Four New Species of Pseudephedrus (Hymenoptera, Braconidae, Aphidiinae) from Chile with a Key to Species Identification.

The following work consists of the description of four new species of the genus Pseudephedrus Starý (Aphidiinae), endemic to South America, associated with endemic callaphidid aphid hosts. The descriptions of the new species are based on new samples from Chile and Argentina. The new species described here are as follows: Pseudephedrus staryi sp.n., which is morphologically very close to the already described P. chilensis Starý, with clear apomorphies; P. pubescens sp.n., which presents a setose scutellum, P. flava sp.n. and P. patagonicus sp.n. which have distinct, very elongated, and crenulated petioles representing synapomorphies and probably form separate phylogenetic lineages within Pseudephedrus. All hosts are aphids from the genus Neuquenaphis Blanchard, with varying degrees of diet specialization. From our field and laboratory observations, we hypothesize that, since attempts to sample parasitoids using sweep nets were much more successful than sampling from collected aphids, and since we found mummified aphids only on the ground among the fallen leaves under the trees, Neuquenaphis aphids fall to the ground showing a dropping behaviour as a defence against natural enemies after being stung by parasitoids. This makes rearing from live aphids very unsuccessful and could help explain why it has been difficult to collect and describe species. We supplement the distribution of Pseudephedrus from South America and present a key for the identification of all species based on their morphology.

Composition and Food Web Structure of Aphid-Parasitoid Populations on Plum Orchards in Chile.

By increasing plant diversity in agroecosystems, it has been proposed that one can enhance and stabilize ecosystem functioning by increasing natural enemies' diversity. Food web structure determines ecosystem functioning as species at different trophic levels are linked in interacting networks. We compared the food web structure and composition of the aphid- parasitoid and aphid-hyperparasitoid networks in two differentially managed plum orchards: plums with inter-rows of oats as a cover crop (OCC) and plums with inter-rows of spontaneous vegetation (SV). We hypothesized that food web composition and structure vary between OCC and SV, with network specialization being higher in OCC and a more complex food web composition in SV treatment. We found a more complex food web composition with a higher species richness in SV compared to OCC. Quantitative food web metrics differed significantly among treatments showing a higher generality, vulnerability, interaction evenness, and linkage density in SV, while OCC presented a higher degree of specialization. Our results suggest that plant diversification can greatly influence the food web structure and composition, with bottom-up effects induced by plant and aphid hosts that might benefit parasitoids and provide a better understanding of the activity, abundance, and interactions between aphids, parasitoids, and hyperparasitoids in plum orchards.

Effect of the Genotypic Variation of an Aphid Host on the Endosymbiont Associations in Natural Host Populations.

Understanding the role of facultative endosymbionts on the host's ecology has been the main aim of the research in symbiont-host systems. However, current research on host-endosymbiont dynamics has failed to examine the genetic background of the hosts and its effect on host-endosymbiont associations in real populations. We have addressed the seasonal dynamic of facultative endosymbiont infections among different host clones of the grain aphid Sitobion avenae, on two cereal crops (wheat and oat) and whether their presence affects the total hymenopteran parasitism of aphid hosts at the field level. We present evidence of rapid seasonal shifts in the endosymbiont frequency, suggesting a positive selection of endosymbionts at the host-level (aphids) through an agricultural growing season, by two mechanisms; (1) an increase of aphid infections with endosymbionts over time, and (2) the seasonal replacement of host clones within natural populations by increasing the prevalence of aphid clones closely associated to endosymbionts. Our results highlight how genotypic variation of hosts can affect the endosymbiont prevalence in the field, being an important factor for understanding the magnitude and direction of the adaptive and/or maladaptive responses of hosts to the environment.

Interspecific competition among aphid parasitoids: molecular approaches reveal preferential exploitation of parasitized hosts.

When a guild of species exploit the same limited resources, interspecific competition induces the exclusion of inferior competitors, in which case, interspecific recognition mechanisms are needed. Here, we address resource partitioning and interspecific competition among three main solitary parasitoid species attacking the same host resource, the aphid Sitobion avenae in wheat fields. Optimal host acceptance models predict that parasitoid species should prefer attacking unparasitized hosts when they are available in order to maximize their fitness, as already parasitized hosts are less valuable for laying eggs, especially for inferior competitors. Therefore, we expected the level of competition (multiparasitism) in the field to increase at low host density. By using a combination of taxonomical (determination) and molecular (PCR-based) approaches, we assessed the species of all parasitoid adults and immature stages within aphid hosts. Our results demonstrate that, early in the season, the multiparasitism rates were low, whereas they were high in the mid-late season, corresponding to an aphid density decrease over time. Moreover, parasitoid species could not have been exploiting host resources randomly and the better competitor, Aphidius ervi, seemed to be foraging preferentially on hosts already parasitized by the inferior competitor A. rhopalosiphi, even when unparasitized hosts were still available. This could be due to differences in their host detection capability, as species with a narrow host range may be better at detecting their hosts in comparison with species with a greater host range, such as A. ervi, with a greater host range within the guild. Our study suggests differences in the host exploitation of two prevalent parasitoid species through the main period of aphid colonization, which still allowed the coexistence of a third inferior competitor (A. rhopalosiphi) within the assemblage, in spite of some negative interactions (multiparasitism) and redundancies.

Low bacterial community diversity in two introduced aphid pests revealed with 16S rRNA amplicon sequencing.

