Assessment of Sublethal and Transgenerational Effects of Pirimicarb on the Wheat Aphids Rhopalosiphum padi and Sitobion avenae

The wheat aphids, Rhopalosiphum padi (Linnaeus) and Sitobion avenae (Fabricius), are key pests on wheat crops worldwide. Management practices rely primarily on insecticides. The pirimicarb (carbamate) is used extensively as an effective insecticide to control these two aphids. In addition to the mortality caused by pirimicarb, various sublethal effects may occur in aphids when exposed to low lethal or sublethal doses. Understanding the general effect of pirimicarb on aphids could help increasing rational use of this insecticide. Under laboratory conditions, we assessed the sublethal effects of a low lethal concentration of pirimicarb (LC₂₅) on biological traits and acetylcholinesterase (AChE) activity of R. padi and S. avenae. Both direct and transgenerational effects, i.e. on parent and the F1 generations were assessed, respectively. We found that R. padi and S. avenae responded differentially to the LC₂₅ of pirimicarb. The parent generation of R. padi showed a 39% decrease in fecundity and multiple transgenerational effects were observed in the F1 generation; overall juvenile development, reproductive period, adult longevity and lifespan were longer than those of the control group. By contrast, LC₂₅ of pirimicarb showed almost no effects on S. avenae biological traits in both the parent and F1 generations; only the pre-reproductive duration was reduced in F1 generations. Demographic parameter estimates (e.g. r_m) showed similar trend, i.e. significant negative effect on R. padi population growth and no effect on S. avenae. However, AChE activity decreased in both R. padi and S. avenae treated by the LC₂₅ of pirimicarb. We demonstrated sublethal and transgenerational effects of pirimicarb in the two wheat aphid species; it hinted at the importance of considering sublethal effects (including hormesis) of pirimicarb for optimizing Integrated Pest Management (IPM) of wheat aphids.     

Sublethal effects of selected insecticides on fecundity and wing dimorphism of green peach aphid (Hom., Aphididae)

Effects of sublethal concentrations (LC₂₅) of six insecticides (imidacloprid, rotenone, fenvalerate, abamectin, pirimicarb and azadirachtin) on fecundity and wing dimorphism of the green peach aphid, Myzus persicae (Sulzer), were studied both under laboratory and greenhouse conditions. In the laboratory, aphid reproduction reduced by 44.29% and 54.01% when rotenone and abamectin treatments were applied at sublethal dose, respectively, and sublethal fenvalerate application resulted in markedly lower average reproduction per female per day compared with control. Reproductive duration of aphid treated with abamectin significantly decreased by 44.19%. But in the greenhouse, there were no evident differences in the aphid fecundity and reproductive duration between treatments and control. Life‐table parameters also demonstrated that the six insecticides at sublethal doses did not stimulate the aphid reproductive potential. In the laboratory, after being exposed to sublethal doses of imidacloprid and fenvalerate, the proportions of alate progeny in aphid progeny were significantly higher than that of the control. In the greenhouse, percentages of alate offspring from the mother aphids treated with imidacloprid, fenvalerate and abamectin increased pronouncedly compared with control. Mortality rates of offspring in the nymphal stages from adults treated with insecticides revealed no significant changes between laboratory and greenhouse. The developmental time in days of the offspring varied in all treatments. Mechanisms of insecticide‐induced resurgence are discussed.     

Feeding response of the cotton aphid,Aphis gossypii,to sublethal rates of flonicamid and imidacloprid

The cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is an important sap‐sucking pest of many plants, including melons and peppers. This study was conducted to determine the effects of sublethal exposure to flonicamid and imidacloprid and the mechanisms by which these insecticides affect the feeding behavior of A. gossypii. The median lethal concentrations (LC₅₀) of flonicamid and imidacloprid for adult A. gossypii were 2.40 and 1.92 mg l^?1, respectively. The lower lethal concentrations of flonicamid were 1.01 mg l^?1 (LC₃₀) and 0.29 mg l^?1 (LC₁₀), and those of imidacloprid were 0.82 mg l^?1 (LC₃₀) and 0.24 mg l^?1 (LC₁₀). The developmental period of A. gossypii nymphs at LC₃₀ was 3.6 days for both insecticides, which was shorter than that of the untreated controls (4.2 days). Longevity and total fecundity of A. gossypii adults were decreased at the sublethal concentrations of both insecticides. The lowest net reproductive rate was observed in A. gossypii treated with the LC₃₀ of flonicamid. Feeding behavior analyses using an electrical penetration graph showed that sublethal concentrations of flonicamid and imidacloprid had significant effects on the duration of phloem ingestion. Higher doses of flonicamid induced starvation by inhibiting phloem ingestion, whereas imidacloprid acted as a contact toxin rather than an inhibitor of feeding behavior.     

The effect of insecticides on the distribution of foracing parasitoids,Diaeretiella rapae [Hym.: Braconidae] on plants

Foraging aphid parasitoids,Diaeretiella rapae M'Intosh, were exposed to sublethal doses of the insecticides pirimicarb, permethrin and malathion on brusslls sprouts plants. Observations on wasp distribution over time revealed that wasps spent less time on sprayed plants, relative to controls and, while on these plants, tended to concentrate activity on unsprayed surfaces. For permethrin and malathion, pesticide residues reversed the stereotypic upward foraging pattern of the wasp. Negative consequences of sublethal pesticide doses for parasitoid foraging efficiency are discussed.      

A comparison of life history traits of sexual and asexual strains of the parasitoid wasp,Lysiphlebus fabarum (Braconidae: Aphidiinae)

1. This study examined biological characteristics of sexual and asexual strains of the parasitoid wasp, Lysiphlebus fabarum (Marshall) (Hymenoptera: Braconidae). 2. Strains were reared in different instar hosts (the black bean aphid, Aphis fabae Scopoli) under identical environmental conditions (21 °C, 65–75% RH, and LD 16:8 h). 3. Results showed that the second instar of the aphid is the most suitable growth stage for both strains, as the wasps that emerged from the second instar hosts were larger, more fecund, and had larger egg size. Trade‐offs between the fitness components of the parasitoid were clearer when the parasitoids were reared in suboptimal instars. 4. According to the results, sexual females emerged around 1 day earlier and lived around 0.5 day less than asexual females. Also, sexual females emerged with a lower initial egg load, although these wasps tend to have larger eggs than asexual females. Asexual females may enjoy greater longevity and higher developmental plasticity which suggests a higher degree of synchronization with pest population dynamism. 5. The results suggest that sexual wasps, in contrast to asexual wasps, invest more in egg size than in egg load. This study suggests strain‐specific adaptations of L. fabarum to different instars of the black bean aphid by which the allocation of nutritional resources to various functions differs between strains. 6. Furthermore, differences in life history traits between strains can greatly influence the population dynamics of each strain, and hence their effectiveness in suppressing pest populations.     

Sublethal effects of imidacloprid exposure on <Emphasis Type="Italic">Spalangia endius</Emphasis>, a pupal parasitoid of filth flies

Parasitoids and neonicotinoids can both suppress economically harmful filth fly populations. However, sublethal effects of neonicotinoids have not previously been studied for commonly used species of filth fly parasitoids. Exposure to an LC₅₀ of imidacloprid decreased the ability of surviving individuals of the parasitoid wasp Spalangia endius Walker (Hymenoptera: Pteromalidae) to kill house fly pupae under some conditions. In an unburied-hosts experiment, significantly more flies and fewer parasitoids emerged in the LC₅₀ imidacloprid treatment versus the LC₁₀ or controls. Parasitoid sex ratio and longevity were not affected. However, in a buried-hosts experiment, parasitoid and fly emergence were independent of treatment. ELISA (enzyme-linked immunosorbent assay) showed lower imidacloprid residues in or on parasitoids exposed to the media in which hosts were buried. Our findings suggest that substrate may reduce pesticides on biological control agents that burrow, making them more effective.     

