Oviposition preferences in Grapholita molesta: the relative importance of visual and olfactory cues

The oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), is a major pest of fruit trees worldwide. Females oviposit on the young leaves of peach [Prunus persica (L.) Batsch (Rosaceae)] shoots at twilight, when light intensity changes markedly. Previous researches have shown that the oviposition response of this moth is guided particularly by visual and olfactory cues from host plants. However, the relative importance of visual and olfactory cues in their oviposition preference is largely unknown. As a crepuscular moth, how do choices change as light intensity drops rapidly from day to night? In the present study, through two‐choice behavioural experiments, the oviposition responses of G. molesta to visual and olfactory cues (alone or in combination) from peach shoots were tested at four light intensities ranging from daylight to starlight. Grapholita molesta showed similar oviposition responses to plant cues under 1 000, 100, 1 and 0.01 mW m^?2 illumination. Olfactory cues from peach shoots attracted significantly more oviposition than the negative control, whereas visual cues alone did not. Furthermore, olfactory cues were more attractive than visual cues, and no significant interaction was observed between the responses to the two cues. Our findings indicate that G. molesta females mainly rely on olfactory cues to recognize oviposition sites, regardless of differences in light intensity. These results do not provide evidence for attraction to visual cues, but the possibility that the brightness of leaves might be used to guide oviposition is discussed.     

Synergism of pheromone and host‐plant volatile blends in the attraction of Grapholita molesta males

Control of Grapholita molesta (Busck) (Lepidoptera: Tortricidae), a major pest of stone and pome fruits, is successfully achieved by mating disruption. Under these conditions, tools other than conventional pheromone dispensers are needed for flight monitoring. The objective of the present work was to determine whether plant volatiles synergize male G. molesta attraction to a suboptimal dose of synthetic sex pheromone. The plant blend (referred to as 5VB), a mixture of three green leaf volatiles [(Z)‐3‐hexenyl acetate, (Z)‐3‐hexenol, and (E)‐2‐hexenal] and two aromatics [benzaldehyde (BZA) and benzonitrile (BZN)], was added to the suboptimal pheromone dose (2 ng on filter paper) in log steps (up to 10 000× the pheromone dose) to test synergism of pheromone and plant blends. In addition, the effect of individual plant volatiles on male responses was investigated by adding to the suboptimal pheromone dose each of the four‐compound plant‐volatile blends, resulting from eliminating one volatile from the 5VB at a time, or each plant volatile alone. Flight behaviour and the time to reach the source were recorded. The 5VB alone was not attractive to G. molesta males, but at a ratio of 1:1 000 (Ph:5VB) or higher, the attractiveness of the suboptimal pheromone dose increased, to a level similar to that of the optimal pheromone dose (10 ng). All tested plant volatiles, except BZA, synergized the response to the pheromone when added individually, but only (Z)‐3‐hexenol and BZN did so to a level not significantly different from the Ph:5VB blend. Aromatics had a stronger effect than green leaf volatiles (GLVs), because their removal, but not the removal of GLVs, decreased landing responses. The addition of the 5VB decreased significantly the time males needed to reach the odour source. The observed enhanced male attraction to mixtures of pheromone and plant volatiles will facilitate the development of lures for G. molesta adult flight monitoring.     

Factors influencing adult male Grapholita molesta dispersal in commercial Malus and Prunus host crops

The oriental fruit moth (OFM), Grapholita molesta (Busck) (Lepidoptera: Tortricidae), has been an economic pest of apples (Malus spp.) and peaches (Prunus spp.; both Rosaceae) in the eastern USA since the 1930s. Successful management of G. molesta with insecticides requires careful timing of these products based on sex‐pheromone trap captures of male moths. In Pennsylvania, apple and peach orchards are often planted adjacently. Factors such as trap distance from release points, host crop, and wind were considered in relation to male moth movement within and between hosts. Our practical objective was to understand male G. molesta dispersal within and between these two hosts to achieve better management. Recapture of moths decreased over distance; the majority of moths were recaptured 50–100 m from the release point regardless of crop type. Male G. molesta released into commercial apple and peach orchards were most likely to remain in the crop of release and to not exhibit host shifts over the season, even after peach fruit were harvested. Our analyses of wind direction data with respect to recapture showed that the likelihood of recapture was similar regardless of the orientation of the traps relative to the mean wind direction at peak flight times. The apple and peach host crops appeared to affect male G. molesta dispersal to the extent that wind in orchards affects their perception of pheromone. Crop effects were most likely due to the traps closest to the release points being in the same crop as the release points.     

