Specificity of olfactory responses in the tephritid fruit fly, Rhagoletis pomonella

Electroantennograms (EAG) were recorded from, and behavior observed of female apple maggot flies, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), in response to over 60 individual esters. For acetates through decanoates, 2 methylbutyrates, and isobutyrates, we tested homologous series of systematically altered chain lengths. Most of the compounds had been isolated from behaviorally active fractions derived from extracts of volatiles produced by host fruits of R. pomonella. For the acetates through hexanoates, maximum EAG amplitudes were elicited by esters with chain lengths of 9 carbons and for the heptanoates through nonanoates, by 10 carbon esters. Recovery time, or the rate at which the EAG trace returned to the baseline following maximum depolarization, was slowest for straight chain esters that were 9–11 carbons long. Branching of the chain by addition of a methyl group to either side of the ester resulted in a decline in amplitude and a faster recovery time. Compared to EAG results, only 5 esters (butyl and pentyl hexanoate; propyl and butyl heptanoate; propyl octanoate) were highly active in wind tunnel bioassays, demonstrating (1) the hazard of assigning significance to EAG-active compounds without accompanying behavioral data, but more importantly; (2) a high degree of olfactory specificity. Maximum behavioral response was contingent upon the following rules regarding size and structure of the molecule. The ester must (1) be a straight chain; (2) be 10–11 carbons in length; (3) have an acid portion of 6–8 carbons and an alcohol portion of 3–5 carbons. One of the active esters, butyl hexanoate, appears in significant concentrations in the headspace of host fruit and, because this ester elicits such a pronounced behavioral response, our results suggest that R. pomonella is adapted to perception of a compound that is typical of its hosts.

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Résumé Les réactions de R. pomonella à plus de 60 esters différents ont été enregistreées par électroantennogrammes (EAG) et par ovservation du comportement. Pour les acétates, avec les décanoates, les 2-méthylbutyrates et les iso butyrates, nous avons examiné des séries homologues de chaînes aux longueurs systématiquement altérées. La plupart de ces composés ont été isolés à partir des fractions actives sur le comportement, issues des extraits des substances volatiles des fruits des plantes hôtes de cette téphritidae. Pour les acétates, avec les hexanoates, les EAG aux plus grandes amplitudes ont été obtenus avec des esters dont la longueur des chaînes est de 9 carbones, et avec les heptanoates et les nonanoates pour les esters à 10 carbones. Le temps de récupération ou temps mis par l'EAG pour revenir à l'ordonnée de départ après dépolarisation maximum, a été plus lent pour les esters à chaînes droites avec 9 à 11 carbones. La ramification de la chaîne par addition d'un groupe méthyl de chaque côté de l'ester a provoqué une réduction de l'amplitude et une accélération de la récupération. 5 esters seuls provoquent une forte réaction dans le tunnel à vent (hexanoates butilique et pentylique, heptanoates propylique et butylique, octanoate propylique). Ceci montre (1) le risque qu'il y a à attribuer une signification aux substances provoquant une réaction en EAG, lorsqu'il n'y a pas d'observations comportementales parallèles, et, surtout; (2) le degré élevé de spécificité olfactive.La résponse comportementale optimale obéit aux règles suivantes concernant la taille et la structure de la molécule. L'ester doit avoir: (1) une chaîne linéaire; (2) 10–11 carbone de longueur; (3) une portion acide de 6 à 8 C et une portion alcool de 3 à 5 C. Le butyl hexanoate, l'un des esters actifs, présent en concentration suffisante près de fruits utilisés, provoque pour cette raison une réponse comportementale tellement marquée, que nos résultats suggèrent que R. pomonella est adapté à la perception des substances caractéristiques de leurs hôtes.

