Ecological Niche Models and Coalescent Analysis of Gene Flow Support Recent Allopatric Isolation of Parasitoid Wasp Populations in the Mediterranean

Background

The integration of multiple complementary approaches is a powerful way to understand the processes of diversification and speciation. The parasitoid wasp Aphidius transcaspicus Telenga (Hymenoptera: Braconidae) is a parasitoid of Hyalopterus aphids across a wide geographic range. This species shows a remarkable degree of genetic structure among western, central, and eastern Mediterranean population clusters. In this paper we attempt to better characterize this genetic structure.

Methodology/Principal Findings

We use a Bayesian coalescent analysis of gene flow under the Isolation with Migration model using mitochondrial and microsatellite markers together with climate-based ecological niche models to better understand the genetic structure of A. transcaspicus in the Mediterranean. The coalescent analysis revealed low levels of migration among western and eastern Mediterranean populations (Nm<1) that were not statistically distinguishable from zero. Niche models showed that localities within population clusters each occupy areas of continuously high environmental suitability, but are separated from each other by large regions of completely unsuitable habitat that could limit dispersal. Overall, environmental characteristics were similar among the population clusters, though significant differences did emerge.

Conclusions/Significance

These results support contemporary allopatric isolation of Mediterranean populations of A. transcaspicus, which together with previous analyses indicating partial behaviorally mediated reproductive isolation, suggest that the early stages of cryptic speciation may be in progress.     

One step ahead: a parasitoid disperses farther and forms a wider geographic population than its fig wasp host

The structure of populations across landscapes influences the dynamics of their interactions with other species. Understanding the geographic structure of populations can thus shed light on the potential for interacting species to co‐evolve. Host–parasitoid interactions are widespread in nature and also represent a significant force in the evolution of plant–insect interactions. However, there have been few comparisons of population structure between an insect host and its parasitoid. We used microsatellite markers to analyse the population genetic structure of Pleistodontes imperialis sp. 1, a fig‐pollinating wasp of Port Jackson fig (Ficus rubiginosa), and its main parasitoid, Sycoscapter sp. A, in eastern Australia. Besides exploring this host–parasitoid system, our study also constitutes, to our knowledge, the first study of population structure in a nonpollinating fig wasp species. We collected matched samples of pollinators and parasitoids at several sites in two regions separated by up to 2000 km. We found that pollinators occupying the two regions represent distinct populations, but, in contrast, parasitoids formed a single population across the wide geographic range sampled. We observed genetic isolation by distance for each species, but found consistently lower F_ST and R_ST values between sites for parasitoids compared with pollinators. Previous studies have indicated that pollinators of monoecious figs can disperse over very long distances, and we provide the first genetic evidence that their parasitoids may disperse as far, if not farther. The contrasting geographic population structures of host and parasitoid highlight the potential for geographic mosaics in this important symbiotic system.     

Molecular Markers for Identification of the Hyperparasitoids Dendrocerus carpenteri and Alloxysta xanthopsis in Lysiphlebus testaceipes Parasitizing Cereal Aphids

Polymerase chain reaction (PCR)-based molecular markers have been developed to detect the presence of primary parasitoids in cereal aphids and used to estimate primary parasitism rates. However, the presence of secondary parasitoids (hyperparasitoids) may lead to underestimates of primary parasitism rates based on PCR markers. This is because even though they kill the primary parasitoid, it’s DNA can still be amplified, leading to an erroneous interpretation of a positive result. Another issue with secondary parasitoids is that adults are extremely difficult to identify using morphological characters. Therefore, we developed species-specific molecular markers to detect hyperparasitoids. A 16S ribosomal RNA mitochondrial gene fragment was amplified by PCR and sequenced from two secondary parasitoid species, Dendrocerus carpenteri (Curtis) (Hymenoptera: Megaspilidae) and Alloxysta xanthopsis (Ashmead) (Hymenoptera: Charipidae), four geographic isolates of the primary parasitoid, Lysiphlebus testaceipes (Cresson) (Hymenoptera: Braconidae), and six aphid species common to cereal crops. Species-specific PCR primers were designed for each insect on the basis of these 16S rRNA gene sequences. Amplification of template DNA, followed by agarose gel electrophoresis, successfully distinguished D. carpenteri and A. xanthopsis from all four isolates of L. testaceipes and all six cereal aphid species in this laboratory test.     