Behavioral analysis of learning and memory in Anastrepha fraterculus

We evaluate the influence of prior exposure to artificial substrate for oviposition on learning and memory in the fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae). Some females were previously exposed to artificial fruits made of water, agar, and blackberry [Rubus spec. (Rosaceae)] or guava [Psidium guajava L. (Myrtaceae)] pulp for 48 and 72 h. We also studied adult flies exposed for 72 h to essential oil of lemongrass [Cymbopogon citratus (DC) Stapf, Poaceae] and adult flies from larvae exposed to the oil. Control females were naive with respect to these experimental substrates. Prior experience with blackberry‐based artificial fruits resulted in an increase in the number of punctures and deposited eggs by A. fraterculus, and memory lasted for up to 72 h. On the other hand, fly behavior was independent of exposure to guava‐based substrate. Prior exposure of 1‐ or 15‐day‐old females to artificial substrate with lemongrass oil modified innate substrate selection behavior. The scent of lemongrass oil during the larval stage modified innate oviposition responses of adult A. fraterculus. The study shows that A. fraterculus females are able to learn and retain information through chemical stimuli released by both host (blackberry and guava) and non‐host (lemongrass) species, and they can use olfactory memory obtained during the larval stage to select oviposition sites.     

Dual pheromone dispenser for combined control of codling moth Cydia pomonella L. and oriental fruit moth Grapholita molesta (Busck) (Lep., Tortricidae) in pears

Codling moth (CM) and oriental fruit moth (OFM) are very important orchard pests worldwide, and particularly in Victoria, Australia, where both species damage pome fruit. Individually CM and OFM have been controlled successfully by pheromone‐mediated mating disruption, but treatment of pome fruit with full registered rates of two individual hand‐applied dispensers for CM and OFM could be uneconomical for growers. Field trials conducted over three seasons in plots sprayed with insecticides consistently demonstrated that dual Isomate C/OFM TT dispensers, designed to disrupt both CM and OFM, were as effective as Isomate CTT and Isomate OFM Rosso dispensers applied individually in pears for control of CM and OFM respectively. The dual‐ and individual‐species dispensers reduced moth catches and fruit damage to a similar degree. The results suggest that combined control of CM and OFM in pears by applying dual Isomate C/OFM TT dispensers at the full‐recommended rate of 500 dispensers per hectare will be more economical than use of individual species dispensers, because the price and application cost of dual dispenser is about half that for individual dispensers.     

Geostatistical characterization of the spatial distribution of Grapholita molesta and Anarsia lineatella males in an agricultural landscape

Abstract:  The results obtained in spatial analysis of pheromone trap catches of oriental fruit moth, Grapholita molesta (Busck) and Peach twig borer, Anarsia lineatella (Zeller), are reported. The studies were performed in the Molise region, central Italy, during 2002 and 2003. Local distributions of oriental fruit moth and peach twig borer males were considered in a heterogeneous landscape. The aim of the study was to determine the temporal and spatial variation in distribution and abundance of the insect pests inside small plots of fruit orchards (apple, kiwi, peach, pear and plum fruits) and outside (in field crops, irrigation channels, hedgerows and a river), and to evaluate the importance of the host plants in relation to the adult distributions. Results showed that the main 'hot spot' for both lepidopterous pests was in a stone fruit orchard in the northern zone of the study area; other infested areas were in stone orchards and, in the case of A. lineatella , also in plum orchards. The river seems to act as a barrier rather than an ecological corridor. The observed spatio-temporal distributions of G. molesta and A. lineatella differed and were determined mainly by the location of the most important host plants and by the tendency of males to move in the environment, i.e. inside and outside the breeding and mating sites. Adequate knowledge of these spatial processes should be considered as an essential prerequisite for integrated pest management programmes in a precision farming approach.     