Body size affects host defensive behavior and progeny fitness in a parasitoid wasp,Lysiphlebus fabarum

This study tested effects of maternal body size on foraging behavior and progeny development in a thelytokous population of Lysiphlebus fabarum (Marshall) (Hymenoptera: Braconidae). Small and large wasps were reared from first and second instar hosts [black bean aphid, Aphis fabae Scopoli (Hemiptera: Aphididae)], respectively, and each was provided with a patch (bean leaf disk) containing either 15 small (second instar) or 15 large (fourth instar) hosts for a 30‐min foraging period. Neither body size nor host size affected time allocation to various behaviors within a patch, but second instar aphids produced significantly more mummies than fourth instars. The preferred attack orientation was from the side of the aphid, suggesting wasps were sensitive to the risk of smearing with cornicle secretions. Few wasps developed in fourth instar hosts, suggesting later host instars were somewhat resistant to parasitism. Second instar hosts, the most suitable stage for L. fabarum development, relied more on defensive behavior, specifically kicking and secreting cornicle droplets. Large wasps were more likely to elicit a double cornicle secretion, indicating that aphids graded their response to the size of the attacker. Larger wasps were also more likely to be smeared with cornicle secretion, suggesting they were more vulnerable than small wasps. Although small wasps had smaller eggs than large wasps, there was no effect of maternal egg size on the size of progeny. However, daughters of small females emerged with larger egg loads than daughters of large mothers, and their eggs tended to be slightly smaller, although not significantly. Regression analysis revealed a positive correlation between maternal egg size and progeny developmental time for small and large wasps, and between maternal egg size and progeny egg load for small wasps. These results confirm maternal effects of body size in an aphid parasitoid, and reveal that vulnerability to host behavioral defenses is also body size dependent.     

Influence of aphid size,age and behaviour on host choice by the parasitoid wasp Ephedrus californicus: a test of host-size models

Summary When host quality varies, parasitoid wasps are expected to oviposit selectively in high-quality hosts. We tested the assumption underlying host-size models that, for solitary species of wasps, quality is based on host size. Using Ephedrus californicus, a solitary endoparasitoid of the pea aphid, we evaluated the influence of aphid size (= mass), age and defensive behaviours on host selection. Experienced parasitoid females were given a choice among three classes of 5-day-old apterous nymphs: small aphids that had been starved daily for 4 h (S4) and 6 h (S6) respectively, and large aphids permitted to feed (F) normally. Wasps attacked more, and laid more eggs in, small than large aphids (S6>S4>F). This rank-order for attack did not change when females could choose among aphids of the same size that differed in age; however, wasps oviposited in all attacked aphids with equal probability. Host size did not influence parasitoid attack rates when aphids were anaesthetized so that they could not escape or defend themselves. As predicted by host-size models, wasp size increased with host size (F>S4; S6), but large wasps required longer to complete development than their smaller counterparts (S4E. californicus reflects a trade-off between maximization of fitness gains per egg and the economics of search-time allocation. Because large aphids are more likely to escape parasitization, a wasp must balance her potential gain in fitness by ovipositinng in a high-quality (large) aphid against her potential cost in terms of lost opportunity time if the attack fails.     

Quantitative trait locus analysis of parasitoid counteradaptation to symbiont-conferred resistance