     

Genetic variation among apple and hawthorn host races of Rhagoletis pomonella across an ecological transition zone in the Mid-Western United States

In previous studies, we have shown that apple and hawthorn populations of Rhagoletis pomonella (Diptera: Tephritidae) represent partially reproductively isolated and genetically differentiated host races; a result consistent with predictions of sympatric speciation models. The geographic pattern of allozyme variation for these flies is complex, however, as inter-host differences are superimposed on latitudinal allele frequency clines within the races. In addition, pronounced allele frequency shifts exist among R. pomonella populations across three major ecological transition zones in the mid-western United States. This suggests that selection related to environmental heterogeneity is responsible for the allele frequency shifts, but does not rule out secondary contact as an alternative possibility. Resolution of this issue is important, because if secondary contact is involved, then we would have to reassess the relationship host race formation has with speciation in the R. pomonella group.Here, we present results from a detailed genetic analysis of fly populations spanning the deciduous/prairie transition zone near the border between the states of Wisconsin and Illinois. Allele frequencies for hawthorn populations within the zone formed spikes, rather than the expected steps, and these frequency peaks correlated with variation in local ambient temperature conditions. Ambient temperature, and not secondary contact, therefore appears to be an important determinant of the shape of R. pomonella allele frequency clines. Allele frequency heterogeneity was also observed among apple populations, but was less pronounced compared to that for hawthorn flies. This suggests that ambient temperature differentially affects the host races, possibly through differences in the fruiting phenologies of apple and hawthorn trees. Several pairs of linked loci displayed concordant allele frequency changes and were in disequilibrium among both apple and hawthorn populations along the Wisconsin/Illinois transect. Although we do not know the reason for the observed pattern of disequilibrium, site to site variation in levels of inter-host migration, coupled with selection, seem the most likely explanations. We conclude by discussing how host specific adaptations, such as those associated with ambient temperature, may interact with host recognition traits to drive the sympatric speciation process for R. pomonella group flies.     

Flight behaviour of tsetse flies in host odour plumes: the initial response to leaving or entering odour

ABSTRACT. Free-flying, wild male and female Glossina pallidipes Aust. and G. m. morsitans Westw. were video-recorded in the field in Zimbabwe as they entered or left the side of a host-odour plume in cross-wind flight, or as they overshot a source of host odour in upwind flight (camera 2.5 m up looking down at a 3 times 2.5 m field of view at ground level). 80% of cross-wind odour leavers turned sharply ( turns 95^o), but without regard to wind direction (overshooters behaved essentially the same except that nearly 100% turned). Many fewer flies entering a plume cross wind turned ( c . 60%), and when they did they made much smaller turns ( 58^o); these turns were, however, significantly biassed upwind ( c . 70%). All three classes of fly had similar groundspeeds ( 5.5–6.5 m s^_1) and angular velocities ( 350–400^o s^-1). Clear evidence was obtained of in-flight sensitivity to wind direction: significantly more flies entering odour turned upwind than downwind, and odour losers turning upwind made significantly larger turns than average. The main basis for the different sizes of turn was the different durations of the turning flight, rather than changes in angular velocity or speed. No evidence was found of flies landing after losing contact with odour.     

Orientation of tsetse flies to wind, within and outside host odour plumes in the field

Abstract Free-flying wild tsetse flies ( Glossina pallidipes Aust. and G. m. morsitans Westw.) were video recorded in Zimbabwe as they flew within an artificial host odour plume at 3, 7 or 15 m from the source, or in no odour, with and without a 0.75 m² vertical, black visual target present aligned with the wind. With no visual target present, flights in odour were strongly biased upwind, and in the absence of odour strongly biased downwind. With the target present, between 16% and 40% of the upwind approaching flies responded visually as they passed the target, by circling it, in proportion to the proximity of the source (taken to be proportional to the mean odour concentration). Crosswind approaching flies (for whom the target will have been visible for some metres away) circled more frequently (34–56%), but without obvious correlation with the odour concentration. Circling flies also responded orthokinetically, by slowing down as they passed the target. The departure directions relative to the wind of flies leaving the target were significantly affected by the odour concentration. At 3 m they left the target in all directions, except possibly avoiding due upwind. At 7 m they left with an obliquely upwind bias, but at 15 m and also in no odour, they left with a strong crosswind bias.     

