to that of TYLCVspecific qPCR. Highly reproducible results were obtained over multiple tests. The flexible multiplexed array described in this study has great potential for use in plant quarantine, biosecurity and disease management programs worldwide. (C) 2014 Elsevier B.V. All rights reserved.

Long-term effects of a single application of imidacloprid on ladybird beetle, Coccinella septempunctata L., were studied in indoor laboratory microcosms, starting with the 2nd instar larvae of C. septempunctata but covering the full life cycle. The microcosms comprised enclosures containing a pot with soil planted with broad bean plants and black bean aphid, Aphis craccivora Koch, as food. Exposure doses (0.85-13.66g a.i. ha(-1)) in the long-term microcosm experiment were based on a preliminary short-term (72h) toxicity test with 2nd instar larvae. The measurement endpoints used to calculate NOERs (No Observed Effect application Rates) included development time, hatching, pupation, adult emergence, survival and number of eggs produced. Furthermore, for these endpoints ER50 (application rate causing 50 percent effect) and LR50 (application rate causing 50 percent mortality) values were calculated when possible. The single imidacloprid application affected survival (lowest LR50 4.07g a.i. ha(-1); NOER 3.42g a.i. ha(-1)), egg production (ER50 26.63g a.i. ha(-1)) and egg hatching (NOER 6.83g a.i. ha(-1)). Statistically significant treatment-related effects on the whole development duration, pupation and adult emergence could not be demonstrated (NOER≥13.66g a.i. ha(-1)). The lowest L(E)R50 values and NOERs derived from the laboratory microcosm test with C. septempunctata are lower than the reported field application rates of imidacloprid (15-60g a.i. ha(-1)) in cotton cultivation in China, suggesting potential risks to beneficial arthropods.

Eriopis connexa is a native coccinelid predator in the Neotropical Region. In Argentina it is commonly found associated to sucking pests in several crops and among them aphids and whiteflies. These pests are usually controlled with newly developed systemic insecticides, such as the neonicotinoids. However, the compatibility between selective pesticides and natural enemies is required before incorporating them in integrated pest management (IPM) packages. Within this frame, the objective of this study was to evaluate the side effect of various concentrations/doses of one commonly used neonicotinoid in vegetal crops, acetamiprid, on immature stages of E. connexa by dipping or topical exposure for eggs and larvae, respectively. Acetamiprid reduced egg hatching from 34 to 100 %. Moreover, the embryogenesis was disrupted by insecticide at early embryo stage at all tested concentrations. Second larval instar was more susceptible to acetamiprid than the fourth one and this susceptibility was positively related with the tested concentrations. On the other hand, the survival reduction at larval stage reached 100 % from 20 mg a.i./L (10 % of maximum field concentration). Besides, the reproduction of the females developed from topical bioassays on fourth instar larvae was strongly affected, with reduction in fecundity and fertility from 22 to 44 % and from 37 to 45 %, respectively. Overall the results showed a high toxicity of acetamiprid on immature stages of E. connexa, demonstrating that this broadly used insecticide could reduce biocontrol services provided by this predator and could also likely disturb IPM programs., Centro de Investigaciones del Medioambiente, Centro de Estudios Parasitológicos y de Vectores

The cabbage whitefly [Aleyrodes proletella L. (Hemiptera: Aleyrodidae)] is becoming a serious pest in Brassica oleracea L. (Brassicaceae) crops. However, almost nothing is known about the interaction of this insect with its host plants. Previous studies have shown differences in the natural occurrence of adults, eggs, and nymphs on the closely related B. oleracea cultivars Christmas Drumhead and Riviera grown in the field. In this study, we aimed to identify the nature of these differences and to gain insight into the resistance mechanisms against A. proletella. We used no-choice experiments on field- and greenhouse-grown plants to show that the differences between the two cultivars are mainly based on antibiosis (traits that reduce herbivore performance) and not on antixenosis (traits that deter herbivory). This was further supported by laboratory choice experiments that indicated little or no discrimination between the two cultivars based on plant volatiles. We showed that resistance is dependent on plant age, that is, resistance increased during plant development, and is mainly independent of environmental factors. Analysis of probing behaviour revealed that the resistance trait affects A. proletella at the phloem level and that morphological differences between the two cultivars are most likely not involved. We suggest that compounds present in the phloem reduce sap ingestion by the whitefly and that this explains the observed resistance

Tomato yellow leaf curl disease, a devastating disease of Solanum lycopersicum (tomato), is caused by a complex of begomoviruses generally referred to as Tomato yellow leaf curl virus (TYLCV). Almost all breeding for TYLCV resistance has been based on the introgression of the Ty-1 resistance locus derived from Solanum chilense LA1969. Knowledge about the exact location of Ty-1 on tomato chromosome 6 will help in understanding the genomic organization of the Ty-1 locus. In this study, we analyze the chromosomal rearrangement and recombination behavior of the chromosomal region where Ty-1 is introgressed. Nineteen markers on tomato chromosome 6 were used in F2 populations obtained from two commercial hybrids, and showed the presence of a large introgression in both. Fluorescence in situ hybridization (FISH) analysis revealed two chromosomal rearrangements between S. lycopersicum and S. chilense LA1969 in the Ty-1 introgression. Furthermore, a large-scale recombinant screening in the two F2 populations was performed, and 30 recombinants in the Ty-1 introgression were identified. All recombination events were located on the long arm beyond the inversions, showing that recombination in the inverted region was absent. Disease tests on progenies of informative recombinants with TYLCV mapped Ty-1 to the long arm between markers MSc05732-4 and MSc05732-14, an interval overlapping with the reported Ty-3 region, which led to the indication that Ty-1 and Ty-3 may be allelic. With this study we prove that FISH can be used as a diagnostic tool to aid in the accurate mapping of genes that were introgressed from wild species into cultivated tomato.