Insect hosts and parasitoids are engaged in an intense struggle of antagonistic coevolution. Infection with heritable bacterial endosymbionts can substantially increase the resistance of aphids to parasitoid wasps, which exerts selection on parasitoids to overcome this symbiont-conferred protection (counteradaptation). Experimental evolution in the laboratory has produced counteradapted populations of the parasitoid wasp Lysiphlebus fabarum. These populations can parasitize black bean aphids (Aphis fabae) protected by the bacterial endosymbiont Hamiltonella defensa, which confers high resistance against L. fabarum. We used two experimentally evolved parasitoid populations to study the genetic architecture of the counteradaptation to symbiont-conferred resistance by QTL analysis. With simple crossing experiments, we showed that the counteradaptation is a recessive trait depending on the maternal genotype. Based on these results, we designed a customized crossing scheme to genotype a mapping population phenotyped for the ability to parasitize Hamiltonella-protected aphids. Using 1835 SNP markers obtained by ddRAD sequencing, we constructed a high-density linkage map consisting of six linkage groups (LGs) with an overall length of 828.3 cM and an average marker spacing of 0.45 cM. We identified a single QTL associated with the counteradaptation to Hamiltonella in L. fabarum on linkage group 2. Out of 120 genes located in this QTL, several genes encoding putative venoms may represent candidates for counteradaptation, as parasitoid wasps inject venoms into their hosts during oviposition.Subject terms: Experimental evolution, Evolutionary genetics, Evolutionary ecology, Genetic linkage study     

三种杀虫剂对麦田蚜虫和天敌的影响

通过对施用杀虫剂吡虫啉、抗蚜威、广谱性杀虫剂氧化乐果对麦田蚜虫和天敌的影响进行分析 ,结果表明 ,施用杀虫剂对麦田蚜虫防效高 ,对其天敌有保护作用 ,且瓢蚜比降低。使用 1 0 %吡虫啉( 1 0g 667m2 )后 5～ 2 5天瓢蚜比为 1∶34～ 1∶1 70 ;用 50 %抗蚜威 ( 5g 667m2 )后 1 0～ 2 0天瓢蚜比为 1∶31～1∶1 95;而广谱性杀虫剂氧化乐果 ( 50mL 667m2 )对麦田蚜虫防效好 ,对天敌杀伤力大 ,药后 1 5天瓢蚜比为1∶2 65。施用化学农药可使蚜茧蜂寄生率提高。     

Development of neonicotinoid resistance in the cotton aphid Aphis gossypii (Hemiptera: Aphididae) in Japan

A neonicotinoid-resistant Aphis gossypii Glover population was first detected from cucumber and sweet pepper crops in Miyazaki Prefecture, Japan, in April 2012. In this study, we determined the insecticide susceptibility of five field-collected populations of A. gossypii to seven neonicotinoids. Insecticide susceptibility was determined by performing bioassays and using the seedling-treatment method. Insecticide susceptibility of a susceptible population of A. gossypii, originally collected from a cucumber field in 2008, was also determined. High mortality (96.4–100 %) was observed in the susceptible population for the seven insecticides. In contrast, the mortality rate of the five field-collected populations collected in 2012 was low for five of the seven neonicotinoid insecticides tested: imidacloprid (26.7–65.5 %), dinotefuran (0–27.3 %), clothianidin (20.0–35.7 %), thiamethoxam (7.1–42.3 %), and nitenpyram (6.7–32.1 %). Mortality was 86.2–100 % for acetamiprid and 90.2–100 % for thiacloprid. The resistance ratio in comparison with the susceptible population was the highest for clothianidin (687-fold), lowest for thiacloprid (17-fold), and ranged from 43- to 253-fold for the other five neonicotinoids. Thus, the frequency of insecticide use needs to be reduced and other insecticides need to be employed in order to prevent the development of resistance to these two insecticides in the future.     

Sub-lethal Effects of the Insecticides Pirimicarb and Dimethoate on the Aphid Parasitoid Diaeretiella rapae (Hymenoptera: Braconidae) When Attacking and Developing in Insecticide-Resistant Hosts

