Behaviour of flying oriental fruit moth males during approach to sex pheromone sources

Abstract. The pheromone-modulated upwind flight tracks of Grapholita molesta (Busck) males were video recorded as they approached a point-source of pheromone in a wind tunnel. The field of view of the video recording was divided longitudinally into 33 cm sections and the flight behaviour of the males in these sections was measured and compared as they approached from 233 cm to 50 cm downwind of the pheromone source. As the males approached the source, their mean ground speeds decreased. The mean values of their track angles increased with respect to due upwind (0), indicating movement more across the wind. These changes resulted mainly from the males decreasing their air speeds as they progressed up the plume toward the source. They did not change the average direction of their steering (course angle). Thus, the increase in track angles resulted from the males allowing themselves to drift more in the wind as they approached the odour source. The males also increased their average rate of counterturning as they approached the source. The net result of all these behavioural changes was a track that slowed and grew narrower, giving the impression that the males were 'homing-in' on the pheromone source as they approached. Causes of these systematic changes in behaviour are considered with respect to the known systematic changes in pheromone plume structure as the distance to the source decreases.     

Pheromone-mediated optomotor anemotaxis and altitude control exhibited by male oriental fruit moths in the field

Abstract. In the field over short grass, pheromone-stimulated oriental fruit moth males, Grapholita molesta (Busck), flying under high windspeeds tended to steer courses more into the wind and to increase their airspeeds compared with those flying in low windspeeds.Thus, optomotor anemotaxis enabled the males to steer relatively consistent upwind track angles and to maintain an upwind progress of between c. 50–100 cm/s despite variable wind velocities.Zigzagging flight tracks were observed at both 10 m and 3 m from the source, as were tracks with no apparent zigzags.Transitions from casting to upwind flight or vice-versa were observed.The durations of the intervals between reversals during both upwind zigzagging flight and casting were consistent with those observed in previous wind-tunnel experiments.The control of altitude was more precise during upwind zigzagging flight than during casting.In general, the side-to-side deviations in the tracks were greater than the up-and-down deviations, with both the side-to-side and vertical distances and their ratios being consistent with previous wind-tunnel studies of pheromone-mediated flight.One difference between the field and laboratory flight tracks was that males in the field exhibited much higher airspeeds than in the wind tunnel.Males occasionally were observed to progress downwind faster than the wind itself, and further analysis showed that they were steering a downwind course in pheromone-free air following exposure to pheromone, which is the first time this has been recorded in moths.We propose that such downwind flight may aid in the relocation of a pheromone plume that has been lost due to a wind-shift, by enabling the moth to catch up to the pheromone as it recedes straight downwind away from the source.     

Field and laboratory electroantennographic measurements of pheromone plume structure correlated with oriental fruit moth behaviour

Abstract. Peak-to-trough electroantennogram amplitudes (bursts), caused by the individual filaments of a plume of female pheromone, diminish as high-emission-rate sources are approached by male Grapholita molesta , and this reduction is correlated with in-flight arrestment (ceasing to advance upwind). These findings are consistent with the hypothesis that one cause of in-flight arrestment in response to high-concentration point sources is the attenuation of the peak-to- trough amplitudes close to the source. High burst frequency, high pheromone flux, or low levels of continuous neuronal activity all are less well correlated with arrestment. Rather, arrestment appears due to a reduction of chemosensory input to the CNS during flight up the plume, even though the actual molecular concentration continues to increase. In a laboratory wind tunnel, upwind flight initiation by more than 20% of males was elicited only by pheromone source concentrations evoking significant fluctuations in EAG amplitudes at downwind release points. The burst frequencies that evoked high levels of upwind flight initiation ranged from a mean of 0.4-2.2 bursts/s. Because a previous study revealed that flying male G. molesta change their course angle within 0.15 s of losing or contacting pheromone, these EAG burst frequencies indicate that during flight in a pheromone plume, many manoeuvres are probably made in response to contact with individual plume filaments. Thus, upwind flight tracks may be shaped by hundreds of steering reactions in response to encounters with individual pheromone filaments and pockets of clean air. Field-recorded EAGs reveal that burst amplitudes diminish from 3 to 30 m downwind of the source, whereas burst frequencies do not, averaging c. 1/s at 3, 10 and 30 m downwind.     

