Variability in the efficacy of sex pheromone lures for monitoring oriental fruit moth (Lepidoptera: Tortricidae)

Studies were conducted in Chile and the United States to compare the attractiveness of various commercial sex pheromone lures and two experimental lures for oriental fruit moth, Grapholita molesta (Busck), in peach orchards treated with or without sex pheromone dispensers. The experimental lures contained the three‐component sex pheromone blend of G. molesta: Z‐8‐dodecenyl acetate, E‐8‐dodecenyl acetate and Z‐8‐dodecenol (Z8‐12:OH), and the sex pheromone of codling moth, Cydia pomonella (L.), (E,E)‐8,10‐dodecadien‐1‐ol, (codlemone). Commercial lures varied in their substrate, initial loading and blend ratio of components. Significant differences in male catches were found among commercial lures in orchards treated with or without sex pheromone dispensers. Experimental lures with the addition of codlemone significantly increased the catches of G. molesta using lures loaded with 0%, 1% or 5% Z8‐12:OH in the G. molesta blend compared with the same ratio of components in just the G. molesta blend. The experimental lures were significantly more attractive than all commercial lures in the untreated orchard. However, moth catch with the experimental lures in the sex pheromone‐treated orchard was only intermediate among all of the lures tested. These findings highlight the need to develop more effective and standardized lures that can be used in trap‐based monitoring programme for this important pest.     

Effects of different sex pheromone compositions and host plants on the mating behavior of two Grapholita species

The two congener species Grapholita molesta and Grapholita dimorpha share two major sex pheromone components: cis-8-dodecenyl acetate (Z8-12Ac) and trans-8-dodecenyl acetate (E8-12Ac). In fact, commercial sex pheromone lures composed of only these two major components attract the males of both species. In this study, we aimed to determine the reproductive isolation components of these two species by analyzing the effects of the minor sex pheromone components and host plants. First, different ratios of the two major sex pheromone components were greatly favored by either male species. Sex pheromone gland extracts of G. dimorpha contained a lesser proportion of Z8-12Ac than that of G. molesta. In the three (apple, pear, and peach) orchards investigated in this study, a larger number of G. molesta males were attracted to the 95:5 pheromone mixture (Z8-12Ac and E8-12Ac, respectively), while a larger number of G. dimorpha males were attracted to the 85:15 mixture. Second, there was a significant variation in male attractions in different host plants. G. molesta males were more attracted to the sex pheromone lure in the apple orchards than that in the pear and peach orchards. In contrast, G. dimorpha males were more attracted to the lures in the pear and peach orchards than that in the apple orchard. Third, the minor sex pheromone components were important for reproductive isolation. Among the four minor components tested, addition of (Z)-8-dodecenol (Z8-12OH) to the major sex pheromone components significantly suppressed male attraction in G. dimorpha and slightly elevated male attraction in G. molesta. The discriminating effect of Z8-12OH was further validated using male electroantennogram analysis. These results suggest that reproductive isolation between two congeners can be achieved by variations in the minor sex pheromone components and in the host plants, as well as by changes in the ratio of the two major components.     

Monitoring oriental fruit moth (Lepidoptera: Tortricidae) with the Ajar bait trap in orchards under mating disruption

Studies in Oregon, California, Pennsylvania and Italy evaluated the relative performance of the Ajar trap compared with several other traps for the capture of Grapholita molesta (Busck), in pome and stone fruit orchards treated with sex pheromone dispensers for mating disruption. The Ajar is a delta‐shaped trap with a screened jar filled with an aqueous terpinyl acetate plus brown sugar bait solution (TAS) that opens inside the trap and is surrounded by a sticky liner. The TAS‐baited Ajar trap was evaluated with and without the addition of a sex pheromone lure and compared with a delta trap baited with a sex pheromone lure and a bucket trap filled with the TAS bait. Although the Ajar trap had a 90% lower evaporation of the TAS bait than the bucket trap, both of them caught similar numbers in the majority of the field tests of both sexes of G. molesta. The addition of the sex pheromone lure did not increase moth catches by the TAS‐baited Ajar trap. The TAS‐baited Ajar trap caught significantly greater numbers of moths than the sex pheromone‐baited delta trap in 18 of the 20 orchards. Few hymenopterans were caught in orange TAS‐baited Ajar traps, but the catch of flies and other moths relative to the target pest remained high. Flight tunnel and field tests evaluated the effect of several screen designs on the catches of G. molesta and non‐target species. All exclusion devices significantly reduced the catch of larger moths. However, designs that did not reduce the catch of male G. molesta did not reduce the catch of muscid flies. Exclusion devices with openings <7.0 mm significantly reduced the catch of female G. molesta. The addition of (E)‐β‐farnesene, (E)‐β‐ocimene or butyl hexanoate septa lures to TAS‐baited Ajar traps significantly increased total moth catch. The addition of (E)‐β‐ocimene also significantly increased female moth catch.     