The effectiveness of parasitoids as biological control agents can be constrained by insecticide use, not only through direct mortality but also as a result of sublethal effects. Several pest aphids have become resistant to a range of insecticides and a resistant strain of Myzus persicae was used in laboratory experiments to investigate sublethal effects of the insecticides pirimicarb and dimethoate on the parasitoid Diaeretiella rapae . Both insecticides produced sublethal effects on D. rapae when the parasitoid attacked and developed in aphids that had been dipped in insecticide solutions. Dimethoate affected oviposition behaviour; females were apparently repelled by residues on the surface of dipped aphids, thus reducing their attack rate and hence the number of mummies produced. Also, the reproductive performance of parasitoids that had developed in pirimicarb-dipped aphids appeared to be adversely affected, in comparison with parasitoids that emerged from uncontaminated hosts, and this was reflected both in lower mummy production and lower attack rates. Pirimicarb, but not dimethoate, affected the sex ratio of the offspring of D. rapae that had developed in dipped aphids, causing a significant increase in the proportion of males. This only occurred when the male parent had developed in a pirimicarb-dipped aphid, suggesting that the effect involves male sterility or mating behaviour, although they appeared to mate normally. These sublethal effects are potential constraints on the efficiency and effectiveness of D. rapae as a biological control agent of aphid pests, but to assess fully their potential impact further studies need to be done using more realistic extended laboratory and semi-field techniques.     

Potential risks of systemic imidacloprid to parasitoid natural enemies of a cerambycid attacking Eucalyptus

A community of insect herbivores has established on eucalyptus species in California following their introduction from Australia. A number of the species are under complete or partial biological control. A response to introduction of additional pest species into the complex has been the application of systemic insecticides to infested trees. Natural enemies that have been introduced to control the various pest species feed on the nectar of treated trees, and thus may be affected by these pesticides. In this study, Eucalyptus rudis trees were treated at label rates with the neonicotinoid systemic insecticide imidacloprid and nectar sampled at 5 months post-treatment during the spring bloom. The concentration of imidacloprid and its toxic metabolites in nectar was measured by ELISA at 660 ppb. Adults of the encyrtid egg parasitoid Avetianella longoi that were fed floral nectar collected from treated trees had significantly lower survival and reproductive fitness than adults fed nectar from untreated trees. In feeding bioassays, in which the adults were fed a range of concentrations of imidacloprid in sugar water, the LC₅₀ for A. longoi was 212 ppb imidacloprid. Bioassays were also conducted with the braconid larval parasitoid, Syngaster lepidus. The LC₅₀ for S. lepidus was 288 ppb imidacloprid. The insecticide appears to be concentrating in the nectar at higher levels than reported from other plant species and at concentrations exceeding the LC₅₀ for two important parasitoids. If tree treatments become widespread as a result of continual introductions of new eucalypt herbivores, established biological control programs could be at significant risk.     

The impact of insecticide resistance in the currant-lettuce aphid, Nasonovia ribisnigri, on pest management in lettuce

Abstract 1 This paper reports on experiments to determine how two different insecticide resistance phenotypes in the aphid Nasonovia ribisnigri (Mosley), which is a major pest of lettuce, change its susceptibility to pyrethroid insecticides and the carbamate pirimicarb. 2 A novel statistical approach determined how the effectiveness of different insecticides was changed by the two resistance phenotypes. This compared the between‐plant distribution of aphid numbers, as opposed to the mean number of aphids per plant. 3 Results from field cage experiments showed that the effect of the resistances differed. Pyrethroid resistance resulted in lower mortality immediately after application of pyrethroids, whereas resistance to pirimicarb shortened the time over which the chemical was effective. 4 The results of laboratory bioassays suggested that these two resistances were not found together in N. ribisnigri. However, the results reported here contradict this assertion. 5 Experiments with insecticide residues showed that reproduction of resistant N. ribisnigri was greater than that of susceptible N. ribisnigri on plants with ageing insecticide residues, even in circumstances where mortality of resistant and susceptible clones of N. ribisnigri were similar. 6 If more than a few aphids are found on a plant then a whole consignment can be rejected for processing. The results reported here suggest that the effect of both insecticide resistances in N. ribsinigri will be to increase the proportion of lettuce heads with an unacceptable number of aphids on them, leading to increased rejection of plants for processing.     