COMPARATIVE GEOGRAPHIC STRUCTURES OF TWO PARASITOID-HOST INTERACTIONS

Recent studies of parasitoid-host interactions have demonstrated that parasitoids and their hosts are geographically structured for traits such as virulence and encapsulation defenses, but no studies have yet compared the geographic structure of parasitoids and hosts using neutral genetic markers. Such studies of geographic structure are needed to evaluate the underlying geographic scale at which these interactions evolve and allow assessment of the relative effects of selection and gene flow on the geographic structure observed in traits under selection. We used sequence data from the mitochondrial DNA cytochrome oxidase I and II subunits to document and compare the geographic structures of the parasitoid Agathis thompsoni and its moth host Greya subalba. We also documented the geographic structure of G. enchrysa and compared it to the geographic structure of its parasitoid Agathis n. sp. The results demonstrated that parasitoids and their hosts may have incongruent patterns of geographic structure as assessed by molecular markers. As a consequence, the geographic scale at which the interaction evolves may be different for each species involved in the interaction. Depending on the interplay of selection and gene flow, there may not be a one-to-one correspondence of traits important in the interaction between parasitoids and their hosts at the level of local populations. The geographic structures of A. thompsoni and G. subalba and Agathis n. sp. and G. enchrysa provide further evidence of the potential importance of the formation of geographic mosaics in coevolving parasitoid-host interactions and evolving interactions in general.     

Microsatellite isolation and characterization in the beneficial parasitoid wasp Diaeretiella rapae (M’Intosh) (Hymenoptera: Braconidae: Aphidiinae)

The primary parasitoid wasp, Diaeretiella rapae (M’Intosh), is used as a biological control agent in insect pest management strategies to combat aphid pests, mainly on cruciferous plants. To investigate the spatial scales over which this beneficial parasitoid wasp moves within an agricultural landscape, we developed five polymorphic microsatellite markers. The number of alleles per locus ranged from 11 to 21. Cross‐species amplification indicated that these loci may be useful in some other members of the Aphidiinae.     

Host instar preference and functional response of Aphidius transcaspicus, a parasitoid of mealy aphids (Hyalopterus species)

Aphidius transcaspicus Telenga (Hymenoptera: Braconidae: Aphidiinae) has been selected for use as a biological control agent against Hyalopterus pruni Geoffroy (Hemiptera: Aphididae) in prune orchards in California. In this study we focus on the host instar preference and functional response of A. transcaspicus. Host instar preference was determined using laboratory arenas by allowing female parasitoids to attack aphids on field-collected leaves of Prunus domestica L. supporting a natural range of H. pruni instars. A relative preference index indicated that A. transcaspicus was 1.4 times more likely to attack intermediate instars (2–4) than 1st or adult instars. The functional response of A. transcaspicus to H. pruni density was determined on enclosed P. domestica shoots in a prune orchard. The functional response to host density was linear (Type I) at the lowest aphid densities, and the asymptotic number of aphids attacked at higher aphid densities was 64.3?±?6.7 aphids day^?1.     

Genetic analysis of an introduced biological control agent reveals temporal and geographic change,with little evidence of a host mediated shift

The evolution of introduced biological control agents is largely un-explored. Although much is theorized, there is little empirical evidence quantifying the evolutionary dynamics of a biocontrol agent after release into a new environment. In this study we use Diachasmimorpha tryoni, a purposefully introduced biocontrol agent of Ceratitis capitata, to model and quantify spatial, temporal, and host-related evolutionary patterns. This parasitoid has undergone a host shift in its introduced environment, Hawaii, to the gall forming weed biocontrol agent, Eutreta xanthochaeta, an interaction likely mediated by competition for C. capitata with the egg-larval parasitoid Fopius arisanus. To elucidate potential evolutionary patterns we analyzed microsatellites and sequence data extracted from Hawaii and Australian population clusters defined by Structure, in Genepop, Canoco, and IBDWS. Our analysis revealed structuring of Hawaiian D. tryoni populations as defined by significant historic influences related to temporal structure, geographic space, host guild, and augmentative releases. The host-shift parasitoids were not genetically distinct from other Hawaii populations. There were small changes in microsatellite DNA at the population level, but only between Australia and Hawaii populations, not at the host level. These results show that D. tryoni has not undergone host-mediated evolution since introduction to Hawaii, despite the fact that they have expanded their host range in Hawaii to include the gall-forming E. xanthochaeta. To our knowledge this is the first study to quantify genetic differentiation of a biological control agent over geographic space and time using contemporary and museum specimens.     