Monitoring oriental fruit moth and codling moth (Lepidoptera: Tortricidae) with combinations of pheromones and kairomones

Experiments were conducted in North and South America during 2012–2013 to evaluate the use of lure combinations of sex pheromones (PH), host plant volatiles (HPVs) and food baits in traps to capture the oriental fruit moth, Grapholita molesta (Busck), and codling moth, Cydia pomonella (L.), in pome and stone fruit orchards treated with sex pheromones. The combination of the sex pheromone of both species (PH combo lure) significantly increased G. molesta and marginally decreased C. pomonella captures as compared with captures of each species with either of their sex pheromones alone. The addition of a HPV combination lure [(E,Z)‐2,4‐ethyl decadienoate plus (E)‐β‐ocimene] or acetic acid used alone or together did not significantly increase the catch of either species in traps with the PH combo lure. The Ajar trap baited with terpinyl acetate and brown sugar (TAS bait) caught significantly more G. molesta than the delta trap baited with PH combo plus acetic acid in California during 2012. The addition of a PH combo lure to an Ajar trap significantly increased catches of G. molesta compared to the use of the TAS bait or PH combo lure alone in 2013. Female G. molesta were caught in TAS‐baited Ajar traps at similar levels with or without the use of additional lures. Ajar traps baited with the TAS bait alone or with (E)‐β‐ocimene and/or PH combo lures caught significantly fewer C. pomonella than delta traps with sex pheromone alone. Ajar traps with 6.4‐mm screened flaps caught similar numbers of total and female G. molesta as similarly baited open Ajar traps, and with a significant reduction in the catch of non‐targets. Broader testing of HPV and PH combo lures for G. molesta in either delta or screened or open Ajar traps is warranted.     

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.     

Two terpenoids activates close mating behavior and enhances trap efficiency of sex pheromone of Grapholita molesta

Grapholita molesta (Busck) (Lepidoptera: Tortricidae) is a notorious pest of many Rosaceae crops worldwide. Enhancement of trap efficiency of its sex pheromone was devised by addition of E-β-ocimene and E-β-farnesene. The addition of E-β-ocimene or E-β-farnesene to sex pheromone increased electroantennogram response of male G. molesta compared to sex pheromone alone. Blend of pheromone and E-β-ocimene or E-β-farnesene in 1:0.1 increased the upwind flight and landing behaviors. Furthermore, field experiments showed that sex lures with E-β-ocimene, or /and E-β-farnesene, enhanced trapping efficiency compared to sex pheromone alone. These results may provide the basis for the development of efficient pest management systems against G. molesta using plant volatiles and insect sex pheromones.     

Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures

Male and female moth catches of Grapholita molesta (Busck) in traps were evaluated in stone and pome fruit orchards untreated or treated with sex pheromones for mating disruption in Uruguay, Argentina, Chile, USA, and Italy from 2015 to 2017. Trials evaluated various blends loaded into either membrane cup lures or septa. Membrane lures were loaded with terpinyl acetate (TA), acetic acid (AA) and (Z)‐3‐hexenyl acetate alone or in combinations. Two septa lures were loaded with either the three‐component sex pheromone blend for G. molesta alone or in combination with codlemone (2‐PH), the sex pheromone of Cydia pomonella (L). A third septum lure included the combination sex pheromone blend plus pear ester, (E,Z)‐2,4‐ethyl decadienoate (2‐PH/PE), and a fourth septum was loaded with only β‐ocimene. Results were consistent across geographical areas showing that the addition of β‐ocimene or (Z)‐3‐hexenyl acetate did not increase moth catches. The addition of pear ester to the sex pheromone lure marginally increased moth catches. The use of TA and AA together significantly increased moth catches compared with the use of only one of the two components. Traps with the TA/AA lure outperformed the Ajar trap baited with a liquid TA plus sugar bait. The emission rate of AA was not a significant factor affecting the performance of the TA/AA lure. The addition of TA/AA significantly increased moth catches when combined with the 2‐PH lure. The TA/AA lure also allowed traps to catch both sexes. Catch of C. pomonella with the 2‐PH lure was comparable to the use of codlemone; however, moth catch was significantly reduced with the 2‐PH/PE lure. Optimization of these complex lures can likely further improve managers’ ability to monitor G. molesta and help to develop multispecies tortricid lures for use in individual traps.     

Real-time monitoring of oriental fruit moth,Grapholita molesta,populations using a remote sensing pheromone trap in apple orchards

A fusion of information technology (IT) and sex pheromone monitoring provides a remote sensing IT-pheromone trap to monitor Oriental fruit moth, Grapholita molesta, populations in apple orchards. Once a male of G. molesta is attracted to its sex pheromone lure in the trap, an infrared sensor installed at the funnel-shaped orifice generates an electric signal. The signal is processed in a central processor and then transferred to an internet site via a code division multiple access protocol. The signal also contains information about when each male is caught. Daily trapping information from different localities is archived in a website. The accuracy of IT-pheromone traps in detecting male catches was shown by a high correlation (r = 0.956) between the generated IT signals and actual numbers of males caught in the trap in apple orchards. Using this IT-pheromone trap, G. molesta in apple orchards was monitored for one year. These data were compared with monitoring data obtained from a conventional wing type-based sticky trap containing the identical sex pheromone lure. Both showed four characteristic adult peaks from April to September and were significantly correlated (r = 0.695). IT-pheromone traps also gave real-time signals of male catches in the field. These real-time signals of male catches showed a characteristic diel attraction rhythm from 4 pm to midnight. The diel rhythm of the male response to the sex pheromone started earlier in the evening in the spring season compared to mid and late seasons. This study provides a novel sex pheromone trap for G. molesta to monitor its population in field conditions in real-time without visiting or counting. The field monitoring data can be accessed any time through a designated internet website.     

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.     

Sexual differences in the flight performance of the oriental fruit moth, Cydia molesta

Cydia molesta Busck (Lepidoptera: Tortricidae), a major pest of stone fruits and an increasingly important late season pest of apple, is predominately monitored by pheromone trapping of male moths. We investigated flight performance in relation to sex, age, and mating status using computer-linked flight mills, and also examined the relationship between female flight and reproduction. The crepuscular flight pattern of the experimental moths in relation to photointensity was very similar to that reported from field studies. Female moths significantly outperformed males in all measured flight parameters including total distance flown, distance of longest single flight, and velocity. The proportion of long-flying females (categorised as those completing an unbroken flight of greater than 1 km) was three to six times greater than that of males. Female flight performance was not related to mating status, but mated males displayed significantly greater flight than unmated males. The maximal flight period of mated females commenced on the third day after eclosion, following 30% egg deposition. Male flight was not significantly related to age. The data suggest that a limited proportion of the population, in particular females, may have the capacity to make inter-orchard flights. The limitations of monitoring C. molesta populations in apple orchards solely by pheromone trapping of males are discussed.     