Behaviour of tsetse flies (Glossina) in host odour plumes in the field

ABSTRACT. In Zimbabwe, studies were made of the responses of Glossina pallidipes Austen and G.morsitans morsitans Westwood to artificial host odour using an incomplete ring of electrocuting nets. In a plume of synthetic host odour tsetse flew generally upwind, with 50–60% flying within 35^o of due upwind. More than 80% of tsetse flew at < 50 cm above ground level. Upon losing contact with odour they executed a reverse turn within about 2 m, and upon regaining contact they turned upwind. There were no clear differences in the responses of G.m.morsitans and G.pallidipes. Using electrocuting nets lying horizontally on the ground it was found that tsetse landed in the vicinity of the odour source, the propensity to land being greater for G.pallidipes than for G.m, morsitans , greater for immature than mature flies, and greater for males than females.     

Intra- and interspecific competition and host race formation in the apple maggot fly,Rhagoletis pomonella (Diptera: Tephritidae)

Intra- and interspecific resource competition are potentially important factors affecting host plant use by phytophagous insects. In particular, escape from competitors could mediate a successful host shift by compensating for decreased feeding performance on a new plant. Here, we examine the question of host plant-dependent competition for apple (Malus pumila)- and hawthorn (Crataegus mollis)-infesting larvae of the apple maggot fly, Rhagoletis pomonella (Diptera: Tephritidae) at a field site near Grant, Michigan, USA. Interspecific competition from tortricid (Cydia pomonella, Grapholita prunivora, and Grapholita packardi) and agonoxenid (subfamily Blastodacninae) caterpillars and a curculionid weevil (Conotrachelus crataegi) was much stronger for R. pomonella larvae infesting the ancestral host hawthorn than the derived host apple. Egg to pupal survivorship was estimated as 52.8% for fly larvae infesting hawthorn fruit without caterpillars and weevils compared to only 27.3% for larvae in harthorns with interspecific insects. Survivorship was essentially the same between fly larvae infesting apples in the presence (44.8%) or absence (42.6%) of interspecific insects. Intraspecific competition among maggots was also stronger in hawthorns than apples. The order or time that a larva exited a hawthorn fruit was a significant determinant of its pupal mass, with earlier emerging larvae being heavier than later emerging larvae. This was not the case for larvae in apples, as the order or time that a larva exited an apple fruit had relatively little influence on its pupal mass. Our findings suggest that decreased performance related to host plant chemistry/nutrition may restrict host range expansion and race formation in R. pomonella to those plants where biotic/ecological factors (i.e. escape from competitors and parasitoids) adequately balance the survivorship equation. This balance permits stable fly populations to persist on novel plants, setting the stage for the evolution of host specialization under certain mitigating conditions (e.g. when mating is host specific and host-associated fitness trade-offs exist).     

It''s about time: the evidence for host plant-mediated selection in the apple maggot fly, Rhagoletis pomonella, and its implications for fitness trade-offs in phytophagous insects

