Essential Oil Trunk Injection Into Orchard Trees: Consequences on the Performance and Preference of Hemipteran Pests.

Apples and pears are among the most widely cultivated fruit species in the world. Pesticides are commonly applied using ground sprayers in conventional orchards; however, most of it will not reach the target plant, increasing the contamination of nontarget organisms such as natural predators, pollinators, and decomposers. Trunk injection is an alternative method of pesticide application that could reduce risks to beneficials and workers. Essential oils represent a 'green' alternative to pesticides due to their reported insecticidal, antimicrobial, antiviral, nematicidal, and antifungal properties. The aim of this study was, therefore, to evaluate the impact that the injection of a cinnamon essential oil solution into the trunk of apple and pear trees could have on their respective pests, Dysaphis plantaginea and Cacopsylla pyri, respectively. The feeding behavior (preference), the life history traits (performance), and the timing of this effect were measured. The injection of an essential oil emulsion in trees impacted hemipteran host-plant colonization, as for both species a modification of their preference and of their performance was observed. The feeding behavior of D. plantaginea was altered as a significantly lower proportion of aphids ingested phloem sap on injected trees, suggesting that the aphids starved to death. On the contrary, the feeding behavior of the psyllids was little changed compared to the control condition, implying that the observed mortality was due to intoxication. The results presented here could theoretically be used to control these two orchard hemipteran pests, although the effectiveness in real conditions still has to be demonstrated.

Multi-scale approach to biodiversity proxies of biological control service in European farmlands.

Intensive agriculture has profoundly altered biodiversity and trophic relationships in agricultural landscapes, leading to the deterioration of many ecosystem services such as pollination or biological control. Information on which spatio-temporal factors are simultaneously affecting crop pests and their natural enemies is required to improve conservation biological control practices. We conducted a study in 80 winter wheat crop fields distributed in three regions of North-western Europe (Brittany, Hauts-de-France and Wallonia), along intra-regional gradients of landscape complexity. Five taxa of major crop pests (aphids and slugs) and natural enemies (spiders, carabids, and parasitoids) were sampled three times a year, for two consecutive years. We analysed the influence of regional (meteorology), landscape (structure in both the years n and n-1) and local factors (hedge or grass strip field boundaries, and distance to boundary) on the abundance and species richness of crop-dwelling organisms, as proxies of the service/disservice they provide. Firstly, there was higher biocontrol potential in areas with mild winter climatic conditions. Secondly, natural enemy communities were less diverse and had lower abundances in landscapes with high crop and wooded continuities (sum of interconnected crop or wood surfaces), contrary to slugs and aphids. Finally, field boundaries with grass strips were more favourable to spiders and carabids than boundaries formed by hedges, while the opposite was found for crop pests, with the latter being less abundant towards the centre of the fields. We also revealed temporal modulation-and sometimes reversion-of the impact of local elements on crop biodiversity. To some extent, these results cause controversy because they show that hedgerows and woodlots should not be the unique cornerstones of agro-ecological landscape design strategies. We point out that combining woody and grassy habitats to take full advantage of the features and ecosystem services they both provide (biological pest control, windbreak effect, soil stabilization) may promote sustainable agricultural ecosystems. It may be possible to both reduce pest pressure and promote natural enemies by accounting for taxa-specific antagonistic responses to multi-scale environmental characteristics.

Effects of surfactins, Bacillus lipopeptides, on the behavior of an aphid and host selection by its parasitoid.

