Pheromone pre-exposure and mating modulate codling moth (Lepidoptera: Tortricidae) response to host plant volatiles

Abstract 1 Two codling moth Cydia pomonella kairomonal attractants, ethyl (E,Z)‐2,4‐decadienoate (pear ester) and (E)‐β‐farnesene, were tested in an insecticide‐sprayed apple orchard and an orchard treated for mating disruption with synthetic pheromone (E,E)‐8,10‐dodecadienol (codlemone). Male captures with pear ester were higher in the pheromone‐treated than in the insecticide‐treated orchard, whereas captures with (E)‐β‐farnesene were not different. Subsequent wind tunnel experiments confirmed that pre‐exposure to sex pheromone codlemone increased the behavioural response of codling moth males to pear ester. This supports the idea that male attraction to the plant volatile pear ester and sex pheromone codlemone is mediated through the same sensory channels. 2 Pear ester is a bisexual codling moth attractant and even captures of female moths were significantly increased in the pheromone‐treated orchard. In the laboratory wind tunnel, pheromone pre‐exposure had no effect on female response to pear ester, but significantly more mated than unmated codling moth females flew upwind towards a pear ester source. Differences in mating status in insecticide‐treated vs. pheromone‐treated orchards may thus account for the differences in female trap captures with pear ester. 3 These findings are important with respect to monitoring of codling moth with pear ester in mating disruption orchards. They also emphasize the importance of host plant volatiles in pheromone‐mediated mating disruption, which has been neglected to date.     

Monitoring codling moth (Lepidoptera: Tortricidae) in sex pheromone-treated orchards with (E)-4,8-dimethyl-1,3,7-nonatriene or pear ester in combination with codlemone and acetic acid

Traps baited with ethyl (E,Z)-2,4-decadienoate (pear ester) or (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) in two- or three-way combinations with the sex pheromone (E,E)-8,10-dodecadien-1-ol (codlemone) and acetic acid (AA) were evaluated for codling moth, Cydia pomonella (L.). All studies were conducted in apple orchards, Malus domestica Borkhausen, treated with sex pheromone dispensers during 2010. Septa were loaded with codlemone, DMNT, and pear ester individually or codlemone with either DMNT or pear ester together (combo lures). Polyethylene vials loaded with AA were added as a co-lure. Residual analyses of field-aged combo lures and weight loss of the AA co-lure were conducted. AA vials lost 50-150 mg wk(-1). Weekly weight loss was not affected by field aging, but was closely correlated with the daily mean temperature. Pear ester was released from septa at a slightly higher but nonsignificant rate than codlemone. DMNT was released at a significantly higher rate than codlemone, and lures were effective for 4 wk. The addition of codlemone to traps baited with either host plant volatile plus AA had either no effect or significantly increased total moth catches. The addition of AA significantly increased the catch of female moths with either combo lure. Total moth catches in traps baited with pear ester or DMNT combo lures and AA did not differ and were either significantly higher or similar to the pear ester combo lure. These data suggest that codling moth may be more effectively monitored in sex pheromone-treated orchards with multi-component lures, including codlemone, AA, and host plant volatiles.     

Antennal responses of Cydia pomonella (L.) exposed to surfaces treated with methoxyfenozide

Electroantennogram (EAG) measurements were recorded from the antennae of male and female codling moth, Cydia pomonella L., to determine whether adult moths exposed to surfaces treated with the ecdysteroid agonist methoxyfenozide experience a decline in their antennal reception and thus olfactory sensitivity. Such a phenomenon would offer a possible mechanism for the previously reported decreased responsiveness from moths treated with methoxyfenozide to pheromone‐ and plant volatile‐based monitoring lures. Mean EAG data revealed that the antennae from methoxyfenozide‐treated male moths appear to be just as sensitive to various doses of synthetic codlemone as the antennae from the control and surfactant‐treated moths, but they appeared to be less sensitive to the pheromone component 12OH (collected from female effluvia) than the control male antennae. Mean male EAG responses to the pheromone components E8,E10‐12Al and codlemone collected from methoxyfenozide‐treated females were significantly less than the responses towards those two pheromone components collected from the control and surfactant‐treated females. Female moth exposure to methoxyfenozide did not negatively impact the sensitivity of female antennae to the plant volatile pear ester, but it did towards the apple volatile butyl hexanoate. Data from this study show that adult C. pomonella exposure to methoxyfenozide‐treated surfaces does appear to negatively impact, in a minor way, the (i) olfactory sensitivity (or detection) of male antennae towards some components of the female sex pheromone, (ii) the female antennal sensitivity towards a key apple volatile and (iii) the attractiveness of female pheromone effluvia.     