Olfactory responses of Grapholita molesta to different developmental stages of immature fruit of peach,plum, and apple

The Oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), is known to shift its host use from stone fruits (the primary hosts) to pome fruits (secondary hosts) as the season progresses. Grapholita molesta recognizes potential hosts by semiochemicals from fruits, but few studies have addressed the olfactory responses of G. molesta to host fruits during the seasonal transition of the moth from stone to pome fruits. So, we investigated the olfactory responses of larvae and gravid females of G. molesta to different fruit stages (collected on May 31, June 11, and June 25) of immature peach, plum, and apple fruits, using both multiple and two choice tests. In the multiple choice tests, for fruits collected after June 11, larvae showed a significant preference for peach as their first choice, compared to plum or apple. The final choice rate (when larvae had stayed on the fruit for more than 1 h) of larvae was also highest for peach fruits collected on June 11 and June 25. In two choice tests (Y-tube test), the response rate and overall choice rate of larvae and mated ovipositing adult females were also highest on peaches collected on May 31 (larvae) and June 25 (adults), respectively. In conclusion, we found that both the adults and larvae of G. molesta were highly attracted to immature peach fruits in laboratory choice tests.     

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.     

Distribution of the oriental fruit moth Grapholita molesta Busck (Lep., Tortricidae) infestation on newly planted peaches before and during 2 years of mating disruption

Abstract:  Oriental fruit moth (OFM) Grapholita molesta Busck (Lep., Tortricidae) is one of the most important pests of commercial stone fruit orchards in the Goulburn-Murray Valley region of Victoria, Australia. OFM populations have been successfully controlled by the use of the mating disruption (MD) technique for many years, but damage to shoot tips and fruit has now started to increase. The most severe damage under MD is found at the edge of peach blocks, adjacent to the pear blocks under insecticide treatment. In 1997–98, OFM infestation levels were examined in a newly planted peach block surrounded by older peaches, pears, apples and pasture. The infestation distribution was followed up for four consecutive years. No treatments were used against OFM for the first 2 years in the newly planted peaches, but in years 3 and 4 the whole block was treated with MD. At the end of year 2, shoot tip damage was randomly distributed throughout the newly planted peach block with no 'edge effect'. After MD was applied in year 3, the damage was confined to the edges of the block adjacent to insecticide-sprayed apples and pears. No 'edge effect' was detected along the border with an older peach block treated with MD or on the border with pasture. Extending the MD treated area for 25–30 m into the neighbouring apples and pears in year 4 reduced the 'edge effect'.     

Importance of trap liner adhesive selection for male moth catch (Lepidoptera: Tortricidae) with bisexual attractants

Studies compared moth captures of three pests (Lepidoptera: Tortricidae) of apple, Malus domestica Borkhausen, in delta traps using removable liners coated with either a sticky gel (SG) or a hot‐melt pressure sensitive (HMPS) adhesive. Laboratory and field studies with Cydia pomonella (L.), Choristoneura rosaceana (Harris) and Grapholita molesta (Busck) demonstrated that traps with either liner, baited with a pair of virgin females catch males, but at significantly different levels. In the field, male moth captures in traps with the HMPS liner were significantly greater than in traps with the SG liner for C. rosaceana, and G. molesta; but not for C. pomonella. Similar results were observed in laboratory studies using flight tunnels. Additional studies demonstrated that this difference in moth captures between liners was not due to levels of female mortality, but instead was correlated with the occurrence of the female's ventral abdominal surface becoming stuck in the adhesive. Studies showed that a significantly greater proportion of females of all species had their ventral abdomen stuck in the SG than HPMS adhesive on day 1, but only G. molesta and C. rosaceana on day 3. In addition, the tackiness of the two adhesives affected moth movement on the liner with males and females of all species moving farther on liners with SG than HMPS adhesive. A greater proportion of female G. molesta and C. pomonella were either stuck supined or laterally on the SG than HMPS adhesive, and females in this position captured as many males as when prone and unstuck on the liner. Our studies demonstrate that adhesives can secondarily influence male moth captures on trap liners when used with bisexual attractants, and that adhesive type should be considered when developing action thresholds.     

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.     

Investigating the molecular mechanisms of learning and memory using Caenorhabditis elegans

Learning is an essential biological process for survival since it facilitates behavioural plasticity in response to environmental changes. This process is mediated by a wide variety of genes, mostly expressed in the nervous system. Many studies have extensively explored the molecular and cellular mechanisms underlying learning and memory. This review will focus on the advances gained through the study of the nematode Caenorhabditis elegans. C. elegans provides an excellent system to study learning because of its genetic tractability, in addition to its invariant, compact nervous system (~300 neurons) that is well-characterised at the structural level. Importantly, despite its compact nature, the nematode nervous system possesses a high level of conservation with mammalian systems. These features allow the study of genes within specific sensory-, inter- and motor neurons, facilitating the interrogation of signalling pathways that mediate learning via defined neural circuits. This review will detail how learning and memory can be studied in C. elegans through behavioural paradigms that target distinct sensory modalities. We will also summarise recent studies describing mechanisms through which key molecular and cellular pathways are proposed to affect associative and non-associative forms of learning.