Behaviour of male oriental fruit moth, Grapholita molesta, in overlapping sex pheromone plumes in a wind tunnel

The behaviour of Grapholita molesta (Busck) (Lepidoptera: Tortricidae) males was studied in overlapping sex pheromone plumes in a wind tunnel. The ultimate aim of the study was to assess the effect of different treatments on male behaviour and consider the observed changes within the context of the suggested mechanisms underlying mating disruption. Two baits were placed either in series or parallel using both synthetic pheromone blends and female extracts. One bait, the reference containing (Z)-8-dodecenyl acetate/(E)-8-dodecenyl acetate/(Z)-8-dodecenol in a ratio of 100/6/10 was kept constant at a dose of 100 g of the main component, giving a composition and a release rate close to that of a female. The dose of the other bait varied between 0.1 and 100 times the concentration of the reference and was a mixture of one, two or three pheromone components. Males clearly discriminated between different blends and doses in the overlapping plumes, for regardless if the lures were presented in series or in parallel they followed the complete plume. Complete suppression of the response to the reference was only achieved with 300 g of the optimal three-component blend on the other lure. When tested singly, a bait consisting of Z8-12:OAc/E8-12:OAc/Z8-12:O Hin a 100/0.2/0.4 ratio, attracted a high proportion of the males when placed 75 cm upwind of the male release site, but no males from 150 cm. Our data suggest that complete pheromone blends should be more effective than any incomplete blends in mating disruption formulations for G. molesta.     

Trapping and sex pheromone-mediated flight and landing behaviour of maleCtenopseustis obliquana

A sex pheromone-baited delta trap was found to be inefficient at eliciting landing and entering of the trap by maleCtenopseustis obliquana. The inefficiency of the delta trap related to turbulence altering the pheromone plume and the concomitant effect on the flight manoeuvres of male moths. In the wind tunnel, high proportions of males flew upwind and landed on the sides, outside, of the trap, but only a relatively small proportion of these males entered the trap and contacted the sticky surface. When males approached the delta trap, they tended to fly in wide zigzags (i.e., large inter-track reversal distances) and at an altitude near the top of the trap, where the trap was relatively narrow in width (compared to the bottom). Thus, these flight manoeuvres largely precluded males from entering the trap. Greater numbers of male moths entered the trap when: (i) the front barriers of the delta trap were removed, (ii) pheromone dosage was increased to 300 μg, and (iii) the trap design was changed to a rectangular one. The first two changes appeared to influence the flight manoeuvres of males (who appeared to fly with narrower inter-track reversal distances), while the third change apparently did not affect the glight manoeuvres of males, but rather allowed more males to enter the trap because of the greater area of the entrance. The low trap catches of maleC. obliquana in the field are also a consequence of the glue which is currently used. After contact with this glue most males are able to escape, flying off the sticky surface and losing contact with the pheromone. A field trial found that delta traps with another glue caught more than three times the number of males of the related tortricid moth,Epiphyas postvittana, than delta traps with the currently used glue.     

A sex pheromone component novel to the Geometridae identified fromEpirrita autumnata

(Z,Z,Z)-1,3,6,9-Heneicosatetraene was identified as a sex pheromone component ofEpirrita autumnata (Borkhausen), (Lepidoptera: Geometridae). Gas chromatography coupled with electroantennographic detection revealed one active peak from female pheromone gland extracts. The chemical identification of the compound was based on gas chromatography-mass spectrometry, chemical micro-reactions and unambiguous synthesis. In a field test, a rubber septum loaded with 3000 μg of this compound showed highest attraction of maleE. autumnata moths compared to lower doses and the control. The attraction of males to a 300 μg bait was equivalent to that obtained using live virgin females.     

THE EVOLUTION OF SOCIAL SIGNALS: MORPHOLOGICAL,FUNCTIONAL, AND GENETIC INTEGRATION OF THE SEX PHEROMONE IN NAUPHOETA CINEREA

Social signals that mediate intraspecific interactions can be complex, conveying considerable information concerning the probable behavior of individuals and minimizing overt aggression and wasted energy. In the cockroach Nauphoeta cinerea, male-male competition and female mate choice are mediated by a multicomponent male-produced sex pheromone. In this study, I examine variation in this pheromone. First I measure differences among males in both individual pheromone compounds and the overall composition of the pheromone. Principal component analysis is used to quantify and describe pheromone composition. Next, I explore some of the causes and consequences of this variation by examining the pheromone of males with different social experiences. Compared to subordinate males, dominant males have significantly less variable quantities of the individual pheromone compounds and are significantly less variable in the composition of their pheromone. Because of an association between status and mating success, male-male competition can result in stabilizing sexual selection on the sex pheromone. Finally, I test the hypothesis that the pheromone compounds evolve in a manner consistent with their function. As predicted for morphologically integrated characters, the patterns of phenotypic, genetic, and environmental correlations among my measures of pheromone compounds and composition match functional patterns suggested by this study and the developmental patterns demonstrated in my previous studies. Based on these studies of the N. cinerea sex pheromone, I argue that stabilizing sexual selection shapes the evolution of pheromonal communication involved in social interactions among male N. cinerea. Further, I argue that coordinated evolution of social signals may be possible due to the morphological integration of their multiple compounds.     