BACKGROUND: Lipopeptides extracted from the Bacillus genus are emerging biopesticides, especially in protecting crops against phytopathogens. Among the three main families of lipopeptides, surfactins have been identified as having insecticidal properties against several insect orders. However, the sublethal effects of these promising biopesticides on insect pests and their natural enemies remain largely unknown. The aim of this study was to evaluate the effects of surfactins topically applied on black bean aphid Aphis fabae mortality. First, the effects of surfactins on aphid mortality were determined by delivering increasing concentrations to adults and nymphs. Second, the sublethal effects of surfactins on locomotor activity and feeding behavior of surviving aphids were evaluated using the electropenetrography method. Finally, the effect of host exposure to surfactins on host selection behavior by Aphidius matricariae parasitoid females was analyzed. RESULTS: Four surfactins concentrations were studied (0.5, 1, 2.5 and 5 g L-1 ). There was concentration-dependent mortality in response to surfactins at 24 h after treatment. Surfactins impacted aphid behavior when delivered at 1 g L-1 by inducing a greater locomotor activity and a reduction in feeding activity. By contrast, at the third trophic level, exposure of aphid hosts to surfactins did not affect behaviors leading to host recognition and acceptance by parasitoid females. CONCLUSION: This study highlighted the consequences of aphid exposure to surfactins in the context of bottom-up regulation. Although surfactins could directly impact aphid behavior, they had no apparent consequences on the host selection behavior exhibited by parasitoid wasps.

Effects of Bacillus lipopeptides on the survival and behavior of the rosy apple aphid Dysaphis plantaginea.

Within the framework of biocontrol development, several natural lipopeptides produced by Bacillus subtilis show well-documented anti-microbial properties, especially in orchards. However, the number of studies on their putative insecticidal effects remain low despite the growing interest to develop new strategies of orchards pests' control. The rosy apple aphid Dysaphis plantaginea is the major aphid pest causing great leaf damage to apple trees. In this study, we submitted young adult aphids to topical application of three different families of lipopeptides, Plipastatin (Fengycin), Mycosubtilin (Iturin), and Surfactin, either separately or as a ternary mixture. Their aphicidal effects were investigated at 1, 2.5 and 5 g/L, both at 1 h and 24 h after exposure, and their effects on aphid behavior were studied at the 2.5 g/L concentration at 24 h after exposure. When delivered alone, lipopeptides displayed contrasted effects varying from no aphicidal activity for Mycosubtilin to a mortality induced even at low concentrations by Surfactin. Surprisingly, locomotor activity of the surviving aphids was only affected by the two least lethal treatments, Mycosubtilin and the ternary mix. Their feeding behavior was only impacted by Surfactin, the most lethal treatment, that unexpectedly increased phloem sap ingestion. The results are discussed in the context of lipopeptides applicability for integrated pest management.

Genetic structure of Malus sylvestris and potential link with preference/performance by the rosy apple aphid pest Dysaphis plantaginea.

The European crabapple Malus sylvestris, a crop wild relative of Malus domestica, is a major contributor to the cultivated apple genome and represents a potential source of interesting alleles or genes, particularly pest resistance traits. An original approach was used to explore the trophic interaction between M. sylvestris populations and its pest, the rosy apple aphid (Dysaphis plantaginea). Using 13 microsatellite markers, population genetic structure and level of crop-to-wild introgressions were inferred between M. sylvestris seedlings from three sites in Europe (Denmark, France, Romania), and M. domestica seedlings. Genetically characterized plants were also used to analyze aphid feeding behavior and fitness parameters. First, aphids submitted to two genetically close M. sylvestris populations (the Danish and French) exhibited similar behavioral parameters, suggesting similar patterns of resistance in these host plants. Second, the Romanian M. sylvestris population was most closely genetically related to M. domestica. Although the two plant genetic backgrounds were significantly differentiated, they showed comparable levels of sensitivity to D. plantaginea infestation. Third, aphid fitness parameters were not significantly impacted by the host plant's genetic background. Finally, crop-to-wild introgression seemed to significantly drive resistance to D. plantaginea independent of host plant population genetic structure, with hybrids being less suitable hosts.

Antagonist effects of the leek Allium porrum as a companion plant on aphid host plant colonization.