Optomotor anemotaxis polarizes self-steered zigzagging in flying moths

ABSTRACT. Experiments with oriental fruit moth males, Grapholita molesta (Buck), provide evidence that a pheromone plume in zero wind elicits an endogenous, self-steered programme of counterturning (zigzagging) flight, and that wind experienced in flight establishes the polarity of the counterturns; they become aligned so that displacement occurs toward the source, even after the wind is stopped. In zero wind, males located a pheromone source more frequently when they had experienced a wind after having already taken flight before the wind was stopped (46%) compared with those that took flight later and therefore only experienced wind while they were in contact with the ground (14%). Furthermore, males placed in a stationary pheromone plume in zero wind located the source, eventually, on 21% of occasions. The flight tracks of these males, as well as those having experienced a wind only while on the ground, often exhibited repetitive counterturns (zigzags) of c. 180–200. However, the counterturns meandered around the flight tunnel, the inter-reversal track angles having no consistent direction. Sometimes the males displaced down-tunnel in the stationary plume, sometimes up, eventually locating the source and performing a courtship display. The inter-reversal track angles of males counterturning in wind, on the other hand, displayed a consistent orientation of c. 60 to either side of the wind line, resulting in consistent upwind displacement toward the source. With no pheromone present, with or without wind, counterturns were not observed.     

A non-anemotactic mechanism used in pheromone source location by flying moths

ABSTRACT. Male oriental fruit moths, Grapholitha molesta (Busck) (Tortricidae), continue to zigzag along a pheromone plume to the source in zero wind, if they have started flight with wind on. If the pheromone source is removed and the plume is hence truncated, moths flying in zero wind out of the end of the plume into clean air increase the width of their reversals and the angles of the straight legs of the tracks so they are more directly across the former wind line. Such moths reach the source less often than do those flying along a continuous plume. The males continue to zigzag up a plume in zero wind, apparently by a combination of sequential sampling of concentration along their path and the performance of an internal, self-steered programme of track reversals (zigzags) whose frequency increases with concentration. Visual feedback may aid in the still-air performance of the zigzags. We propose that both the sequential sampling (longitudinal klinotaxis) and self-steered counter-turning programme also are used in wind as well; anemotaxis apparently polarizes the direction of the zigzags to result in upwind displacement, and the narrow zigzags caused by the higher concentration in the plume keep the male 'locked on' to the odour.     

An analysis of anemotactic flight in female moths stimulated by host odour and comparison with the males' response to sex pheromone

ABSTRACT. The flight pattern of mated female navel orangeworm moths, Amyelois transitella (Walker), responding to odour from potential larval hosts is zigzagging upwind flight. However, at times these moths are capable of flying nearly directly upwind towards the odour source (track angles near 0). This response indicates that these females are capable of very accurate anemotactic control of their heading or course angle, since small angular errors in this measure would translate into larger deviations from direct upwind flight. Males of this species exhibit flight patterns similar to those of females, including track angles clustered about 0 when flying upwind to a source of the female-produced pheromone, but under these experimental conditions they flew with a higher average airspeed than the females. When females lose contact with an odour plume they initiate a well-defined programme of cross-wind counterturning or casting, which may normally increase their chances of retrieving contact with that plume when the wind direction shifts. The resultant track angles of females increase significantly by 0.8 s after plume loss, indicating that the female has initiated changes in both her course angle and airspeed. By 1 s after plume loss the females' track angles are no longer unimodally distributed about 0, but are bimodally distributed about -90 and +90. Males responded more rapidly to the loss of a pheromone plume, demonstrating a significant change in track angle 0.4 s after plume loss. Overall, female and male A.transitella exhibited remarkably similar anemotactic flight manoeuvres during upwind flight to odour sources as well as after plume loss.     