Rapid intracellular alkalinization of Saccharomyces cerevisiae MATa cells in response to α-factor requires the CDC25 gene product

The α-factor mating pheromone induces a transient intracellular alkalinizatin of MATa cells within minutes after exposure to the pheromone, and is the earliest biochemical event that can be identified subsequent to the exposure. Dissipation of the pheromone induced pH gradient, using 2,4-dinotrophenol or sodium orthovanadate, does not inhibit the biological response of the yeast to the pheromone such as mating and ‘schmoo’ formation. These findings suggest that the pheromone mediated pH change per se is not a part of the transmembrane signalling but rather the consequence of a biochemical reaction triggered by the α-pheromone interaction with its receptor and may have a permissive effect on the pheromonal response. The cdc25^ts mutation causes MATa cells to become nonresponsive to α-factor subsequent to a shift to the restrictive temperature, suggesting that the CDC25 gene product participates in the pheromone response pathway.     

Field observations quantifying attraction of four tortricid moths to high-dosage pheromone dispensers in untreated and pheromone-treated orchards

Orientational responses of four species of feral tortricid moths (Lepidoptera: Tortricidae) to polyethylene tube dispensers of pheromone were observed in a 0.8 ha apple orchard treated with such pheromone dispensers and in an untreated 0.8 ha orchard. Male oblique‐banded leafrollers, Choristoneura rosaceana (Walker) (mean 7.2 ± 0.4 moths/night during 21 nights), Oriental fruit moths, Grapholita molesta (Busck) (mean 10.5 ± 2.1 during 20 evenings), and the redbanded leafrollers, Argyrotaenia velutinana (Walker) (mean 2.0 ± 1.1 during 14 nights) were observed approaching within 100 cm of their respective polyethylene‐tube pheromone dispensers in the untreated orchard. Furthermore, C. rosaceana (mean 2.0 ± 0.7 during 17 nights) and G. molesta (mean 1.5 ± 0.4 over 20 evenings) came within 100 cm of their respective polyethylene‐tube pheromone dispensers in the pheromone‐treated orchard. Most visits lasted less than 10 s, after which the majority of moths departed by flying upwind. In the untreated orchard, the number of C. rosaceana observed orienting to polyethylene tube dispensers was greater than the number captured in optimized monitoring traps (1.9 ± 0.4) per night of observation. The numbers of A. velutinana (2.0 ± 1.1) or G. molesta (10.5 ± 2.1) attracted to polyethylene‐tube dispensers in the untreated orchard did not differ statistically from the numbers captured in optimized monitoring traps per night of observation. In the pheromone‐treated orchard, the number of C. rosaceana (2.0 ± 0.4) or G. molesta (1.2 ± 0.2) observed orienting to polyethylene‐tube dispensers did not differ statistically from the numbers of male moths of these species captured in optimized monitoring traps per night of observation. No codling moths, Cydia pomonella L. were observed orienting to, or landing near, their respective polyethylene‐tube dispensers in either the untreated or pheromone‐treated orchards, although substantial numbers were captured in monitoring traps per night of observation (6.0 ± 1.7) in the untreated orchard. The attraction of male moths to polyethylene tube dispensers thus occurred in three of the four species observed. These results provide support for the idea that false‐plume‐following is an important component of the mechanisms mediating communicational disruption in moths by polyethylene‐tube dispensers.     