Combining a non-host plant (companion plant or CP) with a target cultivated plant is considered as a promising strategy to reduce pest pressure. Among the companion plants (CP) commonly used in integrated systems, those belonging to the Amaryllidaceae family (chives, garlic, onion, leek) exhibit characteristics related to certain volatile organic compounds (VOCs) with promising repellent potentialities. The aim of this work was to investigate the potential disruption of sweet pepper (host plant) colonization by the green peach aphid (Myzus persicae) when exposed to leek (Allium porrum) as a CP. Retention/dispersion, EPG and clip-cage/Petri dish laboratory experiments were thus performed to study the effect of leek VOCs on aphid settlement/migration, feeding behavior and life history traits parameters, respectively. This work revealed that leek as a CP had a negative effect on aphid feeding behavior, by disturbing the balance between phloem and xylem sap ingestion, but had no influence concerning aphid settlement. Surprisingly, leek as a CP triggered some unexpected probiotic effects on certain life history traits such as aphid survival, biomass, and fecundity, suggesting a possible hormetic effect of leek VOCs on aphid physiology. The possibility of experience-induced preference of aphids for leek VOCs was also discussed.

How the use of nitrogen fertiliser may switch plant suitability for aphids: the case of Miscanthus, a promising biomass crop, and the aphid pest Rhopalosiphum maidis.

BACKGROUND: The use of nitrogen fertiliser in agrosystems can alter plant nitrogen and consequently improve nutrient availability for herbivores, potentially leading to better performance for herbivores and higher pest pressure in the field. We compared, in laboratory conditions, the effects of nitrogen fertilisation on a promising biomass crop, Miscanthus × giganteus, and its parents M. sinensis and M. sacchariflorus. The plant-mediated effects were compared on the second trophic level, the green corn leaf aphid Rhopalosiphum maidis. RESULTS: Results showed that the biomass and leaf C:N ratio of M. sinensis plants treated with nitrogen fertiliser were significantly greater than those of non-treated plants. As regards M. × giganteus and M. sacchariflorus, the only reported change was a significantly smaller leaf C:N ratio for treated M. sacchariflorus compared with non-treated plants. Surprisingly, nitrogen fertilisation had opposite effects on plant-herbivore interactions. Following nitrogen treatments, M. sinensis was less suitable in terms of intrinsic rate of increase for R. maidis, the feeding behaviour of which was negatively affected, while M. sacchariflorus and M. × giganteus exhibited greater suitability in terms of aphid weight. CONCLUSION: Nitrogen fertilisation had contrasting effects on the three species of Miscanthus plants. These effects cascaded up to the second trophic level, R. maidis aphid pests, either through a modification of their weight or demographic parameters. The implications of these results were discussed in the context of agricultural sustainability and intensive production practices. © 2016 Society of Chemical Industry.

Aphid Parasitoid Mothers Don't Always Know Best through the Whole Host Selection Process.

Parasitoid host selection behaviour has been extensively studied in experimentally simplified tritrophic systems formed by one single food chain (one plant, one herbivore and one parasitoid species). The "Mother knows best" hypothesis predicts that the preference for a plant-host complex should be positively correlated with plant quality for offspring performance. We studied the host selection behaviour of the generalist endoparasitoid Aphidius matricariae towards the black bean aphid Aphis fabae in the intercrop system including Vicia faba as a focal plant and its companion plant Camelina sativa. Dual-choice laboratory bioassays revealed that parasitoid females preferred to orientate towards (1) the plant-aphid complex over the non-infested plant whatever the complex (2) the C. sativa-A. fabae complex over the V. faba-A. fabae complex. In dual choice attack rate bioassays, parasitoid females showed more interest towards the aphids on C. sativa but paradoxically chose to oviposit more in aphids on V. faba. Ultimately, parasitoids that had developed on the V. faba-A. fabae complex exhibited better fitness parameters. By demonstrating that parasitoid females were able to discriminate the aphid host that offered the highest fitness to their offspring but selected beforehand the least suitable plant-aphid complex, we provide key insight into the disruption in their host selection behaviour potentially triggered by diverse habitats. This suggests that the "Mother knows best" hypothesis could be thwarted by increasing the complexity of the studied systems.

Trans-generational effects of mild heat stress on the life history traits of an aphid parasitoid.