A pulsed cloud of sex pheromone elicits upwind flight in male moths

ABSTRACT. Male oriental fruit moths do not fly upwind in a continuous uniform cloud of pheromone, but readily do so when the cloud is pulsed at 1 or 0.5/s or when a plume from a point source of pheromone is placed within the continuous cloud. It is suggested that males of moth species that require such fluctuating pheromone stimulation for upwind flight will normally receive it from a filamentous, point-source-produced plume. However, we hypothesize that upwind progress may cease close to the source due to excessively high emission rates or inappropriate blend ratios, when fluctuating sensory output becomes attenuated, despite higher actual molecular concentration fluctuations.     

Effects of intermittent and continuous pheromone stimulation on the flight behaviour of the oriental fruit moth, Grapholita molesta

ABSTRACT. When male oriental fruit moths, Grapholita molesta (Busck) (Tortricidae), casting in clean air entered an airstream permeated with pheromone their flight tracks changed immediately on initial contact with pheromone, but after a few seconds returned to casting as if in clean air. The degree of change in the flight track was directly related to the concentration of pheromone. Although little net uptunnel movement occurred in response to the continuous stimulation provided by a uniformly permeated airstream, when an intermittent stimulus provided by a point-source plume was superimposed onto the permeated airstream moths were able to 'lock on' and zigzag uptunnel in the plume. The percentage of moths doing so corresponded to the difference between the peak concentration within the plume and the background concentration of pheromone permeating the airstream. Moths also locked onto, and flew upwind along the pheromone-clean-air boundary formed along a pheromone-permeated side corridor. Because a similar response was observed along a horizontal edge between a pheromone-permeated floor corridor and clean air, we conclude that the intermittent stimulation at the edge perpetuated the narrow zigzagging response to pheromone.     

Mating disruption barriers in pome fruit for improved control of oriental fruit moth Grapholita molesta Busck (Lep., Tortricidae) in stone fruit under mating disruption

Oriental fruit moth (OFM) Grapholita molesta Busck (Lep., Tortricidae) is a very important pest in commercial orchards in Victoria, Australia. Pheromone‐mediated mating disruption (MD) applied in stone fruit orchards successfully controlled OFM populations for many years, but damage to shoot tips and fruit at the edge of peach blocks located adjacent to pear blocks under insecticide treatments has become problematic. To improve protection of stone fruit against edge damage and outbreaks of OFM, all orchards were treated with sex pheromone dispensers for MD on an area‐wide basis. Area‐wide MD treatment, including all pome and stone fruit orchards in a discrete area, successfully controlled edge infestations of OFM, but was expensive. To reduce the cost of OFM control, sex pheromone dispensers for MD were applied as barrier treatments to 54–60 m of neighbouring pears adjacent to peaches under MD. Detailed monitoring of the OFM population, shoot tip and fruit damage assessments indicated that application of MD barriers on pears during two consecutive seasons provided sufficient control of OFM on peaches. This MD barrier treatment was able to reduce the number of OFM caught in all experimental peach blocks, with damage to shoot tips and fruit giving similar results to MD treatment of the whole neighbouring pear block. Extending the MD treatment area for 54–60 m into the neighbouring pear block significantly reduced the edge damage in MD‐treated peaches in the first season and almost eliminated OFM damage in the second season.     

The effects of pheromone concentration on the flight behaviour of the oriental fruit moth, Grapholitha molesta

ABSTRACT. Male oriental fruit moths, Grapholitha molesta (Busck) (Tortricidae), flew at lower overall and net ground velocities when they flew toward higher concentration pheromone sources. Turning frequency was greater with increased pheromone concentration, while the distance of turns from the plume axis back towards the axis decreased. Turning magnitude and inter-reversal track angles remained constant at all concentrations tested. Concomitant with the changes in ground velocity but constant inter-reversal angles, were decreases in airspeed, decreases in the moths' course angles and increases in their drift angles. The significance of these changes is discussed in relation to their possible role in a longitudinal chemoklinotactic programme of turning operating in conjunction with anemotaxis to allow location of a pheromone source in wind.     