Host instar suitability of Aphis gossypii (Hemiptera: Aphididae) for Lysiphlebus testaceipes (Hymenoptera: Braconidae) and parasitism effect on aphid life table

Lysiphlebus testaceipes (Cresson) (Hymenoptera: Braconidae, Aphidiinae) has constituted a well-studied parasitoid insect model, but very little is known about the host-instar suitability of aphid for the wasp so far. One of the hosts of L. testaceipes is Aphis gossypii Glover (Hemiptera: Aphididae). The latter is a serious aphid pest to vegetable production in Benin. Therefore, the objectives of our study were to: (1) examine the oviposition behavior of L. testaceipes on A. gossypii; (2) investigate the host-instar suitability of A. gossypii for L. testaceipes; and (3) compare the life table parameters of A. gossypii with aphids parasitized by L. testaceipes and unparasitized aphids (control). The study was conducted in a laboratory at 26 ± 1 °C in petri dishes and revealed that the parasitoid utilized up to seven stabbing stings to handle and oviposit, particularly in older A. gossypii. In aphids parasitized at the third instar, the net reproductive rate R _o as well as the intrinsic rate of natural increase r _m was significantly lower (2.119 ± 0.272 and 0.110 ± 0.018) compared to the control (15.529 ± 1.287 and 0.272 ± 0.008), respectively (p < 0.01).     

Susceptibility of the woolly apple aphid parasitoid,Aphelinus mali (Hym.: Aphelinidae), to common pesticides used in apple orchards in Israel

The effects of one acaricide (cyhexatin), two fungicides (penconazole and sulfur), and six insecticides (azinphos-methyl, chlorpyrifos, imidacloprid, pirimicarb, triazamate and vamidothion) on the adult stage of the parasitoidAphelinus mali (Haldeman), a parasite of the woolly apple aphidEriosoma lanigerum (Hausmann), were investigated under laboratory conditions. Chlorpyrifos (an organophosphorus insecticide-OP) was found to be highly toxic to the adult wasps. Vamidothion (OP) was more toxic to the parasitoid than azinphos-methyl (OP). On the other hand, both chlorpyrifos and azinphos-methyl were found to be harmless to the immature stages of the parasitoid in a test conducted under semi-field conditions. Of the other insecticides, imidacloprid was more toxic to the adult parasitoid than pirimicarb and triazamate. Neither cyhexatin nor penconazole had a considerable toxic effect upon the parasitoid. In contrast, sulfur was found to be moderately toxic to the parasitoid under laboratory conditions, as well as in a field survey in which we followed populations ofE. lanigerum andA. mali parasitization in sulfur-treated plots, and in a plot treated withAmpelomyces quisqualis Ces., a fungus antagonist to powdery mildew. The implications of these results to IPM programs of apple orchards in Israel are discussed     

Development of methods to evaluate susceptibility of soybean aphid to imidacloprid and thiamethoxam at lethal and sublethal concentrations

The soybean aphid, Aphis glycines Matsumura (Homoptera: Aphididae), is a recent introduction (2000) from Asia and has become a serious soybean [Glycine max (L.) Merr. (Fabaceae)] pest in North America. Seed treatments using the neonicotinoid insecticides, imidacloprid and thiamethoxam, have been suggested as a method of control, and the use of these insecticides is becoming widespread. As a consequence, there is increased potential to select for resistance to these compounds. In the case of soybean aphids, baseline susceptibility to neonicotinoid insecticides and standardized methods for bioassay are lacking. A bioassay technique that uses excised soybean leaves immersed in an insecticide solution was developed to determine systemic insecticidal activity at lethal and sublethal concentrations. Mortality and population growth inhibition were evaluated after 7 days. Life table parameters were calculated by exposing 1‐day‐old aphids to three concentrations of thiamethoxam. Aphid mortality and nymph production were recorded daily until the entire cohort collapsed. Soybean aphid age‐specific survivorship, fecundity, net reproductive rate, longevity, intrinsic rate of increase, discrete daily growth rate, and life expectancy were all significantly reduced at higher thiamethoxam concentrations. Soybean aphid response to both insecticides was similar, and both compounds were very toxic with LC₅₀s of 31.3 and 16.9 ng ml^?1 and EC₅₀s of 6.3 and 5.4 ng ml^?1 for imidacloprid and thiamethoxam, respectively. These results indicate that the methods developed in this study had negligible impact on the life table estimates measured and can be used to develop a baseline of susceptibility as a benchmark for subsequent resistance monitoring. Given the rapid and widespread adoption of this new insecticide class, vigilant monitoring for changes in susceptibility will be essential to its long‐term sustainability.     