The apple maggot fly, Rhagoletis pomonella, Walsh (Diptera: Tephritidae), provides a unique opportunity to address the issue of host-related fitness trade-offs for phytophagous insects. Rhagoletis pomonella has been controversial since the 1860's when Benjamin Walsh cited the fly's shift from hawthorn (Crataegus spp.) to apple (Malus pumila) as an example of an incipient sympatric speciation event. Allozyme and mark-release-recapture studies have subsequently confirmed the status of apple and hawthorn flies as partially reproductively isolated and genetically differentiated host races, the hypothesized initial stage in sympatric divergence. Here, we review the ecological and genetic evidence for host-plant mediated selection in R. pomonella. We reach the following three major conclusions: First, although developmental timing is not everything, it is a good deal of the story. Differences in the fruiting phenologies of apple and hawthorn trees exert different selection pressures on the diapause and eclosion time characteristics of the host races. In particular, the 3-week earlier mean fruiting phenology of apples in eastern North America appears to select for a slower rate of metabolism or deeper pupal diapause in apple than hawthorn flies. Second, host-related fitness trade-offs for R. pomonella may not be due to disruptive selection affecting any one specific life-history stage. Rather, it is the sum total of directional selection pressures acting across different life-stages that generates divergent selection on apple and hawthorn flies. For example, selection favors the alleles Me 100, Acon-2 95 and Mpi 37 (or linked genes) in the larval stage in both host races. However, these same alleles are disfavored in the pupal stage to follow, where they correlate with early adult eclosion, and by inference premature diapause termination. Because apple trees fruit an average of 3 weeks earlier than hawthorn trees, this counter-balancing selection is stronger on apple-fly pupae. The net result is that the balance of selective forces is different between apple and hawthorn flies, helping to maintain the genetic integrity of the host races in sympatry in the face of gene flow. Finally, natural R. pomonella populations harbor a good deal of genetic variation for development-related traits. This variation allows fly populations to rapidly respond to temporal vagaries in local environmental conditions across years, as well as to broad-scale geographic differences that exist across the range of the species. Perhaps most importantly, this variation gives R. pomonella the flexibility to explore and adapt to novel plants. Taken together, our results underscore how difficult it can be to document host plant-related fitness trade-offs for phytophagous insects due to the need to consider details of the entire life-cycle of a phytophagous insect. Our findings also show how reproductive isolation can arise as a by-product of host-associated adaptation in insects, a central theme for models of sympatric speciation via host shifts.     

Influence of protein hydrolysate baits on oviposition in the apple maggot, Rhagoletis pomonella

Ovipositional responses of apple maggot (AM), Rhagoletis pomonella (Walsh), females were studied in the laboratory on apples (var: Golden Delicious) treated with different rates of four protein hydrolysate baits in choice and no-choice tests. Protein hydrolysate baits at rates of 0.5 and 1% had no significant effect, but oviposition was greatly reduced at higher rates of 5 and 10%. Apple maggot females exposed to apples treated with protein hydrolysate baits at a rate of 10% made 41–71% fewer punctures and laid 41–73% fewer eggs than in untreated control. No oviposition activity was shown on apples treated with 25 and 100% Nulure®. In no-choice tests the AM females laid 75–96% fewer eggs in apples treated with 10 and 25% Nulure compared to controls and no oviposition occurred in apples treated with 100% Nulure. Apple maggot females arrived in similar numbers on apples treated with 10% Nulure and untreated apples, but only 5% of those arriving on Nulure-treated apples showed ovipositor boring with no egg deposition while 60% of females arriving on untreated apples showed ovipositor boring activity and laid an average of 2.5 eggs per apple. In another experiment, individual AM females displayed similar behavioral responses to 10% Nulure-treated apples; none of the 56 females tested on treated apples displayed ovipositor boring activity, but 59% of the females (N=56) tested on untreated apples displayed ovipositor boring within 5 min of their arrival. Ninetyeight percent of AM females stayed and fed on fruit surfaces for 5 min on Nulure-treated apples without ovipositor boring compared to only 2% on untreated apples. Of the females that arrived on untreated apples, 39% flew away within 5 min without ovipositor boring compared to only 2% of those that arrived on Nulure-treated apples. Results of these two behavioral experiments suggest that upon arrival on a protein bait-treated apple, an apparent change of behavior occurs in AM females and instead of attempting to oviposit, they attempt to feed on fruit surfaces resulting in reduced oviposition activity. These results indicate that the feeding and oviposition-related activities of AM females are probably mutually exclusive and that the feeding behavior preempts oviposition activities on host fruits treated with higher rates of protein hydrolysate baits.     