The solitary parasitoid Eretmocerus mundus Mercet (Hymenoptera: Aphelinidae) is one of the key biological control agents of the whitefly Bemisia argentifolii Bellows & Perring, also known as B. tabaci (Gennadius) B-biotype (Homoptera: Aleyrodidae). We present new information on its biology, and particularly about larval elimination during the first instar in cases of superparasitism. For the first time, physical elimination of supernumerary larvae was observed, both outside and inside the host. These findings are documented with confocal microscopy images and video recordings. We observed more than 350 B. argentifolii nymphs, parasitized with one, two or more than two larvae. Physical attack took place only when parasitoid eggs were laid in contact with each other. In this case the percentage larval mortality outside the host was significantly higher (45.7%) than in cases of single larvae (12.0%) or larvae hatched under the same host but not in contact with each other (23.3%). These new findings are important in view of a possible horizontal transfer of a parthenogenesis inducing Wolbachia bacterium from a thelytokous to an arrhenotokous strain, with the aim of augmenting the biological control of B. argentifolii.

Eretmocerus species (Hym. Aphelinidae) are solitary parasitoids of Bemisia tabaci (Gennadius). Mate finding and mating behavior of two species, E. mundus and E. eremicus, were studied under laboratory conditions. We used three populations of Eretmocerus: typical arrhenotokous populations of E. eremicus ( from USA) and E. mundus ( from Spain), and an atypical thelytokous population of E. mundus ( from Australia). We studied the intra- and interspecific responses of males to volatile and nonvolatile components of the female sex pheromones, mating behavior, and hybridization between populations and species. In both arrhenotokous populations, males reacted to volatile pheromones by walking toward conspecific virgin females. Males also reacted to nonvolatile pheromones by spending more time on and around patches on leaves of poinsettia plants that had been exposed to virgin females. Males of E. eremicus showed the same reaction to the nonvolatile sex pheromone of E. mundus females, but E. mundus males did not show any reaction to the nonvolatile sex pheromone of E. eremicus. There was no response of males of both species to thelytokous females of E. mundus. In both species three phases were distinguished in the mating behavior: premating, mating, and postmating. The duration of the phases differed between the three populations. Successful copulation between the two Eretmocerus species did not occur. In contrast, we recorded some successful copulations between Australian males and Spanish females of E. mundus, but they did not produce any hybrid females.

International audience; A new icosahedral DNA virus was isolated from aphids (Myzus persicae) that showed abnormal growth and development. The purified virus particles have a diameter of 20 nm and contain a single-stranded DNA molecule of approximately 5.7 kb. The viral particles are composed of five structural proteins (92, 85, 68, 64, and 57 kDa). As the main biophysical properties of this virus are similar to those of the members of the genus Densovirus it was tentatively named Myzus persicae densovirus (MpDNV). A PCR-based detection method and a polyclonal antiserum raised against MpDNV allowed the detection of the virus in a single-infected aphid. MpDNV is immunologically related to Junonia coenia densovirus, but not to other members of the subfamily Densovirinae. Biological assays showed that MpDNV could be both transmitted transovarially and horizontally via honeydew and saliva. MpDNV was able to infect whiteflies but not other aphid species tested.

International audience; The genomes of two Watermelon chlorotic stunt virus (WmCSV) isolates, one from the Sudan and one from Iran, were cloned and sequenced. Sequence relationship with other geminiviruses characterizes WmCSV as a typical Eastern Hemisphere geminivirus with a bipartite genome. The two geographically distant WmCSV isolates from Africa and the Middle East share a very high overall sequence similarity: 98% between their DNA-A and 96% between their DNA-B components, and their respective capsid proteins are identical. A single amino acid change in the capsid protein (N131D) renders WmCSV whitefly nontransmissible. This region of the capsid is also implicated in transmission by Bemisia tabaci of Tomato yellow leaf curl virus.

Background Phloem-feeding insects are among the most devastating pests worldwide. They not only cause damage by feeding from the phloem, thereby depleting the plant from photo-assimilates, but also by vectoring viruses. Until now, the main way to prevent such problems is the frequent use of insecticides. Applying resistant varieties would be a more environmental friendly and sustainable solution. For this, resistant sources need to be identified first. Up to now there were no methods suitable for high throughput phenotyping of plant germplasm to identify sources of resistance towards phloem-feeding insects. Results In this paper we present a high throughput screening system to identify plants with an increased resistance against aphids. Its versatility is demonstrated using an Arabidopsis thaliana activation tag mutant line collection. This system consists of the green peach aphid Myzus persicae (Sulzer) and the circulative virus Turnip yellows virus (TuYV). In an initial screening, with one plant representing one mutant line, 13 virus-free mutant lines were identified by ELISA. Using seeds produced from these lines, the putative candidates were re-evaluated and characterized, resulting in nine lines with increased resistance towards the aphid. Conclusions This M. persicae-TuYV screening system is an efficient, reliable and quick procedure to identify among thousands of mutated lines those resistant to aphids. In our study, nine mutant lines with increased resistance against the aphid were selected among 5160 mutant lines in just 5 months by one person. The system can be extended to other phloem-feeding insects and circulative viruses to identify insect resistant sources from several collections, including for example genebanks and artificially prepared mutant collections.