Pheromone-mediated upwind flight of male gypsy moths, Lymantria dispar, in a forest

Abstract Lymantria dispar L. males flying upwind in a pheromone plume in a forest were video-recorded at 2.5, 10 and 20 m from the source of pheromone. Males flew slower and steered more across the wind as they approached the source. In concert, their ground speed decreased and track angles increased. In contrast to these changes, their drift angles were fairly constant and the transverse component of image flow, above and/or below the moths eyes, showed almost no change. The inter-turn duration (time between sequential turns), a temporal aspect of the male flight manoeuvres, showed a consistent but relatively small increase as the distance from the source increased. The flight tracks narrowed as the males approached close (2.5 m) to the source. Because of unpredicted correlations between physical variables (i.e. temperature, wind velocity) and the distance from the source, we used principal components analysis to generate a set of completely independent variables. Greater than 90% of the variability in the data could be explained by four principal factors which corresponded well with known relationships in the flight manoeuvres. All four of these factors showed a significant regression against distance to the source. Although uncontrolled factors such as temperature and wind velocity may have contributed to changes in flight behaviour, recent data indicate that, in addition to concentration, certain temporal and spatial characteristics (i.e. burst period, burst return period) of plumes in wind vary systematically with distance from the source. We propose that L.dispar males might adjust their flight manoeuvres in response to these changes.     

Effect of varying proportions of the alcohol component on sex pheromone blend discrimination in male Oriental fruit moths

ABSTRACT. The flight response of individual male Oriental fruit moths, Grapholitha molesta (Busck), was observed in a sustained-flight tunnel to 100 blend–dosage combinations of the three sex pheromone components: (Z)- and (E)-8-dodecenyl acetate and (Z)-8-dodecen-l-ol (1, 3, 10, 30 and 100 μg of Z8-12: AC with, 2, 6, 10, 20% E and, 0, 3, 10, 30 or100% OH alcohol added). Complete flights to the source were observed only to blend combinations containing all three components. Males exhibited highest response levels to two dosages (3 and 10μg) of the natural 6% E blend and these levels were relatively unaffected by changes in the proportion of Z8-12: OH. Certain treatments surrounding the peak area also elicited high response levels compared to the 6% E treatments, but these were strongly dependent on the proportion of OH in the blend. Hierarchical cluster analysis was utilized to compare and group treatments that elicited similar levels of response over the entire flight sequence and to derive an area of blend-dosage combinations within the matrix tested that elicited peak levels of attraction. Analysis of the response patterns for suboptimal treatments adjacent to the area of optimal attraction showed that response specificity was controlled by two major effects on flight behaviour, one occurring early in the flight sequence affecting male orientation to the odour plume, and the other occurring later in the sequence as an arrestment of upwind flight. These effects were strongly influenced by changes in the OH component and the E isomer, with the latter playing the critical role in effecting flight behaviour. Temporal analysis of the flight response to treatments within the optimal area showed that whereas the % OH did not significantly affect the magnitude of response, increasing the level of Z8-12: OH in the blend did significantly increase the duration of each phase of the behavioural sequence. Considering both the magnitude and temporal aspects of male response, optimal attraction in male OFM was best characterized by a small area of treatments around the peak 6% E blends that contained 10% OH. Finally, field tests showed a high degree of correlation between trends in response with changing proportion of Z8-12: OH as observed in the flight tunnel. Peak dosages were generally higher in the field, however, compared to those in the flight tunnel.     