Representation of a mixture of pheromone and host plant odor by antennal lobe projection neurons of the silkmoth Bombyx mori

Pheromone-source orientation behavior can be modified by coexisting plant volatiles. Some host plant volatiles enhance the pheromonal responses of olfactory receptor neurons and increase the sensitivity of orientation behavior in the Lepidoptera species. Although many electrophysiological studies have focused on the pheromonal response of olfactory interneurons, the response to the mixture of pheromone and plant odor is not yet known. Using the silkmoth, Bombyx mori, we investigated the physiology of interneurons in the antennal lobe (AL), the primary olfactory center in the insect brain, in response to a mixture of the primary pheromone component bombykol and cis-3-hexen-1-ol, a mulberry leaf volatile. Application of the mixture enhanced the pheromonal responses of projection neurons innervating the macroglomerular complex in the AL. In contrast, the mixture of pheromone and cis-3-hexen-1-ol had little influence on the responses of projection neurons innervating the ordinary glomeruli whereas other plant odors dynamically modified the response. Together this suggests moths can process plant odor information under conditions of simultaneous exposure to sex pheromone.     

Enzymatic Reduction of Sorbic Acid to trans-4-Hexmol

The sex pheromone of the citrus mealybug (Pseudococcus cryptus), [(1R,3R)-3-isopropenyl-2,2-dimethylcyclobutyl]methyl 3-methyl-3-butenoate, was synthesized from (+)-α-pinene in five operational steps in a 43% overall yield. The synthetic pheromone was identical with the natural pheromone in ¹H-NMR and mass spectroscopic properties, and showed almost the same pheromonal activity as the natural pheromone.     

Male Aggression,Female Mimicry and Female Choice in the Rove Beetle,Aleochara curtula (Coleoptera,Staphylinidae)

Causes, consequences and pheromonal regulation of male contest and female choice in the staphylinid beetle, Aleochara curtula (Goeze), have been investigated in the field and in the laboratory. At the feeding and mating site (carcass) the sex ratio is male biased. Polyandry is affected by prolonged copulations, spermatophore plugs and anti-aphrodisiac pheromones transferred from the male, and by female repulse behaviour as well. Aggression of competing males leads to expulsion of inferior males from the carcass. Young, starved and multiply mated males, which need access to the food resource, produce the female sex pheromone. They release homosexual responses, but also avoid intrasexual aggression. On the other hand, females behave aggressively towards individuals bearing the female sex pheromone or repulse their copulatory attempts. Those males of insufficient physiological condition produce a lighter spermatophore and fertilize less eggs. The adaptive significance of female mimicry, male mating tactics, and female choice is discussed.     

Yeast engineering to express sex pheromone gland genes of the oriental fruit moth,Grapholita molesta

Mating disruption by using sex pheromone is an ecofriendly alternative way to control insect pests. To be effective, large amounts of sex pheromone are needed, leading to a relatively high production cost. To reduce the cost for chemical synthesis of sex pheromone, yeast engineering technology has been devised. This study used a baker's yeast, Saccharomyces cerevisiae, to express genes associated with sex pheromone biosynthesis of the Oriental fruit moth, Grapholita molesta. Compared to other fatty acid biosynthetic pathways, two steps that are unique to pheromone gland of G. molesta are proposed: desaturation at even number catalyzed by desaturase (Gm-DES) and terminal reduction catalyzed by fatty acyl reductase (Gm-FAR). Gm-DES and Gm-FAR were cloned into a yeast expression vector, pYES2.1. They were used to transform S. cerevisiae by a double transfection method. The transformed yeast was induced with 2% galactose to over-express these two exogenous genes. Their expression was confirmed by RT-PCR and western blotting. To facilitate pheromone production, transformed yeasts were supplied with myristic acid during over-expression. Resulting fatty acid composition was analyzed by GC-MS after fatty acid methyl ester derivatization. Control yeast produced mostly saturated fatty acids. However, a single gene (Gm-DES)-transformed yeast produced unsaturated fatty acids at ∆9 such as Z9-tetradecenoic acid (Z9-14:1), palmitoleic acid (Z9-16:1), and oleic acid (Z9-18:1) in addition to saturated fatty acids. The double-transformed yeast produced an additional component, alcohol form of oleic acid (Z9-18:OH). These results suggest that Gm-DES can catalyze desaturation of fatty acids at ∆9 and Gm-FAR can reduce terminal carboxylic acid into alcohol.