Temperature changes are common in nature and insects are particularly exposed and sensitive to such variations which can be potential stresses, ultimately affecting life history traits and overall fitness. Braconids have been widely used to study the effects of temperature on host-parasitoid interactions and the present work focused on the solitary endoparasitoid Aphidius ervi Haliday (Hymenoptera: Braconidae Aphidiidae), an efficient biological control agent commercially used against aphids such as the potato aphid Macrosiphum euphorbiae Thomas (Sternorrhyncha: Aphididae). Contrary to previous studies using heat shocks at extreme temperatures, we evaluated the effects of mild heat stresses by transferring young parasitoid adults from the constant temperature of 20°C to either a warm (25°C) or hot (28°C) temperature, for either 1 h or 48 h. Such treatments are consistent with situations commonly experienced by parasitoids when moved from their rearing conditions to greenhouses or field conditions. The effects were evaluated both on the heat stressed A. ervi adults (G0) (immediate effects) and on their first generation (G1) progeny (trans-generational effects). G0 wasps' mortality was significantly affected by the temperature in interaction with the duration of the stress. Longevity of G0 wasps surviving the heat stress was negatively affected by the temperature and females lived longer than males. Heat stress applied to A. ervi parents also had consequences on their G1 progeny whose developmental time, rates of mummification and percentage of parasitoid completing total development were negatively affected. Surprisingly, the egg load at emergence of the G1 female progeny was increased when their mothers had been submitted to a mild heat stress of 25°C or 28°C. These results clearly demonstrate trans-generational phenotypic plasticity, showing that adaptation to thermal stresses may be achieved via maternal effects. This study also sheds light on the complexity of insect responses and underlying mechanisms to fluctuating conditions in their natural environment.

Structural and functional characterization of pseudopodocyte, a shaggy immune cell produced by two Drosophila species of the obscura group.

We recently reported that most of the Drosophila species of the obscura group were unable to mount cellular capsules and no lamellocyte was ever found in the hemolymph of any of the tested species. Only three species were able to encapsulate, despite lacking lamellocytes. Their encapsulation ability was always associated with the presence of an unpreviously described kind of capsule-forming immunocytes designated as "atypical hemocytes". Here, we describe the ultrastructural and functional characteristics of this type of hemocyte. We show that these cells share many ultrastructural and morphological features with Drosophila melanogaster plasmatocytes, although they are involved in the formation of the external layers of the cellular capsule, a functional property exhibited by lamellocytes in D. melanogaster. Due to the high number of pseudopodes in these cells, we suggest to name them "pseudopodocytes". After structural and functional characterization of these atypical hemocytes, their ambiguous status between plasmatocytes and lamellocytes is discussed.

Immune resistance of Drosophila hosts against Asobara parasitoids: cellular aspects.

The immunity of Drosophila relies on a variety of defenses cooperating to fight parasites and pathogens. The encapsulation reaction is the main hemocytic response neutralizing large parasites like endophagous parasitoids. The diversity of the mechanisms of immunoevasion evolved by Asobara parasitoids, together with the wide spectrum of Drosophila host species they can parasitize, make them ideal models to study and unravel the physiological and cellular aspects of host immunity. This chapter summarizes what could be learnt on the cellular features of the encapsulation process in various Drosophila spp., and also on the major role played by Drosophila hosts hemocytes subpopulations, both in a quantitative and qualitative manner, regarding the issue of the immune Asobara-Drosophila interactions.

Strategies of avoidance of host immune defenses in Asobara species.

Eggs and larvae of endophagous parasitoids face the host's immunity reaction once they penetrate the insect host's hemocele. In order to overcome the host's immune barrier, endoparasitoids have developed various strategies. Conformer parasitoids hide and/or get protected from the attack by the host's immunity cells without interfering with the host's immune system. Differently, regulator parasitoids directly attack the host's hemocytes, therefore totally inhibiting the immunity reaction of encapsulation in the parasitized host. Female wasps may also discriminate immunoreactive hosts from nonreactive, permissive ones before laying an egg. These different strategies coexist within the same genus of the braconids Asobara, endoparasitoids of Drosophila larvae. The physiological mechanisms underlying the conformer and regulator strategies in Asobara are exposed. The factors which may contribute to the diversity of the means developed by Asobara parasitoids to overcome the hosts' immunity defenses are discussed.

