Genetic isolation between two sympatric host plant races of the European corn borer, Ostrinia nubilalis Hubner. II: assortative mating and host-plant preferences for oviposition

The European corn borer, Ostrinia nubilalis Hubner, colonized maize (Zea mays L.) after its introduction into Europe about 500 years ago and is now considered one of the main pests of this crop. In northern France, two sympatric host races have been described: one feeding on maize and the other on mugwort (Artemisia vulgaris L.) and hop (Humulus lupulus L.). In a previous study, we showed that mating between the two races may be impeded by differences in the timing of moth emergence and in the composition of the sex pheromone produced by the females. In this study, we further investigated the genetic isolation of these two races using strains from the maize (Z strain) and mugwort (E strain) races selected for diagnostic alleles at two allozyme loci. In a cage containing maize and mugwort plants and located in natural conditions, mating between individuals of the same strain occurred more often than mating between males and females of the E and Z strains. In particular, we obtained no evidence for crosses between Z females and E males. We also found that females of the Z strain laid their eggs almost exclusively on maize, whereas females of the E strain laid their eggs preferentially, but not exclusively, on mugwort. These results suggest that the genetic differentiation between the two host races may also be favored by host-plant preference, one of the first steps toward sympatric speciation.     

Genetic isolation between two sympatric host-plant races of the European corn borer,Ostrinia nubilalis Hübner. I. Sex pheromone,moth emergence timing,and parasitism

Adaptation to different environments may be a powerful source of genetic differentiation between populations. The biological traits selected in each environment can pleiotropically induce assortative mating between individuals of these genetically differentiated populations. This situation may facilitate sympatric speciation. Successful host shifts in phytophagous insects provide some of the best evidence for the ecological speciation that occurs, or has occurred, in sympatry. The European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae), colonized maize after its introduction into Europe by humans about 500 years ago. In northern France, two sympatric host races feed on maize (Zea mays) and mugwort (Artemisia vulgaris), respectively. We investigated the factors involved in the genetic isolation of these two races at a field site near Paris, France. We identified two biological differences that might make a significant contribution to the genetic divergence between sympatric populations feeding on the two host plants. First, assortative mating may be due to differences in the moth emergence pattern between the two races: mugwort-race moths emerged on average 10 days earlier than maize-race moths. In addition, the males emerged earlier than females in both races. Hence, the likelihood of mating between maize-race males and mugwort-race females was higher than that of mating between mugwort-race males and maize-race females. Second, the females feeding on mugwort and maize produced sex pheromones with different E/Z isomeric ratios of delta-11-tetradecenyl acetate. This difference in mate recognition systems reinforces the potential for assortative mating in the two races. During the experiment, overwintering mortality was much lower on maize than on mugwort. This difference was due to a braconid parasitoid wasp, Macrocentrus cingulum, that killed more than 50% of the larvae overwintering on mugwort but did not infest larvae diapausing on maize. Hence, by colonizing maize, European corn borer populations probably escaped from numerous predators, competitors, and parasitoids, such as M. cingulum. This decrease in host-associated selection may have favored the colonization of this new host. Finally, throughout this experiment we observed selection at two allozyme loci (or at linked loci): Tpi and Mpi. The Tpi locus is tightly linked with the genes involved in the response of the male to the sex pheromone and in developmental timing. The location of these traits on the Z chromosome may play a role in shortening the time required for the evolution of premating barriers.     

Sympatric host races of the European corn borer: adaptation to host plants and hybrid performance

The European corn borer (ECB), Ostrinia nubilalis, is a major pest of maize crops. In Europe, two sympatric host races are found: one feeds on maize (Zea mays) and the other mainly on mugwort (Artemisia vulgaris). The two host races are genetically differentiated, seldom crossing in the laboratory or in the field, and females preferentially lay eggs on their native host species. We conducted two independent experiments, in field and greenhouse conditions, to determine whether the two host races are locally adapted to their host species. The effect of larval density and the performance of hybrids were also investigated. Despite some differences in overall larval feeding performance, both experiments revealed consistent patterns of local adaptation for survival and for larval weight in males. In females the same trend was observed but with weaker statistical support. F1 hybrids did not seem to be disadvantaged compared with the two parental races. Overall, our results showed that both host races are physiologically adapted to their native host. The fitness trade-off between the two host plants provides a potential driving force for ecological speciation in this species.     

