Effect of the Landscape on Insect Pests and Associated Natural Enemies in Greenhouses Crops: The Strawberry Study Case.

Compared to open-field crops, the influence of the surrounding landscape on insect diversity in greenhouse crops has been poorly studied. Due to growing evidence of insect influx in greenhouses, identifying the landscape properties influencing the protected crop colonization by insect pests and their natural enemies would promote the improvement of both pest prevention and conservation biological control methods. Here, we present a field study on the effect of the surrounding landscape on the colonization of greenhouse crops by insect pests and associated natural enemies. By monitoring 32 greenhouse strawberry crops in the South West of France, we surveyed crop colonization by four insect pests and four natural enemy groups over two cultivation periods. Our results showed that the landscape structure and composition could have contrasting effects on insect colonization of greenhouse crops so there could be species-specific effects and not general ones. While the degree of openness of greenhouses and the pest management practices modulated insect diversity marginally, we also showed that seasonality represented a key factor in insect crop colonization. The various responses of insect pests and natural enemy groups to the landscape support the idea that pest management methods must involve the surrounding environment.

Genetics of wild and mass-reared populations of a generalist aphid parasitoid and improvement of biological control.

Due to their ability to parasitize various insect species, generalist parasitoids are widely used as biological control agents. They can be mass-reared and released in agroecosystems to control several pest species in various crops. However, the existence of genetic differentiation among populations of generalist parasitoid species is increasingly recognized and this can be associated with an adaptation to local conditions or to a reduced range of host species. Moreover, constraints of mass-rearing conditions can alter genetic variation within parasitoid populations released. These features could be associated with a reduced efficiency of the control of targeted pest species. Here, we focused on strawberry greenhouses where the control of aphids with the generalist parasitoid Aphidius ervi appears to be inefficient. We investigated whether this inefficiency may have both genetic and ecological bases comparing wild and commercial populations of A. ervi. We used two complementary genetic approaches: one based on the mitochondrial marker COI and one based on microsatellite markers. COI analysis showed a genetic differentiation within the A. ervi species, but the structure was neither associated with the commercial/wild status nor with host species factors. On the other hand, using microsatellite markers, we showed a genetic differentiation between commercial and wild A. ervi populations associated with a loss of genetic diversity within the mass-reared populations. Our ecological genetics study may potentially explain the weak efficiency of biological control of aphids in protected strawberry crops and enable to provide some insights to improve biological control.

"Generalist" Aphid Parasitoids Behave as Specialists at the Agroecosystem Scale.

The degree of trophic specialization of interacting organisms impacts on the structure of ecological networks and has consequences for the regulation of crop pests. However, it remains difficult to assess in the case of parasitoids. Host ranges are often established by listing host records from various years and geographic areas in the literature. Here, we compared the actual hosts exploited at a local farm-scale by aphid parasitoids (Hymenoptera: Aphidiinae), to the available species listed as hosts for each parasitoid species. We sampled aphids and their parasitoids in cultivated and uncultivated areas in an experimental farm from April to November 2014 and thereafter used DNA-based data to determine whether a differentiation in sequences existed. Twenty-nine parasitoid species were found on 47 potential aphid hosts. Our results showed that the great majority of the parasitoid tested used fewer host species than expected according to data published in the literature and parasitized a limited number of hosts even when other potential hosts were available in the environment. Moreover, individuals of the most generalist species differed in their DNA sequences, according to the aphid species and/or the host plant species. At a local scale, only obligate or facultative specialist aphid parasitoids were detected. Local specialization has to be considered when implementing the use of such parasitoids in pest regulation within agroecosystems.

Larval hitch-hiking and adult flight are two ways of aphidiinae parasitoids long-range dispersal.