     

Manoeuvres used by flying male oriental fruit moths to relocate a sex pheromone plume in an experimentally shifted wind-field

ABSTRACT. In a wind-field experimentally shifted in direction by 35d?, flying male Grapholita molesta (Busck) zigzagging upwind either maintained contact with a pheromone plume and followed it across during the shift or lost it and commenced casting at c. 90d? across the shifting windline to locate it eventually in its new position. Males accomplished both of these results by integrating the previously described systems of optomotor anemotaxis and self-steered counterturning, but with faster reaction-times to pheromone on and off than heretofore calculated for this species. We found no evidence that males following the plume across used chemotaxis as proposed for another species, Rather, the sawtoothed-shaped tracks were a result of the anemotactic and counterturning systems responding rapidly and reiteratively to each loss and gain of pheromone along the plume in the shifting wind. The response to an increase or decrease in pheromone concentration by males was to change their course angle to more upwind or more crosswind, respectively, on the very first reversal (within c. 0.15 s) after the concentration changed. Because males adjusted their airspeeds more slowly to changes in concentration, the groundspeeds along the more upwind-orientated legs were lower than those along cross-wind legs, contributing to the sawtoothed shape of tracks of plume-followers. The self-steered counterturning programme also reacted quickly to concentration changes, the reversal intervals tending to be shorter following each contact with pheromone than after each excursion into cleaner wind. Following casting after losing the plume, males relocating the pheromone plume exhibited an upwind ‘surge’ of narrow zigzagging flight because on the first leg in the plume they steered a course more directly upwind than on the previous leg and increased the frequency of counterturning to its highest value while maintaining the relatively high airspeed acquired while casting.     

Control of the Oriental Fruit Moth, Grapholita molesta, Using Entomopathogenic Nematodes in Laboratory and Fruit Bin Assays

The oriental fruit moth (OFM), Grapholita molesta (Busck), which is among the most important insect pests of peaches and nectarines, has developed resistance to a wide range of insecticides. We investigated the ability of the entomopathogenic nematodes (EPN) Steinernema carpocapsae (Weiser), S. feltiae (Filipjev), S. riobrave (Cabanillas et al.), and Heterorhabditis marelatus (Liu and Berry) to control OFM under laboratory and fruit bin conditions. At a dosage of 10 infective juveniles (IJ)/cm² in the laboratory, S. carpocapsae caused 63%, S. feltiae 87.8%, S. riobrave 75.6%, and H. marelatus 67.1% OFM mortality. All four nematode species caused significant OFM larval mortality in comparison to the nontreated controls. Steinernema feltiae was used for the bin assays due to the higher OFM mortality it caused than the other tested EPN species and to its ability to find OFM under cryptic environments. Diapausing cocooned OFM larvae in miniature fruit bins were susceptible to IJ of S. feltiae in infested corner supports and cardboard strips. Treatment of bins with suspensions of 10 or 25 S. feltiae IJ/ml water with wetting agent (Silwet L77) resulted in 33.3 to 59% and 77.7 to 81.6% OFM mortality in corner supports and cardboard strips, respectively. This paper presents new information on the use of EPN, specifically S. feltiae, as nonchemical means of OFM control.     

Gas chromatography column as an ambient‐temperature volatile trap

Open‐tube volatile traps have largely been shunned in favor of solid adsorbent containing traps for the collection of volatile pheromones and attractants. Solid adsorbents require large solvent rinses and glass capillaries can be difficult to maneuver for the collection of volatiles from small or hard‐to‐reach odor sources. A gas chromatograph (GC) column (DB‐1), an open‐tube glass capillary, and a SuperQ^®‐containing capillary were compared for their collection efficiencies from rubber septa and live calling insects. All three traps captured similar ratios of test compounds from septa at airflows >10 ml per min. Eluting analytes from a packed adsorbent, SuperQ, required at least 30× more solvent than was required to collect all the pheromone from the open‐tube glass capillaries, and the GC column enjoyed an additional three‐fold reduced solvent volume compared to the glass capillary. Thus, analytes could be eluted from the GC‐column trap and directly analyzed on GC without solvent evaporation. We placed glass wool ‘plugs’ in both GC columns and glass capillaries and found no volatiles in these plugs, indicating that breakthrough did not occur during 1‐h collections at 25 ml per min. We demonstrate here that at ambient laboratory temperatures, a DB‐1 GC column effectively collects Oriental fruit moth sex pheromone volatiles from a rubber septum and live pheromone‐releasing moths. Release ratios of pheromone from rubber septa are consistent with earlier reports from static air systems, whereas the release ratio of the (Z)‐8‐dodecenyl alcohol (Z8‐12:OH) from female Grapholita molesta Busck (Lepidoptera: Tortricidae) differed from published results and is likely due to different collection methods or moth‐strain origin.     