Evaluation of pheromone-baited traps for winter moth and Bruce spanworm (Lepidoptera: Geometridae)

We tested different pheromone-baited traps for surveying winter moth, Operophtera brumata (L.) (Lepidoptera: Geometridae), populations in eastern North America. We compared male catch at Pherocon 1C sticky traps with various large capacity traps and showed that Universal Moth traps with white bottoms caught more winter moths than any other trap type. We ran the experiment on Cape Cod, MA, where we caught only winter moth, and in western Massachusetts, where we caught only Bruce spanworm, Operophtera bruceata (Hulst) (Lepidoptera: Geometridae), a congener of winter moth native to North America that uses the same pheromone compound [(Z,Z,Z)-1,3,6,9-nonadecatetraene] and is difficult to distinguish from adult male winter moths. With Bruce spanworm, the Pherocon 1C sticky traps caught by far the most moths. We tested an isomer of the pheromone [(E,Z,Z)-1,3,6,9-nonadecatetraene] that previous work had suggested would inhibit captures of Bruce spanworm but not winter moths. We found that the different doses and placements of the isomer suppressed captures of both species to a similar degree. We are thus doubtful that we can use the isomer to trap winter moths without also catching Bruce spanworm. Pheromone-baited survey traps will catch both species.     

Small-scale dispersal of the European corn borer and its relevance for resistance management in Bt maize

The small-scale dispersal of the European corn borer (ECB) was studied in a release–recapture experiment using reared dye-marked adults. Thereby, six light trap cages were set up across two maize fields at 50-m intervals. In total, 736 marked ECBs were released, of which 10.2% were recaptured together with 212 unmarked naturally occurring adults after a period of 48 h. All marked–released individuals left the release point, with a mean dispersal distance of 195 m. Eighty-two per cent of the recaptured ECBs moved to the second maize field across a ditch and associated shrubs. The spatial and temporal patterns of incidence of naturally occurring ECBs in the traps were consistent with those of the marked moths and showed an inhomogeneous distribution. There was a highly significant relationship between male and female densities in the cages. No ECBs were caught during a period of adverse weather conditions. Dispersal distances may be influenced by plant size, weather conditions during the flight, pheromonal patterns in the field and the timing of the flight.     

Pheromone trap for the eastern tent caterpillar moth

The discovery that the eastern tent caterpillar Malacosoma americanum (F.) causes mare reproductive loss syndrome (MRLS), and thus has the potential to continue to result in major economic losses to the equine industry of Kentucky, has resulted in an intensive effort to identify practical means to monitor and control this defoliator, including these experiments to optimize a sex pheromone trap for this pest. A pheromone-baited delta trap with a large opening, such as InterceptST Delta, was more effective than other tested traps. Orange delta traps caught more moths than other tested colors. ETC males are caught at all tested heights within the tree canopy. For monitoring flights, setting traps at 1.5 m would allow easy counting of moths. A 9:1 blend of (E,Z)-5,7-dodecadienal (ETC-Ald) and (E,Z)-5,7-dodecadienol (ETC-OH) was most effective in capturing males. Increasing loading doses of a 3:1 blend (Ald:OH) resulted in the capture of increasing numbers of moths, but a 9:1 blend was more effective than 3:1 blend even at a nine-fold lower loading rate. Pheromone-impregnated white septa caught more moths than gray septa at the same loading dose. The advantages and limitations of using pheromone traps for monitoring M. americanum are discussed.     

Host-plant diversity of the European corn borer Ostrinia nubilalis: what value for sustainable transgenic insecticidal Bt maize?