A prospective model for the phenology of Microctonus hyperodae (Hymenoptera: Braconidae), a potential biological control agent of argentine stem weevil in New Zealand

A predictive phenological model is described for the parasitoid Microctonus hyperodae, introduced to New Zealand as a potential biological control agent against Argentine stem weevil Listronotus bonariensis. The model is based on development/temperature relationships obtained from experiments on the parasitoid in quarantine prior to its release, allowing early predictions of its phenology in different parts of the target pest's New Zealand range. In particular the model was used to predict the number of parasitoid generations each year, the degree of temporal synchrony between parasitoid adults and the susceptible adult pest stage, the order of parasitism and reproduction in the pest's life cycle as a possible basis for a simplified, discrete host/parasitoid population model, and the likely significance of ecotypic differences in development and diapause characteristics of the parasitoid. These applications demonstrate the potential for simple models to help in climate matching of classical biological control agents and estimation of their interaction with pest dynamics, using data obtainable prior to their introduction and release. In addition the model proved useful as a decision aid during the release programme, by indicating the likely effects of unusual weather and the need or otherwise for further parasitoid releases.     

Effects of constant and fluctuating temperatures on development of the coconut hispine beetle,Brontispa longissima (Coleoptera: Chrysomelidae) and two species of parasitoid

Abstract

The larval parasitoid Asecodes hispinarum Bou?ek has been used to control the coconut hispine beetle, Brontispa longissima (Gestro) in Vietnam. Although A. hispinarum has succeeded in suppressing B. longissima in southern Vietnam, biocontrol has not succeeded in central Vietnam. One factor underlying this failure might be the high temperature during the hot season. In the present work, we examined whether A. hispinarum parasitises the beetles within the range of temperatures that occur during the hot season in central Vietnam and we evaluated the potential use of the pupal parasitoid Tetrastichus brontispae Ferrière. Development of the beetle and two parasitoids was investigated at two constant temperatures (28 and 30°C) and at fluctuating temperatures from 25 to 35°C (corresponding to temperatures during the hot season in central Vietnam). B. longissima reached adulthood in all the temperature treatments. More than 70% of A. hispinarum emerged from hosts at 28°C, but none emerged at 30°C or under fluctuating temperature conditions, indicating that A. hispinarum cannot be used as a biological control agent in central Vietnam. However, T. brontispae could reproduce the next generation at all temperatures. These results suggest that T. brontispae is a potential agent for continuous biological control of B. longissima in central Vietnam.     

The influence of temperature on size as an indicator of host quality for the development of a solitary koinobiont parasitoid

The influence of aphid size on the host quality assessment and progeny performance of aphidiine parasitoids was examined using the mealy plum aphid parasitoid, Aphidius transcaspicus Telenga (Hymenoptera: Braconidae) and the black bean aphid, Aphis fabae Scopoli (Homoptera: Aphididae), as a readily acceptable alternate host. Aphid size in relation to stage of development was manipulated by rearing synchronous aphid cohorts at either 15 or 30 °C. At 15 °C, 2nd instar aphids were approximately the same size as 4th instar aphids reared at 30 °C. Cohorts of 30 aphids from each instar, reared at each temperature, were exposed to parasitism by a single parasitoid female for a period of 5 h. Overall susceptibility to parasitism did not vary between aphid cohorts, but the parasitoid response to aphid size differed significantly between rearing temperatures for both progeny sex ratio (parent female assessment of host quality) and larval growth and development (host suitability for parasitoid development). For aphids reared at 15 °C, the proportion of female progeny and emerging adult size for the parasitoid increased linearly with aphid size at the time of attack, while development time remained constant. In contrast, for aphids reared at 30 °C, the proportion of female progeny, emerging adult size, and the development time of the parasitoid all declined with aphid size at the time of attack. The contrasting responses of the parasitoid to host size for aphids reared at the two temperatures suggest that host quality is only indirectly related to aphid size among aphidiine parasitoids. The possible effects of higher temperatures on nutritional stress, obligate endosymbionts, and future growth potential of the aphids are discussed as explanations for the variation in host quality for parasitoid development.     