Structured populations of the oriental fruit moth in an agricultural ecosystem

Intercontinental trade has led to multiple introductions of invasive pest species at a global scale. Molecular analyses of the structure of populations support the understanding of ecological strategies and evolutionary patterns that promote successful biological invasions. The oriental fruit moth, Grapholita (=Cydia) molesta, is a cosmopolitan and economically destructive pest of stone and pome fruits, expanding its distribution range concomitantly with global climate warming. We used ten newly developed polymorphic microsatellite markers to examine the genetic structure of G. molesta populations in an agricultural ecosystem in the Emilia‐Romagna region of northern Italy. Larvae collected in eight sampling sites were assigned to a mosaic of five populations with significant intra‐regional structure. Inferred measures of gene flow within populations implicated both active dispersal, and passive dispersal associated with accidental anthropogenic displacements. Small effective population sizes, coupled with high inbreeding levels, highlighted the effect of orchard management practices on the observed patterns of genetic variation within the sampling sites. Isolation by distance did not appear to play a major role at the spatial scale considered. Our results provide new insights into the population genetics and dynamics of an invasive pest species at a regional scale.     

The effects of different blend ratios and temperature on the active space of the Oriental fruit moth sex pheromone

Abstract The wing-fanning activation response of male Oriental fruit moths (OFM), Grapholita molesta (Busck), in the field to the three-component pheromone containing the female-produced ratio of components (Z8-12:OAc + 6% E8-12:OAc + 3% Z8-12:OH) was compared with the response to blends containing 2,10 and 20% E with 3% OH, and the 6% E blend containing 30 and 100% OH. Comparisons were made over three temperature ranges: 15–17, 20–21 and 26–28^oC. Both the maximum response distance and male response specificity were significantly altered by changes in odour quality as well as temperature. For blends containing different Z/E ratios the maximum response distance increased significantly with temperature. Response specificity was most pronounced at the 20–21^oC range, with males displaying a lower threshold for the natural 6% E ratio, evidenced by the fact that fewer males responded and at closer distances to the source with off-ratios. At 26–28^oC response specificity for the Z/E ratios was much reduced, primarily due to more males activating to off-ratios. With blends containing different proportions of Z8-12:OH in the 6% E blend, increasing temperature increased the maximum response distance for all treatments, but in addition increasing the proportion of OH alone from 3% to 30% significantly increased the maximum response distance over the three temperature ranges tested. This increase occurred without affecting the proportion of responders or the distribution of response distances around the mean value. However, with 100% OH added to the blend, whereas male response was high at 20–21^oC, the distribution of response distances was significantly more variable than with 3% or 30%, and male response was eliminated or very low at 15–17^oC and 26–28^oC. Our results support previous studies showing that peak response levels in this species are dependent on male perception of the natural blend of components, and that males have a high degree of specificity for the qualitative properties of the pheromone. However, the present results also extend those of previous flight tunnel tests in which response specificity was most pronounced in the upwind flight phase of the sequence, by showing that male OFM also display a     

Reduction of the response to sex pheromone in the oriental fruit moth,Grapholita molesta (Lepidoptera: Tortricidae) following successive pheromonal exposures

The effects of prior pheromonal experience upon the pheromone- mediated upwind flight response was examined in the oriental fruit moth, Grapholita molesta(Busck) (Lepidoptera: Tortricidae). Adult male G. molestawere subjected to a parallel series of staggered and repeated pheromonal exposures in a sustained-flight wind tunnel. Levels of response to pheromone in male G. molestasignificantly decreased in a (a) rectilinear function with increased ages of individuals, (b) logarithmic function of successive trials, and (c) steeper logarithmic function of successive trials with increased dosage of sex pheromone. The baseline levels of responding were not affected by either the (a) dosage of sex pheromone, (b) posteclosion ages of individuals for their initial exposures once the main effect of age itself was estimated, (c) elapsed time in hours between trials, or (d) discrete days of testing as integral intervals, disregarding hours within days.     

Temperature modulation of behavioural thresholds controlling male moth sex pheromone response specificity

ABSTRACT. The response specificity of male Oriental fruit moths, Grapholita molesta (Busck) and pink bollworm moths, Pectinophora gossypiella (Saunders), to different blends and doses of pheromone is altered dramatically by temperature. When acclimated and tested in a flight tunnel at 20^oC males of both species exhibit a high degree of specificity, with peak response levels occurring to a narrow range of blend-dose combinations close to the natural blend. When tested at 26^oC, however, males exhibit a significantly lower degree of specificity, with peak response occurring to a broader range of treatments. The change in response specificity results from shifts in behavioural threshold effects influencing plume orientation and initiation of upwind flight, as well as from arrestment of upwind flight, occurring later in the flight sequence. The observed changes in male behaviour suggest that the effect of temperature is directly on neural pathways involved in the perception of odour, and not simply the result of an increase in motor activity or a significant change in the release rate of the pheromone. The results support the threshold hypothesis for pheromone perception (Roelofs, 1978) as a general principle in the Lepidoptera, but also show that the degree of response specificity can be significantly affected by temperature.     