Symbiont‐conferred protection against Hymenopteran parasitoids in aphids: how general is it?

1. Hosts are often targeted by multiple species of parasites, leading to a confluence of selective pressures on them. In response, hosts may either evolve defences that act very generally, or specific defences against particular parasites. Aphids are attacked by multiple species of endoparasitoid wasps, and there is clear evidence that heritable endosymbionts can confer resistance against some of these wasps. Less clear is how symbiont‐conferred resistance in a single host acts against multiple parasitoid species. 2. This question was addressed in the black bean aphid, Aphis fabae (Scopoli). Unprotected aphids and aphids protected by three different strains of the defensive endosymbiont Hamiltonella defensa were exposed to four species of parasitic wasps: the parthenogenetic species Lysiphlebus fabarum (Marshall), which was represented by three different asexual lines, and the sexual species Aphidius colemani (Viereck), Binodoxys angelicae (Halliday), and Aphelinus chaonia (Walker). 3. Hamiltonella defensa provided strong protection against L. fabarum and Aphidius colemani, but there was no evidence that H. defensa‐infected aphids were more resistant to the other parasitoid species. While Aphidius colemani was virtually unable to parasitise any aphids harbouring H. defensa, there was variation among the three asexual lines of L. fabarum in how susceptible they were to the defence provided by the different symbiont strains, resulting in a significant genotype‐by‐genotype interaction. 4. The present results suggest that symbiosis with H. defensa does not provide aphids with a general defence against parasitoid wasps, possibly because some species have evolved specific counter adaptations or because biological differences preclude the symbiont's effectiveness against these species.     

Does variation in host plant association and symbiont infection of pea aphid populations induce genetic and behaviour differentiation of its main parasitoid, Aphidius ervi?

The host-associated differentiation (HAD) hypothesis states that higher trophic levels in parasitic associations should exhibit similar divergence in case of host sympatric speciation. We tested HAD on populations of Aphidius ervi the main parasitoid of the pea aphid Acyrthosiphon pisum, emerging from host populations specialized on either alfalfa or red clover. Host and parasitoid populations were assessed for genetic variation and structure, while considering geography, host plant and host aphid protective symbionts Regiella insecticola and Hamiltonella defensa as potential covariables. Cluster and hierarchical analyses were used to assess the contribution of these variables to population structure, based on genotyping pea aphids and associated A. ervi with microsatellites, and host aphid facultative symbionts with 16S rDNA markers. Pea aphid genotypes were clearly distributed in two groups closely corresponding with their plant origins, confirming strong plant associated differentiation of this aphid in North America. Overall parasitism by A. ervi averaged 21.5 % across samples, and many parasitized aphids producing a wasp hosted defensive bacteria, indicating partial or ineffective protective efficacy of these symbionts in the field. The A. ervi population genetic data failed to support differentiation according to the host plant association of their pea aphid host. Potential for parasitoid specialization was also explored in experiments where wasps from alfalfa and clover aphids were reciprocally transplanted on alternate hosts, the hypothesis being that wasp behaviour and parasitic stages should be most adapted to their host of origin. Results revealed higher probability of oviposition on the alfalfa aphids, but higher adult emergence success on red clover aphids, with no interaction as expected under HAD. We conclude that our study provides no support for the HAD in this system. We discuss factors that might impair A. ervi specialization on its divergent aphid hosts on alfalfa and clover.