Host selection and reproductive success of French and Moroccan populations of the parasitoid, Microctonus aethiopoides (Hymenoptera: Braconidae)

French and Moroccan populations of the parasitoid Microctonus aethiopoides Loan were studied in the laboratory for their host selection, mating behavior, and reproductive success. The French strain, collected on Hypera postica (Gyllenhal), although capable of parasitizing and producing viable offspring on Sitona weevils, preferred Hypera weevils, its known target host. The Moroccan strain, collected on Sitona discoideus Gyllenhal, exhibited host specificity for Sitona. A partial reproductive isolation was observed between the two strains. Moroccan females mated more frequently with French males than did French females with Moroccan males. The pre-copulation time for mating pairs of opposite strains was significantly longer than that for mating pairs of the same strain. There was no significant difference in copulation time nor in larval and pupal duration between French and Moroccan strains. In summary, the French and Moroccan strains of M. aethiopoides are clearly separable by biological, behavioral, and morphometric traits and the preferred host for Hypera is the French strain and Sitona for the Moroccan strain. Consequently, geographic location and host source become important when considering this parasitoid as a potential biological control agent.     

Laboratory rearing and biological parameters of the eulophid Pnigalio agraules,a parasitoid of Cameraria ohridella

Predators as well as parasitoids native to Europe accept the exotic horse chestnut leafminer, Cameraria ohridella Deschka and Dimi? 1986 (Lepidoptera: Gracillariidae), either as prey or as host. However, the influence of these antagonists on the populations of the pest insect is so far very low. Therefore, efforts to develop an integrated pest management system against C. ohridella should include methods which foster the natural enemy complex. In the present study we developed a laboratory rearing method and investigated several biological parameters of Pnigalio agraules (Walker 1839) (Hymenoptera: Eulophidae), a dominant species in the horse chestnut leafminer’s parasitoid complex in many European regions. This native parasitoid satisfies three basic requirements for successful use as a biocontrol agent with regard to C. ohridella. The parasitoid’s fecundity, longevity and the speed of juvenile development by far exceeds that of the leafminer, enabling the parasitoid population to increase faster than that of the host. Furthermore, our results show that the impact of this species on C. ohridella populations has been previously underestimated, because non‐reproductive killing (i.e. host‐feeding and host‐stinging) of the hosts, resulting in considerable larval mortality of the leafminer, has not been quantitatively assessed. However, naturally occurring parasitoid populations have negligible impact on C. ohridella populations. Further studies, including experimental releases of P. agraules, are necessary to understand the constraints limiting the parasitoid’s performance in the field and to assess the potential benefits of releases for the control of C. ohridella.     

Behavior of Native Species Arrhenatherum Elatius (Poaceae) and Sonchus Transcaspicus (Asteraceae) Exposed to a Heavy Metal-Polluted Field: Plant Metal Concentration,Phytotoxicity, and Detoxification Responses

The application of vegetation cover for the phytomanagement of heavy metal-polluted soils needs prior investigation on the suitability of plant species. In this study, behaviors of Arrhenatherum elatius and Sonchus transcaspicus, two native perennial grasses that currently grow in a mine tailing, were investigated through plant metal concentration, phytotoxicity and their detoxification responses. Both of the species accumulated Ni, Cu, Cd, Co, Mn, Pb, Cr, and Zn in shoots far below criterion concentration as a hyperaccumulators; thus, neither of them were found to be hyperaccumulators. A. elatius accumulated metals in roots and then in shoots, on the contrary, in S. transcaspicus metals were preferentially accumulated in shoots. Plants exposure to such metals resulted in oxidative stress in the considered organs as indicated by the changes in chlorophyll fluorescence, chlorophyll contents, malondialdehyde (MDA) levels and antioxidative enzyme activities. A. elatius seemed to be more affected by metal-induced oxidative stress than S. transcaspicus. Correspondingly, S. transcaspicus showed a greater capacity to adapt to metal-induced oxidative stress, depending on more effective antioxidative defense mechanisms to protect itself from oxidative damage. These findings allowed us to conclude that both of these plant species could be suitable for the phytostabilization of metal-polluted soils.     