Variation of success of Leptopilina boulardi in Drosophila yakuba: the mechanisms explored.

Despite an increasing knowledge of insect immune defences and virulence strategies used by parasitoids to escape them, the mechanisms underlying variation of success between parasitoid strains are still poorly understood. We have investigated this point using two lines of the parasitoid wasp Leptopilina boulardi that differ in virulence towards Drosophila yakuba. By injecting oil drops in D. yakuba larvae parasitized by virulent IS(y) females and then dissecting the larvae at different times following injection, we demonstrate that the IS(y) line alters host encapsulation ability but only during the early parasitism period. This effect is mimicked by injecting venom gland extracts, indicating that venom proteins are likely involved in immunosuppression. By contrast, the IS(m) line, unsuccessful on D. yakuba, has no immunosuppressive effect. This variation in virulence may be explained by the striking difference we report in haemocytic profiles between IS(m)- and IS(y)-parasitized larvae. We discuss our results in the light of our knowledge of the strategies evolved by Leptopilina species to counteract the D. melanogaster immune system as well as the role of parasitoid venoms in intra-specific variation of parasitoid virulence.

The fly Drosophila subobscura: a natural case of innate immunity deficiency.

The Drosophila subobscura larvae were found to be unable to form a capsule around a parasitic egg or an inert foreign body. The specificity and physiological causes of this incapacity were also explored: analysis of the circulating hemocytes showed that no lamellocyte was ever found in D. subobscura host larvae. Therefore, the fly D. subobscura is the first discovered animal species to present an innate immunodeficiency against a wide range of parasites. This is contrary to the theories that propose that all organisms, in natural conditions, are potentially able to defend themselves against parasitization. This unexpected finding opens evolutionary debates about the cost of immune resistance not only at the level of a population, but also of a whole species. We believe this species of fruitfly could become a new model system to study genes involved in hematopoïesis, and in a larger context to better understand defence reactions in organisms.

Conversely to its sibling Drosophila melanogaster, D. simulans overcomes the immunosuppressive effects of the parasitoid Asobara citri.

The endoparasitoid Asobara citri avoids Drosophila melanogaster immune defenses, thanks to immune suppressive effects. We investigated whether this parasitoid could also circumvent the immune reaction of D. simulans, a sibling species of D. melanogaster. The rates of infestation, successful parasitism, total encapsulation and mortality were measured after complete development of both D. melanogaster and D. simulans larvae parasitized by A. citri. Results showed that the parasitoids were almost never encapsulated in D. melanogaster larvae, while 45% were encapsulated in D. simulans. A. citri induced a targeted disruption of the hematopoietic organs and a decrease of the hemocytes load in host larvae of both species. Despite such disruptive immune effects most D. simulans larvae succeeded in encapsulating A. citri eggs, probably thanks to their ability to immediately mount a capsule after infestation. This work brings some insight into the diversity of the immune potentials evolved by Drosophila species towards parasitoids.

Comparative study of the strategies evolved by two parasitoids of the genus Asobara to avoid the immune response of the host, Drosophila melanogaster.

Asobara tabida and Asobara citri are two braconid endoparasitoids of Drosophila melanogaster larvae. We studied and compared the strategies evolved by these two species to avoid the immune reaction of their host. A. tabida has no negative impact on host cellular defenses and its eggs avoid encapsulation by adhering to host tissues. At the opposite, we found that A. citri, whose eggs are devoid of adhesive properties, affects the host encapsulation abilities, hemolymph phenoloxidase activity and concentrations of circulating hemocytes. Some of these effects could directly rely on a severe disruption of the hematopoietic organ anterior lobes observed in parasitized larvae. This is the first report of the immune suppressive abilities of a parasitoid from the Asobara genus. Results are presented and discussed with respect to the strategies of virulence evolved by other parasitoids to counteract the D. melanogaster immune system.