Volatile compounds from young peach shoots attract males of oriental fruit moth in the field

Abstract

The oriental fruit moth (OFM) is one of the most serious pests of commercial fruit orchards worldwide. Newly planted peach trees in particular, can be very attractive for mated OFM females for oviposition. Samples of airborne host plant volatiles from intact young peach shoot tips and old leaves of the same potted plants were collected and analyzed with a gas chromatograph-mass spectrometer. Chemicals present in young shoot tips, but not in old leaves, were used for field trials. Moth capture by traps with the synthetic chemicals was compared to that of the standard terpinyl acetate (TA) food trap. The TA food trap caught OFM males and mated females, but tested synthetic chemicals trapped only OFM males. We observed that the mixture of (Z)-3-hexenyl acetate: (E)-β-ocimene: (E)-β-farnesene in proportion 1:2:2 attracted OFM males. Further, 1 mg of (E)-β-ocimene, and that of (E)-β-farnesene also attracted OFM males.     

Sublethal effects of neuroactive compounds on pheromone response thresholds in male oriental fruit moths

The pheromone-mediated flight behavior of male Oriental fruit moths in a sustained-flight tunnel was observed after males were treated topically with sublethal concentrations of permethrin, carbaryl, chlordimeform, dieldrin, octopamine, serotonin, yohimbine, and cyproheptadine. With the exception of serotonin all compounds were found to disrupt one or more specific elements of the male precopulatory flight sequence. Among the insecticides, dieldrin was least active, whereas permethrin, carbaryl, and chlordimeform induced unique effects at specific phases of the sequence. Octopamine induced a hypersensitivity to the olfactory signal and mimicked one of the effects observed with chlordimeform. Yohimbine and cyproheptadine significantly decreased moth activation to the chemical signal but did not alter flight performance in responding moths. Yohimbine and cyproheptadine also reversed the effects induced by octopamine. The results of our study show that the complex precopulatory sequence of behaviors exhibited by males is very sensitive to sublethal concentrations of a range of neuroactive compounds.     

The analysis of flight paths of male Egyptian cotton leafworm moths, Spodoptera littoralis, to a sex pheromone source in the field

ABSTRACT. The flight of male Spodoptera littoralis (Boisd.) (Noctuidae) towards a pheromone source was recorded during the early part of the night using a cine camera and an image intensifier. The cine films were analysed frame by frame to produce flight tracks from which it was possible to calculate the mean advance rate of moths towards the pheromone source and their projected ground speed, for a series of positions downwind of the source. As wind speed was measured the moth's air speed was also estimated. The moths compensated for changes in wind speed by varying their air speed, hence maintaining a ground speed independent of wind speed. The ground speed itself was found to decrease as moths flew closer to the pheromone source.     

Close-range behaviour of male pea moths, Cydia nigricana, responding to sex pheromone re-released via the substrate

The close-range orientation behaviour of male pea moths, Cydia nigricana (F.) (Lepidoptera: Tortricidae), to a substrate contaminated with synthetic sex pheromone was studied in the field. The substrate was a sheet of polypropylene grass on a 1 m diam arena, on the upwind edge of which a pheromone trap had been placed previously for 1 h. After removal of the trap, moths continued to be attracted to the arena and to the vacated trap position. Video-recorded tracks showed that the moths behaved anemotactically whilst on the arena. When the arena was revolved so that the vacated trap position was on the downwind edge, moths continued to fly upwind on the arena, even though they were flying away from the original trap position and down a concentration gradient.