The strategies proposed for delaying the development of resistance to the Bacillus thuringiensis toxins produced by transgenic maize require high levels of gene flow between individuals feeding on transgenic and refuge plants. The European corn borer Ostrinia nubilalis (Hübner) may be found on several host plants, which may act as natural refuges. The genetic variability of samples collected on sagebrush (Artemisia sp.), hop (Humulus lupulus L.) and maize (Zea mays L.) was studied by comparing the allozyme frequencies for six polymorphic loci. We found a high level of gene flow within and between samples collected on the same host plant. The level of gene flow between the sagebrush and hop insect samples appeared to be sufficiently high for these populations to be considered a single genetic panmictic unit. Conversely, the samples collected on maize were genetically different from those collected on sagebrush and hop. Three of the six loci considered displayed greater between-host-plant than within-host-plant differentiation in comparisons of the group of samples collected on sagebrush or hop with the group of samples collected on maize. This indicates that either there is genetic isolation of the insects feeding on maize or that there is host-plant divergent selection at these three loci or at linked loci. These results have important implications for the potential sustainability of transgenic insecticidal maize.     

Host‐associated divergence in sympatric host races of the leaf beetle Lochmaea capreae: implications for local adaptation and reproductive isolation

Ecological specialization is widely recognized as a major determinant of the emergence and maintenance of biodiversity. We studied two critical facets of specialization – local adaptation and habitat choice – in the host races of the leaf beetle Lochmaea capreae on willow and birch. Our results revealed that there is asymmetric disruptive selection for host use traits, and host races achieved different adaptive sets of life history traits through association with their host plant. Beetles from each host race exhibited food and oviposition preference for their own host plant. Reciprocal transplant displayed significant variation in host acceptance and performance: all families from the willow race rejected the alternative host plant before initiation of feeding and all died on this host plant. By contrast, all families from the birch race accepted willow for feeding, but they consumed less and performed less well. Intriguingly, families that performed well on birch also performed well on willow, suggesting positive genetic correlation rather than genetic trade‐offs. Our results suggest that the major proximal determinant of host specialization in the willow race is the behavioural acceptance of a plant rather than the toxicity of the food resource. However, in the birch race a combination of behavioural host acceptance and performance may play a role in specialization. Our study sheds light on the mechanisms by which divergent host adaptation might influence the evolution of reproductive isolation between herbivorous populations.     

Genetic structure of European and Mediterranean maize borer populations on several wild and cultivated host plants

Target pests may become resistant to Bacillus thuringiensis (Bt) toxins produced by trangenic maize (Zea mays L.). Untreated refuge areas are set aside to conserve high frequencies of susceptibility alleles: a delay in resistance evolution is expected if susceptible individuals from refuges mate randomly with resistant individuals from Bt fields. In principle, refuges can be toxin‐free maize or any other plant, provided it hosts sufficiently large pest populations mating randomly with populations from Bt‐maize fields. Our aim was to examine the suitability of several cultivated or weedy plants [pepper (Capsicum frutescens L.), sorghum (Sorghum spec.), sunflower (Helianthus annuus L.), cocklebur (Xanthium spec.), cantaloupe (Cucumis melo L.), and hop (Humulus lupulus L.)] as refuges for Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae), two major maize pests in southern Europe. Larvae of both species were collected on these plants. Their genetic population structure was examined at several allozyme loci. We found little or no evidence for an influence of geographic distance, but detected a significant host‐plant effect on the genetic differentiation for both species. Ostrinia nubilalis populations from sunflower, pepper, cocklebur, and sorghum appear to belong to the same genetic entity as populations collected on maize, but to differ from populations on hop. Accordingly, females from pepper and cocklebur produced exclusively the ‘Z’ type sexual pheromone, which, in France, characterizes populations developing on maize. Qualitatively, these plants (except hop) could thus serve as refuges for O. nubilalis; however, they may be of little use quantitatively as they were found much less infested than maize. Sesamia nonagrioides populations on maize and sorghum reached comparable densities, but a slight genetic differentiation was detected between both. The degree of assortative mating between populations feeding on both hosts must therefore be assessed before sorghum can be considered as a suitable refuge for this species.     