Three physical barriers with obvious effects on Carposina sasakii selection behavior to two dominant jujube tree volatiles

The peach fruit moth (PFM), Carposina sasakii Matsumura (Lepidoptera: Carposinidae), a severe pest of Ziziphus jujuba Miller fruit trees (jujube, Rhamnaceae) mainly grown in China, has increasingly hindered the development of the jujube industry in recent years. In order to provide a theoretical basis for a physical obstruction method, we evaluated the effect of three commonly sold physical barriers on PFM host‐searching behavior. Two principal volatiles generated by the jujube tree, α‐farnesene and cis‐3‐hexenyl acetate (leaf acetate), were used as references to detect electroantennography (EAG) response values of 2‐, 4‐, and 6‐day‐old PFM adults. The results indicated different EAG responses toward α‐farnesene and leaf acetate between different ages of PFM, and males responded differently than females. The EAG responses were stronger in 4‐day‐old males to leaf acetate and in 2‐day‐old females to α‐farnesene. The two‐choice behavioral Y‐tube olfactometer assay confirmed that PFM was strongly attracted to α‐farnesene and leaf acetate. We determined the EAG and behavioral responses of PFM after spraying three kinds of physical barriers: Liquid Film Fruit Bag, High Lipid Film, and Jujube Protection No. 1. We obtained strong inhibitory effects on PFM attraction to both volatile lures using these barriers, including negative taxis by both females and males to Liquid Film Fruit Bag and Jujube Protection No. 1. We can infer that Liquid Film Fruit Bag is the most promising potential physical barrier for PFM control, with Jujube Protection No. 1 ranking as second‐best.     

Pheromone release rate determines whether sexual communication of Oriental fruit moth is disrupted competitively vs. non‐competitively

Communicational disruption mechanisms for Oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), were determined using a suite of mathematical tools and graphical operations that enable differentiation between competitive attraction and non‐competitive mechanisms of disruption. Research was conducted in 20 field cages, each covering 12 mature apple trees. Commercial monitoring lures releasing Oriental fruit moth pheromone at a rate of 0.04 μg h^?1 and distributed at densities of 0, 1, 2, 4, 8, and 17 per cage were used to evaluate the effect of low‐releasing dispensers on the disruption of sexual communication. Graphical analyses revealed that near‐female‐equivalent pheromone dispensers disrupted Oriental fruit moth competitively. Commercial mating disruption dispensers releasing Oriental fruit moth pheromone at 60 μg h^?1 and deployed at 0, 4, 6, 10, 15, 20, and 30 per cage were used to evaluate the effect of high‐releasing dispensers on the disruption of sexual communication. Oriental fruit moth disruption shifted to a non‐competitive mechanism for high‐releasing dispensers. This is the first time such a shift in disruption mechanism has been demonstrated against a background of otherwise identical experimental conditions. Near‐female‐equivalent pheromone dispensers were also used to quantify the additive effect of an attract‐and‐remove control strategy compared with competitive mating disruption. We report a five‐fold reduction in Oriental fruit moth captures under attract‐and‐remove compared to mating disruption using near‐female‐equivalent dispensers. Surprisingly, release of female Oriental fruit moths into these large‐cage disruption studies had no measurable impact on male trapping.     

Recurrent loops: Incorporating prediction error and semantic/episodic theories into Drosophila associative memory models

In 2003, Martin Heisenberg et al. presented a model of how associative memories could be encoded and stored in the insect brain. This model was extremely influential in the Drosophila memory field, but did not incorporate several important mammalian concepts, including ideas of separate episodic and semantic types of memory and prediction error hypotheses. In addition, at that time, the concept of memory traces recurrently entering and exiting the mushroom bodies, brain areas where associative memories are formed and stored, was unknown. In this review, I present a simple updated model incorporating these ideas, which may be useful for future studies.