Mass‐rearing optimization of the parasitoid Psyttalia lounsburyi for biological control of the olive fruit fly

The olive fruit fly, Bactrocera oleae (Tephritidae), is a direct pest of olives that has invaded the Mediterranean Region and California. Psyttalia lounsburyi (Braconidae), a larval parasitoid from Africa, has been approved for release in the USA as a classical biological agent. However, it has been difficult to rear the parasitoid in the laboratory because it is multivoltine, and the host develops only in fresh olives, which are not available for most of the year. A method to rear the parasitoid on the factitious host, Mediterranean fruit fly (Ceratitis capitata) was developed, but it was not very efficient for producing large numbers of parasitoids needed for release. We developed a number of ways to improve the efficiency of rearing, including the frequency and duration of exposure for oviposition, optimizing the density of adult parasitoids, host age, as well as methods to quickly standardize the number of larvae exposed and to count emerging adult parasitoids. We significantly improved the number of progeny produced per female and the sex ratio of progeny. Thanks to these improvements, we produced in 2017 over 119,000 adults and shipped over 53,900 for release in California.     

Behavioural differences between Aphidius ervi populations from two tritrophic systems are due to phenotypic plasticity

The Palaeoarctic parasitoid Aphidius ervi Haliday (Hymenoptera, Aphidiidae) parasitises legume aphids in its region of origin. In Chile, it parasitises both legume and cereal aphids. This special situation was studied at two levels: (i) the host searching behaviour of A. ervi from two different tritrophic systems (Acyrthosiphon pisum on alfalfa and Sitobion avenae on wheat) was investigated in dual choice tests in a wind tunnel between odours from both A. pisum-alfalfa host plant complex (HPC) and S. avenae-wheat HPC, and (ii) the genetic structure of A. ervi populations from both sources using molecular markers. Responses of A. ervi females to volatile olfactory cues emanating from A. pisum-alfalfa HPC and S. avenae-wheat HPC were significantly higher towards the HPC on which they were reared during the last generation before experimentation, regardless of the origin of the parasitoid. As previously described for this parasitoid species, oviposition experience was also of major relevance in the preferences of female parasitoids. On the other hand, variation in mitochondrial DNA segments and RAPD-PCR polymorphism using total DNA showed the absence of host-based population structure and a high genetic homogeneity between these A. ervi populations. These results reject the possible existence of different host-strains of this parasitoid in Chile.     

Host Suitability of Four Cereal Stem Borers (Lepidoptera: Crambidae, Noctuidae) for Different Geographic Populations of Cotesia sesamiae (Cameron) (Hymenoptera: Braconidae) in Kenya

This study focused on the suitability of four species of cereal stem borers for the development of five geographic populations of Cotesia sesamiae (Cameron). C. sesamiae, an indigenous larval parasitoid of gramineous stem borers, is widespread in Africa. Four stem borers, Chilo partellus (Swinhoe), Chilo orichalcociliellus Strand (Lepidoptera: Crambidae), Busseola fusca Fuller, and Sesamia calamistis Hampson (Lepidoptera: Noctuidae), were offered to C. sesamiae for oviposition. Parasitoid individuals originated from five locations in Kenya. Biological parameters such as developmental time, percentage parasitism, progeny production, mortality of immature parasitoids, and proportion of female progeny were compared across host species. The two populations from western Kenya developed well on B. fusca. However, populations from the coast and the Eastern Province could not successfully parasitize B. fusca. With the exception of B. fusca, the percentage of hosts successfully parasitized by the different C. sesamiae populations was not different. The size of the host appeared to be an important factor influencing the development and reproductive potential of the parasitoid. We conclude that the different parasitoid populations were adapted to location-specific characteristics. Parasitoid–host compatibility must be evaluated before release for better establishment and colonization.     