Molecular differentiation at nuclear loci in French host races of the European corn borer (Ostrinia nubilalis)

French populations of the European corn borer consist of two sympatric and genetically differentiated host races. As such, they are well suited to study processes that could be involved in sympatric speciation, but the initial conditions of host-race divergence need to be elucidated. Gene genealogies can provide insight into the processes involved in speciation. We used DNA sequences of four nuclear genes to (1) document the genetic structure of the two French host races previously delineated with allozyme markers, (2) find genes directly or indirectly involved in reproductive isolation between host races, and (3) estimate the time since divergence of the two taxa and see whether this estimate is compatible with this divergence being the result of a host shift onto maize after its introduction into Europe approximately 500 years ago. Gene genealogies revealed extensive shared polymorphism, but confirmed the previously observed genetic differentiation between the two host races. Significant departures from the predictions of neutral molecular evolution models were detected at three loci but were apparently unrelated to reproductive isolation between host races. Estimates of time since divergence between French host races varied from approximately 75,000 to approximately 150,000 years, suggesting that the two taxa diverged recently but probably long before the introduction of maize into Europe.     

桃蛀果蛾寄主生物型分化的RAPD分析

以山茱萸蛀果蛾Carposina coreana Kim为外群,用44个随机引物对苹果、山楂、桃、杏、枣和酸枣等6种寄主植物上的桃蛀果蛾Carposina sasakii Matsumura进行了RAPD 分析。结果表明,不同寄主上的桃蛀果蛾之间遗传差异出现了较大的分化,桃蛀果蛾6个寄主生物型之间的平均遗传距离在0.040～0.637之间,尤其是杏生物型分化最大。经聚类分析,可将不同寄主上的桃蛀果蛾明显分成两个类群,杏生物型为一群,其余为一群。两个类群间遗传距离为0.465,亚群内部的遗传距离为0.040～0.369。推测杏树上的“桃蛀果蛾 ”已经分化成为不同的物种。     

Effects of phenylacetaldehyde and (Z)-3-hexenyl acetate on male response to synthetic sex pheromone in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Abstract  The influence of two synthetic plant volatiles on the number of male Helicoverpa armigera (Hübner 1808) (Lepidoptera: Noctuidae) caught in sex pheromone-baited traps was studied in field plots in Australia. Phenylacetaldehyde and ( Z )-3-hexenyl acetate were combined with synthetic sex pheromone in traps. Generally, most males were captured in traps baited with pheromone alone and more males were captured by the mixture containing ( Z )-3 hexenyl acetate than that containing phenylacetaldehyde. At the three concentrations studied, neither volatile synergised the activity of the pheromone. Traps baited with the plant volatiles alone caught very few moths of either sex.     

Improved monitoring of female codling moth (Lepidoptera: Tortricidae) with pear ester plus acetic acid in sex pheromone-treated orchards

The performance of clear delta traps baited with 3.0 mg of pear ester, ethyl (E,Z)-2,4-decadienoate, and 5.0 ml of acetic acid in separate lures was compared with orange delta traps baited with a single lure containing 3.0 mg of both pear ester and the sex pheromone, (E,E)-8,10-dodecadien-1-ol (codlemone) for codling moth, Cydia pomonella (L.), in apple, Malus domestica (Borkhausen). Residual analyses and field tests demonstrated that both the pear ester and acetic acid lures were effective for at least 8 wk. The two trap-lure combinations caught a similar number of total moths in an orchard treated with sex pheromone dispensers during short-term trials in 2008. However, the mean catch of female moths was significantly higher and male moths significantly lower in clear traps baited with pear ester and acetic acid versus orange traps baited with pear ester and codlemone. Season-long studies were conducted with these two trap-lure combinations in orchards treated with (n = 6) and without (n = 7) sex pheromone dispensers during 2009. The two trap-lure combinations caught similar numbers of moths in dispenser-treated orchards. In contrast, total catch was significantly higher (>2-fold) in the orange compared with the clear traps in untreated orchards. The clear caught >6-fold more females than the orange trap in both types of orchards. These studies suggest that deploying clear delta traps baited with pear ester and acetic acid can be an effective monitoring tool for female codling moth and an alternative to codlemone-baited traps in sex pheromone-treated orchards.     