Dispersal strategies and success of pests' natural enemies widely influence the efficiency of biological control. In this study, we compare two dispersal strategies among Aphidiinae parasitoids: eggs and larvae dispersal through winged aphid flight and active dispersal by adult parasitoids. Using a molecular method applied to a sample of >2,000 winged migratory aphids captured in a suction trap situated in Western France, we assessed the proportion of winged aphids carrying an aphidiine larva. In the six most abundant aphid species, we found an average parasitism rate of migrating aphids close to 1% and identified seven different, mainly generalist, parasitoid species. We also identified the species and the sex of adult Aphidiinae captured by the suction trap based on morphological criteria. We found that dispersing adult parasitoids were almost exclusively female. Parasitoid dispersal strategy seems to be species-dependant but this result needs to be confirmed by an exhaustive analysis of winged aphids captured. We discuss the possible impact of the low parasitism rate of winged aphids on parasitoid population dynamics and the importance of these results in the context of biological control and of the study of food webs between aphids and their natural enemies.

Molecular analysis reveals high compartmentalization in aphid-primary parasitoid networks and low parasitoid sharing between crop and noncrop habitats.

The ecosystem service of insect pest regulation by natural enemies, such as primary parasitoids, may be enhanced by the presence of uncultivated, semi-natural habitats within agro-ecosystems, although quantifying such host-parasitoid interactions is difficult. Here, we use rRNA 16S gene sequencing to assess both the level of parasitism by Aphidiinae primary parasitoids and parasitoid identity on a large sample of aphids collected in cultivated and uncultivated agricultural habitats in Western France. We used these data to construct ecological networks to assess the level of compartmentalization between aphid and parasitoid food webs of cultivated and uncultivated habitats. We evaluated the extent to which uncultivated margins provided a resource for parasitoids shared between pest and nonpest aphids. We compared the observed quantitative ecological network described by our molecular approach to an empirical qualitative network based on aphid-parasitoid interactions from traditional rearing data found in the literature. We found that the molecular network was highly compartmentalized and that parasitoid sharing is relatively rare between aphids, especially between crop and noncrop compartments. Moreover, the few cases of putative shared generalist parasitoids were questionable and could be due to the lack of discrimination of cryptic species or from intraspecific host specialization. Our results suggest that apparent competition mediated by Aphidiinae parasitoids is probably rare in agricultural areas and that the contribution of field margins as a source of these biocontrol agents is much more limited than expected. Further large-scale (spatial and temporal) studies on other crops and noncrop habitats are needed to confirm this.

A universal method for the detection and identification of Aphidiinae parasitoids within their aphid hosts.

Molecular methods are increasingly used to detect and identify parasites in their hosts. However, existing methods are generally not appropriate for studying complex host-parasite interactions because they require prior knowledge of species composition. DNA barcoding is a molecular method that allows identifying species using DNA sequences as an identification key. We used DNA amplification with primers common to aphid parasitoids and sequencing of the amplified fragment to detect and identify parasitoids in their hosts, without prior knowledge on the species potentially present. To implement this approach, we developed a method based on 16S rRNA mitochondrial gene and LWRh nuclear gene. First, we designed two primer pairs specific to Aphidiinae (Hymenoptera), the main group of aphid parasitoids. Second, we tested whether the amplified regions could correctly identify Aphidiinae species and found that 61 species were accurately identified of 75 tested. We then determined the ability of each primer pair to detect immature parasitoids inside their aphid host. Detection was earlier for 16S than for LWRh, with parasitoids detected, respectively, 24 and 48 h after egg injection. Finally, we applied this method to assess parasitism rate in field populations of several aphid species. The interest of this tool for analysing aphid-parasitoid food webs is discussed.

Identification of molecular markers for DNA barcoding in the Aphidiinae (Hym. Braconidae).

Reliable identification of Aphidiinae species (Braconidae) is a prerequisite for conducting studies on aphid-parasitoid interactions at the community level. However, morphological identification of Aphidiinae species remains problematic even for specialists and is almost impossible with larval stages. Here, we compared the efficiency of two molecular markers [mitochondrial cytochrome c oxydase I (COI) and nuclear long wavelength rhodopsin (LWRh)] that could be used to accurately identify about 50 species of Aphidiinae that commonly occur in aphid-parasitoid networks in northwestern Europe. We first identified species on a morphological basis and then assessed the consistency of genetic and morphological data. Probably because of mitochondrial introgression, Aphidius ervi and A. microlophii were indistinguishable on the basis of their COI sequences, whereas LWRh sequences discriminated these species. Conversely, because of its lower variability, LWRh failed to discriminate two pairs of species (Aphidius aquilus, Aphidius salicis, Lysiphlebus confusus and Lysiphlebus fabarum). Our study showed that no unique locus but a combination of two genes should be used to accurately identify members of Aphidiinae.