Predicting the impacts of climate change on the biological control of Plutella xylostella by Diadegma semiclausum

1. The parasitoid Diadegma semiclausum Hellén (Hymenoptera: Ichneumonidae) has been widely adopted as a biological control agent for Plutella xylostella L. (Lepidoptera: Plutellidae) over the last 80 years. Earlier studies have found differential responses to temperature between the host and its parasitoid and demonstrated the multiple ways in which the parasitoid is more susceptible to elevated temperatures.
2. Using data from experimental studies, the modelling package CLIMEX was used to investigate the suitability of current climates for the host and its parasitoid and the effects on their potential global geographical distributions. The study was then extended to investigate possible changes to these distributions that might result under different climate change scenarios by 2080. The models predict that the global distributions of both the host and parasitoid will be reduced. These changes will not be proportionate and many areas in tropical, sub-tropical, and temperate regions that are currently suitable for D. semiclausum are predicted to become unsuitable for the parasitoid, whereas retaining suitability for P. xylostella. The seasonal dynamics of both the host and parasitoid are also predicted to be significantly reshaped under climate change.
3. Analysis of associations between annual P. xylostella outbreaks and weather conditions in three provinces in China with field data collected between 1995 and 2017 indicated significant effects of temperature on P. xylostella outbreaks at the beginning of the peak season in warmer provinces where P. xylostella can overwinter; such associations were not found in the colder provinces where it is unable to survive harsh winters.

     

Determination of the optimal parasitoid‐to‐host ratio for efficient mass‐rearing of the parasitoid,Sclerodermus pupariae (Hymenoptera: Bethylidae)

Sclerodermus pupariae Yang et Yao (Hymenoptera: Bethylidae) is used as a potential biocontrol agent for several buprestid and cerambycid larvae. This study aimed to enhance the efficiency of mass‐rearing of this parasitoid by investigating the fitness gain of this bethylid wasp, including the proportion of successful parasitism and development, brood size, sex ratio, proportion of winged female offspring, body size and longevity of female offspring, under eight different maternal parasitoid density treatments using Thyestilla gebleri Faldermann as host in the laboratory. The results indicated that the foundress densities did not affect the parasitism or emergence rate of this parasitoid. Brood size of the parasitoids increased significantly when the number of maternal wasps ranged from one to four. However, further increases in foundress number did not affect the parasitoid brood size. The sex ratios of S. pupariae were always female‐biased. The proportions of male in the progeny colonies were <10% throughout all experimental treatments. The percentage of winged female progeny was not significantly influenced by the density of adult maternal parasitoids. Body sizes of parasitoids significantly declined with increasing maternal parasitoid densities. Although the parasitoid body size reduced when maternal wasp number was higher, it could be compromised by the relatively higher number of female offspring produced. Further, more than 70% of the parasitoids remained alive when they were stored at 12°C for four months throughout the experiments. These findings suggest that exposure of four female wasps to a single host larva would result in the highest fitness of S. pupariae. Our findings might provide a new approach to enhance the efficiency of mass‐rearing of this bethylid wasp.     

Preliminary evaluation of the parasitoid wasp,Collyria catoptron,as a potential biological control agent against the wheat stem sawfly,Cephus cinctus,in North America

The wheat stem sawfly, Cephus cinctus (Hymenoptera: Cephidae) is the major pest of wheat in the northern plains of North America, with biological control providing a potentially useful management tool. Foreign exploration by the USDA-ARS identified Collyria catoptron (Hymenoptera: Ichneumonidae), a parasitoid wasp of Cephus fumipennis in northern China, as a potential biological control agent. Here we carry out a preliminary evaluation of the potential suitability of C. catoptron, as a candidate agent against C. cinctus. Specifically we: (1) Quantify the spatio-temporal variation in parasitism rates on the native host from collection sites in northern China over six years, and (2) Assess whether C. catoptron will oviposit and complete development in the novel targeted host. Maximum parasitism of the native host in the native range was 38%, exceeding the theoretical threshold for successful biological control. Site occupancy levels were high (83%) suggesting good colonisation abilities. C. catoptron clearly recognised and attacked C. cinctus, with parasitism levels exceeding those observed on the co-evolved host C. fumipennis. However, we found no evidence that C. catoptron can complete development in C. cinctus; no adults were reared from this host in either year of the study. In contrast 50–60% of the parasitoids survived to emerge as adults in C. fumipennis. Thus, C. catoptron is unlikely to be a suitable agent against C. cinctus due to basic host incompatibility. Future work will be directed towards developing conservation biological control approaches using native parasitoid species already present in the USA.