Field attraction of the stink bug Euschistus conspersus (Hemiptera: Pentatomidae) to synthetic pheromone-baited host plants.

The attraction of the stink bug Euschistus conspersus Uhler to sources of the synthetic pheromone component methyl (2E,4Z)-decadienoate was investigated in a series of field experiments in native vegetation surrounding commercial apple orchards in Washington. In experiments with pheromone lures placed inside two different tube-type traps, stink bugs were attracted to the immediate area around traps in large numbers, but very few were caught in the traps. Pheromone lures attached directly to the host plant mullein, Verbascum thapsus L., demonstrated that these 'baited" plants attracted significantly more E. conspersus than unbaited plants. Spring (reproductive) and summer (reproductively diapausing) E. conspersus adults, both males and females, were attracted to pheromone-baited plants. There was no significant difference in the number of male or female E. conspersus attracted to pheromone-baited traps or plants in any of the experiments, further characterizing methyl (2E,4Z)-decadienoate as an aggregation, and not a sex pheromone. Stink bug aggregations formed within 24-48 h of lure placement on mullein plants and remained constant until the lure was removed after which aggregations declined over 3-4 d to the level of unbaited plants. The implications of these studies for E. conspersus monitoring and management are discussed.     

Survival relative to new and ancestral host plants,phytoplasma infection,and genetic constitution in host races of a polyphagous insect disease vector

Dissemination of vectorborne diseases depends strongly on the vector's host range and the pathogen's reservoir range. Because vectors interact with pathogens, the direction and strength of a vector's host shift is vital for understanding epidemiology and is embedded in the framework of ecological specialization. This study investigates survival in host‐race evolution of a polyphagous insect disease vector, Hyalesthes obsoletus, whether survival is related to the direction of the host shift (from field bindweed to stinging nettle), the interaction with plant‐specific strains of obligate vectored pathogens/symbionts (stolbur phytoplasma), and whether survival is related to genetic differentiation between the host races. We used a twice repeated, identical nested experimental design to study survival of the vector on alternative hosts and relative to infection status. Survival was tested with Kaplan–Meier analyses, while genetic differentiation between vector populations was quantified with microsatellite allele frequencies. We found significant direct effects of host plant (reduced survival on wrong hosts) and sex (males survive longer than females) in both host races and relative effects of host (nettle animals more affected than bindweed animals) and sex (males more affected than females). Survival of bindweed animals was significantly higher on symptomatic than nonsymptomatic field bindweed, but in the second experiment only. Infection potentially had a positive effect on survival in nettle animals but due to low infection rates the results remain suggestive. Genetic differentiation was not related to survival. Greater negative plant‐transfer effect but no negative effect of stolbur in the derived host race suggests preadaptation to the new pathogen/symbiont strain before strong diversifying selection during the specialization process. Physiological maladaptation or failure to accept the ancestral plant will have similar consequences, namely positive assortative mating within host races and a reduction in the likelihood of oviposition on the alternative plant and thus the acquisition of alternative stolbur strains.     

Influence of host plant,geography and pheromone strain on genomic differentiation in sympatric populations of Ostrinia nubilalis

Patterns of mating for the European corn borer (Ostrinia nubilalis) moth depend in part on variation in sex‐pheromone blend. The ratio of (E)‐11‐ and (Z)‐11‐tetradecenyl acetate (E11‐ and Z11‐14:OAc) in the pheromone blend that females produce and males respond to differs between strains of O. nubilalis. Populations also vary in female oviposition preference for and larval performance on maize (C4) and nonmaize (C3) host plants. The relative contributions of sexual and ecological trait variation to the genetic structure of O. nubilalis remains unknown. Host‐plant use (¹³C/¹⁴C ratios) and genetic differentiation were estimated among sympatric E and Z pheromone strain O. nubilalis males collected in sex‐pheromone baited traps at 12 locations in Pennsylvania and New York between 2007 and 2010. Among genotypes at 65 single nucleotide polymorphism marker loci, variance at a position in the pheromone gland fatty acyl‐reductase (pgfar) gene at the locus responsible for determining female pheromone ratio (Pher) explained 64% of the total genetic differentiation between males attracted to different pheromones (male response, Resp), providing evidence of sexual inter‐selection at these unlinked loci. Principal coordinate, Bayesian clustering, and distance‐based redundancy analysis (dbRDA) demonstrate that host plant history or geography does not significantly contribute to population variation or differentiation among males. In contrast, these analyses indicate that pheromone response and pgfar‐defined strain contribute significantly to population genetic differentiation. This study suggests that behavioural divergence probably plays a larger role in driving genetic variation compared to host plant‐defined ecological adaptation.     