Abdominal sensory equipment involved in external host discrimination in a solitary parasitoid wasp.

Already parasitized hosts are often of poorer quality than healthy hosts. It is therefore usually advantageous for parasitoid females to recognize and reject them. Parasitized hosts can be identified on the basis of various physical or chemical marks present on the surface or inside the hosts or their surroundings in the case of concealed host. Here we studied host discrimination behaviors of females of a certain population of Pachycrepoideus vindemmiae, a solitary ectoparasitoid, which are known to reject large-sized parasitized hosts after an abdominal examination of their surface. We first investigated females' recognition behaviors of host parasitism status when confronted to small-sized hosts (Drosophila melanogaster pupae) as host size may influence the use of different cues for host selection. We showed that, in such a situation, females also discriminate parasitized hosts after an external host exploration with the tip of their ovipositor sheath (third valvulae). We then described the sense organs present on the different parts of the ovipositor by means of scanning and transmission electron microscopy analysis. As the extremity of the third valvulae bears only one type of sensilla that appears to be chemoreceptors, we considered these sensilla as highly likely to be involved in host discrimination in P. vindemmiae. To our knowledge, this is the first time that receptors located on the ovipositor sheath are described as implicated in host discrimination in parasitoid wasps. We discuss potential chemical markers that might be detected by these receptors.

The conflicting relationships between aphids and men: a review of aphid damage and control strategies.

In this review, after giving some figures on the economic impact of aphids on agricultural production, we describe the different mechanisms leading to yield losses (direct damage due to sieve drain and plant reaction, indirect damage, often the most important, due to virus transmission). Then, after a history of chemical control and of its limits, the main control strategies (chemical control with decision rules, plant resistance, biological control, farming practices) are reviewed in the light of an integrated pest management approach. Several topics tackled in this article are exemplified for cereal aphids, which are among the most important in Europe as direct feeders and virus vectors.

Evolutionary ecology of the interactions between aphids and their parasitoids.

Many organisms, including entomopathogenous fungi, predators or parasites, use aphids as ressources. Parasites of aphids are mostly endoparasitoid insects, i.e. insects which lay eggs inside the body of an other insect which will die as a result of their development. In this article, we review the consequences of the numerous pecularities of aphid biology and ecology for their endoparasitoids, notably the Aphidiinae (Hymenoptera: Braconidae). We first examine the various mechanisms used by aphids for defence against these enemies. We then explore the strategies used by aphidiine parasitoids to exploit their aphid hosts. Finally, we consider the responses of both aphids and parasitoids to ecological constraints induced by seasonal cycles and to environmental variations linked to host plants and climate. The fundamental and applied interest of studying these organisms is discussed.

Epicuticular factors involved in host recognition for the aphid parasitoid Aphidius rhopalosiphi.

In insect parasitoids, fitness is dependent on the host finding and recognition abilities of the female. Host recognition cues have been described for various host-parasitoid systems, but are still under investigation in aphid parasitoids. Our study aimed to clarify the respective role of physical and chemical cues in recognition of the aphid cuticle. Shed aphid exuviae were used as an elicitor in order to avoid any influence of color, movement, or volatiles present in a living aphid. We assessed the effect of chemical and heat treatments on the texture of the cuticle by using scanning electron microscopy and tested the recognition of treated cuticles by the parasitoid. We showed that recognition cues of the cuticle can be removed chemically (using combined treatments with n-hexane and methanol). Moreover, heat treatment destroyed the physical texture of the cuticle without significantly reducing parasitoid recognition. In a second step, we showed that epicuticular extracts deposited on pieces of paper triggered female attack behavior. First results concerning the chemical composition of the active extract are presented. This study shows that chemical compounds extractable by organic solvents mediate cuticle recognition by aphid parasitoids.