Bacterial endosymbionts that produce important phenotypic effects on their hosts are common among plant sap-sucking insects. Aphids have become a model system of insect-symbiont interactions. However, endosymbiont research has focused on a few aphid species, making it necessary to make greater efforts to other aphid species through different regions, in order to have a better understanding of the role of endosymbionts in aphids as a group. Aphid endosymbionts have frequently been studied by PCR-based techniques, using species-specific primers, nevertheless this approach may omit other non-target bacteria cohabiting a particular host species. Advances in high-throughput sequencing technologies are complementing our knowledge of microbial communities by allowing us the study of whole microbiome of different organisms. We used a 16S rRNA amplicon sequencing approach to study the microbiome of aphids in order to describe the bacterial community diversity in introduced populations of the cereal aphids, Sitobion avenae and Rhopalosiphum padi in Chile (South America). An absence of secondary endosymbionts and two common secondary endosymbionts of aphids were found in the aphids R. padi and S. avenae, respectively. Of those endosymbionts, Regiella insecticola was the dominant secondary endosymbiont among the aphid samples. In addition, the presence of a previously unidentified bacterial species closely related to a phytopathogenic Pseudomonad species was detected. We discuss these results in relation to the bacterial endosymbiont diversity found in other regions of the native and introduced range of S. avenae and R. padi. A similar endosymbiont diversity has been reported for both aphid species in their native range. However, variation in the secondary endosymbiont infection could be observed among the introduced and native populations of the aphid S. avenae, indicating that aphid-endosymbiont associations can vary across the geographic range of an aphid species. In addition, we discuss the potential role of aphids as vectors and/or alternative hosts of phytopathogenic bacteria.

Diversity, frequency, and geographic distribution of facultative bacterial endosymbionts in introduced aphid pests.

Facultative bacterial endosymbionts in insects have been under intense study during the last years. Endosymbionts can modify the insect's phenotype, conferring adaptive advantages under environmental stress. This seems particularly relevant for a group of worldwide agricultural aphid pests, because endosymbionts modify key fitness-related traits, including host plant use, protection against natural enemies and heat tolerance. Aimed to understand the role of facultative endosymbionts on the success of introduced aphid pests, the distribution and abundance of 5 facultative endosymbionts (Hamiltonella defensa, Regiella insecticola, Serratia symbiotica, Rickettsia and Spiroplasma) were studied and compared in 4 cereal aphids (Sitobion avenae, Diuraphis noxia, Metopolophium dirhodum and Schizaphis graminium) and in the pea aphid Acyrthosiphon pisum complex from 2 agroclimatic zones in Chile. Overall, infections with facultative endosymbionts exhibited a highly variable and characteristic pattern depending on the aphid species/host race and geographic zone, which could explain the success of aphid pest populations after their introduction. While S. symbiotica and H. defensa were the most frequent endosymbionts carried by the A. pisum pea-race and A. pisum alfalfa-race aphids, respectively, the most frequent facultative endosymbiont carried by all cereal aphids was R. insecticola. Interestingly, a highly variable composition of endosymbionts carried by S. avenae was also observed between agroclimatic zones, suggesting that endosymbionts are responding differentially to abiotic variables (temperature and precipitations). In addition, our findings constitute the first report of bacterial endosymbionts in cereal aphid species not screened before, and also the first report of aphid endosymbionts in Chile.

Does sex-biased dispersal account for the lack of geographic and host-associated differentiation in introduced populations of an aphid parasitoid?

Host recognition and use in female parasitoids strongly relies on host fidelity, a plastic behavior which can significantly restrict the host preferences of parasitoids, thus reducing the gene flow between parasitoid populations attacking different insect hosts. However, the effect of migrant males on the genetic differentiation of populations has been frequently ignored in parasitoids, despite its known impact on gene flow between populations. Hence, we studied the extent of gene flow mediated by female and male parasitoids by assessing sibship relationships among parasitoids within and between populations, and its impact on geographic and host-associated differentiation in the aphid parasitoid Aphidius ervi. We report evidences of a high gene flow among parasitoid populations on different aphid hosts and geographic locations. The high gene flow among parasitoid populations was found to be largely male mediated, suggested by significant differences in the distribution of full-sib and paternal half-sib dyads of parasitoid populations.

Adaptive evolution of a generalist parasitoid: implications for the effectiveness of biological control agents.

The use of alternative hosts imposes divergent selection pressures on parasitoid populations. In response to selective pressures, these populations may follow different evolutionary trajectories. Divergent natural selection could promote local host adaptation in populations, translating into direct benefits for biological control, thereby increasing their effectiveness on the target host. Alternatively, adaptive phenotypic plasticity could be favored over local adaptation in temporal and spatially heterogeneous environments. We investigated the existence of local host adaptation in Aphidius ervi, an important biological control agent, by examining different traits related to infectivity (preference) and virulence (a proxy of parasitoid fitness) on different aphid-host species. The results showed significant differences in parasitoid infectivity on their natal host compared with the non-natal hosts. However, parasitoids showed a similar high fitness on both natal and non-natal hosts, thus supporting a lack of host adaptation in these introduced parasitoid populations. Our results highlight the role of phenotypic plasticity in fitness-related traits of parasitoids, enabling them to maximize fitness on alternative hosts. This could be used to increase the effectiveness of biological control. In addition, A. ervi females showed significant differences in infectivity and virulence across the tested host range, thus suggesting a possible host phylogeny effect for those traits.