Phylogeography and host race differentiation in the European mistletoe (Viscum album L.)

Mistletoes are bird dispersed, hemi-parasitic shrubs infecting a large number of woody host plants. Chloroplast fragment length polymorphisms were used to study genetic differentiation among presumed host races, population genetic structure, and to elucidate the postglacial migration history of mistletoe ( Viscum album ) across the entire natural distribution range in Europe. The populations sampled belong to four closely related taxa, three of which are widely distributed and differ in their host trees, whereas a fourth taxon is rare and endemic to the Island of Crete. The molecular analysis of chloroplast DNA variation supported the distinction of these four taxa. We further found evidence for phylogeographical structure in each of the three widely distributed host races. Independent of host race, mistletoe haplotypes from Turkey were distinct and distant from those found elsewhere in Europe, suggesting that highly differentiated populations, and possibly new taxa, exist at the range limit of the species.     

Design and elevation of sex-attractant traps for pea moth, Cydia nigricana (Steph.) and the effect of plume shape on catches

• 1 Catches of male pea moths in six designs of trap, each containing (E,E)-8,10-dodecadienyl acetate as an attractant, were compared; triangular-shaped traps caught most moths.
• 2 The vertical distribution of moths within a pea crop and the size of catches at different levels varied in different wind speeds. Three-quarter crop height is probably the optimum height for monitoring.
• 3 Five trap designs (including the triangular type) captured only 20–30% of individuals landing on them; water traps were slightly more efficient.
• 4 There was an eight-fold range in the number of moths caught depending on the sticky material used. ‘Bird-Tanglefoot’ was most effective.
• 5 Smoke plumes, emitted from different trap designs in the field to simulate plumes of attractant, were photographed and measured. The effect of trap design on plume shape was confirmed in wind tunnel tests.
• 6 Field tests showed that the more elongated the mean plume emitted from a trap the greater the catch.
• 7 The tracks of individual moths flying to traps are discussed in relation to current views on orientation to odour sources.

     

Genetic races associated with the genera and sections of host species in the holoparasitic plant Cytinus (Cytinaceae) in the Western Mediterranean basin

* Speciation via race formation is an important evolutionary process in parasites, producing changes that favour their development on particular host species. Here, the holoparasitic plant Cytinus, which has diverse host species in the family Cistaceae, has been used to study the occurrence of such races. * Amplified fragment length polymorphism (AFLP) analyses were performed on 174 individuals of 22 populations parasitizing 10 Cistaceae species in the Western Mediterranean basin. * Neighbour-joining, multivariate ordination analyses, and individual-based Bayesian analyses, clustered Cytinus populations into five well-characterized genetic races that, overall, agreed with the taxonomic sections of their hosts. In the AMOVA, among-races differences accounted for almost 50% of the genetic variation. The isolation-by-distance model was not supported by a Mantel test among Cytinus populations (r = 0.012; P = 0.456). All races showed low within-population genetic diversity, probably as a result of restricted pollen flow aggravated by flowering asynchrony, restricted seed dispersion, or stochastic processes. * The genetic differentiation among the five races of Cytinus is congruent with the view that these races are well-characterized lineages that have evolved independently as a result of selective pressures imposed by their hosts. This pattern, with genetically distinctive groups associated with the infrageneric sections of the host species, has not been reported previously for parasitic angiosperms.