Upwind flight of tsetse (Glossina spp.) in response to natural and synthetic host odour in the field

Abstract. In Zimbabwe, studies were made of the flight responses of tsetse ( Glossina spp.) to synthetic and natural ox odour using arrangements of electric nets.Tsetse flying away from a target showed a significant upwind bias when a blend of carbon dioxide (2/1 min), acetone (500 mg/h), octenol (0.4 mg/h), 4-methylphenol (0.8 mg/h) and 3-n-propylphenol (0.1 ma) was dispensed 15 m upwind, with c. 35% flying upwind.Without carbon dioxide this percentage was significantly reduced to 15% which was not significantly different from that with no odour (8%).This pattern was not altered by reducing the doses of acetone, octenol and phenols by 10–100 times, to levels comparable to those produced by an ox.With natural ox odour or a synthetic equivalent of ox odour dispensed from a ventilated pit 8 m upwind of the target, c. 28% flew upwind.This was reduced significantly to 15% if carbon dioxide was removed.In studies using a 17 m line of nets arranged orthogonally across the prevailing wind line, c. 50% of the catch was caught on the downwind side in the absence of odour.This increased significantly to c.60% when acetone, octenol and phenols were dispensed 15 m upwind, with or without carbon dioxide.With a shorter line (9 m) or an incomplete one (16.5 m long with 5 times 1.5 m wide gaps along its length) there was no change in the proportion caught downwind.For all three lines, dispensing odour upwind increased the catch 2–5 times on both the up-and downwind sides of the nets.It is concluded that a stronger upwind response to host odour is elicited when carbon dioxide is present.It is suggested that in nature upwind flight is very imprecisely orientated, with tsetse making flights up and down an odour plume 'searching' for a host.     

The effect of host odour on the landing responses of tsetse flies (Glossina morsitans morsitans) in a wind tunnel with and without visual targets

Abstract The effect of artificial host odour on the landing responses of males of Glossina m.morsitans West, and on their reaction to visual targets has been investigated in a wind tunnel. Landing was induced in flies that traversed steep odour gradients as they flew upwind and downwind across the edge of an odour plume, irrespective of whether visual targets were present or not; the landing response could be elicited over a wide range of odourconcentrations. When targets were present such odour gradients also tended to increase the proportion of landing flies which alighted on or near the targets; and the bigger the target, or the hungrier the flies, the greater was the propensity for target landing. In air which was more uniformly permeated with odour, the propensity to land on targets was increased only at high odour concentration.     

Flight behaviour of the black bean aphid, Aphis fabae, and the cabbage aphid, Brevicoryne brassicae, in host and non-host plant odour

Abstract. Walking alate virginoparae of Brevicoryne brassicae (L.) and Aphis fabae Scopoli were presented with odours of steam-distilled extracts of the non-host plants summer savoury ( Satureja hortensis L.) and tansy ( Tanacetum vulgare L.) in an olfactometer. No effects of the extracts were observed on B. brassicae. However, A.fabae were repelled by summer savoury and tansy odour; both extracts also masked an attractant response to bean (host plant) odour. In a flight chamber, air permeated with odour from host or non-host plants was blown over flying alates of both species, with a green, plant-mimicking target presented once a minute. The incidence of targeted (host-responsive) flight of B.brassicae was not affected by odour from a growing host plant ( Brassica oleoracea ) or a non-host plant tansy. Host plant ( Vicia faba ) odour did not affect the initial rate of climb or the incidence of targeted flight of A.fabae , but when the bean odour was alternated with odour from non-host tansy plants a greater number of targeted flights occurred in the host plant odour. The volatile extracts of tansy and summer savoury were also presented to flying A.fabae. Aphids flying in air permeated with tansy odour at 450g plant equivalents initiated fewer targeted flights than when flying in clean air. However, no differences in flight behaviour were observed with summer savoury extract. In a horizontal wind tunnel the tansy extract at 90 g plant equivalents blown across the surface of yellow targets reduced the numbers of alate A.fabae landing. The results indicate that plant odours can affect flight and landing of aphids.     

The flight and landing of tsetse (Glossina) in response to components of host odour in the field

ABSTRACT. Studies were conducted in Zimbabwe of the responses of Glossina morsitans morsitans Westwood and Glossina pallidipes Austen to various host odours using either arrangements of electrocuting nets or visual observations. Tsetse flying upwind in a plume of carbon dioxide, acetone and octenol turned downwind upon flying into a plume of acetone or octenol, but did not turn upon flying into a plume of carbon dioxide. They also turned in response to a transient decline in odour concentration. Tsetse landed on the ground in the vicinity of a source of natural odour or artificial odour containing carbon dioxide but not at sources of acetone or octenol only. The proportion of female G.pallidipes caught at a source of natural odour (37%) was significantly different from that caught at a source of synthetic odour (17%). Resting tsetse stimulated by natural odour took off sooner than non-stimulated flies and had a strong upwind bias in the direction of take off. Tsetse stimulated with artificial odour did not take off sooner than non-stimulated flies. It is suggested that there is an unidentified components) of ox odour that activates resting tsetse.     

Tarsal contact chemoreceptor for the host marking pheromone of the cherry fruit fly, Rhagoletis cerasi: responses to natural and synthetic compounds

Abstract. The European cherry fruit fly (Rhagoletis cerasi L.; Diptera, Tephritidae) marks cherries (Prunus avium L.) after oviposition with a host marking pheromone (HMP). The marking trail prevents additional oviposition by the same or other females into the same fruit. On the ventral side of the tarsi of both sexes, contact-chemoreceptor sensilla were identified which contain a receptor cell selectively sensitive to HMP. The HMP receptors of males were slightly more sensitive than those of females, suggesting that the more general term ‘host-marking pheromone’ is more appropriate than the previously used ‘oviposition deterring pheromone (ODP)’. The four structural isomers of the HMP, N(15R, S(β-glucopyranosyl)-oxy-8RS-hydroxypalmitoyl)-taurine, and various derivatives were synthesized and tested in an electrophysiological bioassay. Both the 8R,15R and the 8S,15RS isomers of the HMP were equally active with a threshold of about 2 times 10^-10M, and were shown to be present in the female faeces in similar proportions. The two 15S HMP isomers were about 13 times less active. Testing synthetic derivatives of the HMP molecule revealed that the presence of the four moieties of the molecule are important for the activity: taurine, palmitic acid, C₍₈₎ hydroxyl group, and glucose (C₍₁₅₎). The chain length of the fatty acid, the hydroxyl group at C₍₈₎ and the position of glucose at C₍₁₅₎ also influenced the activity. Only minor loss of activity (factor 2) relative to the natural molecule was observed when the methyl group in the C₍₁₅₎ position was removed. The removal of the β-glycosidically linked glucose (replaced by a hydroxyl group) resulted in about a 4-fold loss of activity. The cation of the HMP molecule seemed to have no effect on its activity, whereas both low and high pH reduced it significantly. Based on these results, field experiments have been initiated to control oviposition by cherry fruit flies on cherries applying the 15-desmethyl-HMP derivative.     

Irrigated agriculture and freshwater wetlands: A struggle for coexistence in the western United States

This paper is a review of the major environmental problems associated with irrigated agriculture in the western United States. Freshwater wetlands are being contaminated by subsurface agricultural irrigation drainage in many locations. Historic freshwater inflows have been diverted for agricultural use, and remaining freshwater supplies are not sufficient to maintain these important natural areas once they are degraded by irrigation drainage. Migratory birds have been poisoned by drainwater contaminants on at least six national wildlife refuges; waterfowl populations are threatened in the Pacific and Central flyways. Revised water allocation policies and regulatory actions are probably necessary to correct existing damage and prevent future problems. The benefits of maintaining healthy wetlands should be used as a rationale for negotiating increases in freshwater supplies. Cost analyses that show the importance of wetlands in dollar values are critical to the success of these negotiations. The next few years will provide unique opportunities for wetland managers to use cost analyses to make changes in water allocation policies. Federally subsidized water has supported and expanded agriculture at the expense of native wetlands for over 100 years in the western United States. This trend must be reversed if these wetlands and their fish and wildlife populations are to survive.