Flight tunnel response of codling moth Cydia pomonella to blends of codlemone,codlemone antagonists and pear ester

Upwind orientation flights of codling moth males Cydia pomonella L. to a single source of sex pheromone (E,E)‐8,10‐dodecadienol (codlemone) are significantly reduced when blending it with pheromone antagonists, either with codlemone acetate, (E,E)‐8,10‐dodecadienyl acetate, or with the codlemone isomer (E,Z)‐8,10‐dodecadienol. However, once activated by a pheromone stimulus, males no longer distinguish between a pheromone source and these antagonistic blend sources. This shows that the pheromone stimulus required for the initiation of an upwind flight response differs from the stimulus for maintaining upwind flight and landing at the source. In contrast to pheromone antagonists, males discriminate between pheromone alone and a blend source of pheromone and the plant volatile pear ester, ethyl (2E,4Z)‐2,4‐decadienoate. This indicates a difference in the detection and neural integration of pheromone and plant volatile stimuli.     

Exposure to methoxyfenozide-treated surfaces reduces the responsiveness of adult male codling moth (Lepidoptera: Tortricidae) to codlemone and pear ester lures in a wind tunnel

The responsiveness of male codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), exposed to surfaces treated with the ecdysteroid agonist methoxyfenozide, toward lures loaded with the synthetic sex pheromone codlemone and/or the pear ester kairomone were investigated in wind tunnel experiments. Five different kinds of commercially available codling moth monitoring lures (obtained from Tr6c6 Inc., Adair, OK) were used in the bioassay: Pherocon CM Standard lure (loaded with 1 mg of codlemone), Pherocon CM Long-Life L2 (loaded with 3.5 mg of codlemone), Pherocon CM 10X (loaded with 10 mg of codlemone), Pherocon CM-DA Combo (loaded with 3.0 mg of codlemone and 3.0 mg of pear ester), and Pherocon DA (loaded with 3.0 mg of pear ester). Results from the study revealed that male codling moth exposed to surfaces treated with methoxyfenozide and the surfactant exhibited a significant decline in responsiveness toward lures loaded with either codlemone or pear ester. The full impact of how this negative effect might alter current moth monitoring procedures in orchards receiving ecdysone agonist sprays requires further investigation.     

Traps baited with pear ester and acetic acid attract both sexes of Hedya nubiferana (Lepidoptera: Tortricidae)

This study was initiated with the objective of studying field responses of the green budworm moth, Hedya nubiferana (Haworth) (Lepidoptera: Tortricidae), to pear ester [PE; ethyl (E,Z)‐2,4‐decadienoate] and acetic acid (AA) with the aim of developing a lure attractive also for females. In the overwhelming majority of tests, traps baited with the PEAA lure (the combination of both PE and AA) caught more than traps baited with either of the constituents presented alone. PEAA lures were attractive to H. nubiferana no matter whether the two compounds were provided in separate dispensers or mixed together in a single one, and a large percentage (up to 71%) of trap catch consisted of females. Traps with PEAA lures caught (females plus males) on an average 30% of the catches in traps baited with the synthetic green budworm moth sex pheromone (only males). This suggested that the new PEAA lure had a trapping performance comparable with that of pheromone traps, which latter are used by farmers today. Consequently, the PEAA lure showed potential for future practical applications as a female‐targeted lure for H. nubiferana. To our knowledge, this is the first well‐documented report on the attraction of PEAA lure for a tortricid species other than codling moth.     

Odour quality discrimination for behavioural antagonist compounds in three tortricid species

The antennal and behavioural response of three tortricid species (Lepidoptera: Tortricidae) to their corresponding sex pheromones and known or putative behavioural antagonists was tested by electroantennography and in field trials. The species and their pheromones and known or proposed behavioural antagonist were lightbrown apple moth, Epiphyas postvittana (Walker) [pheromone: 95% (E)‐11‐tetradecenyl acetate (E11‐14Ac) and 5% (E,E)‐9,11‐tetradecadienyl acetate (E9E11‐14Ac); antagonist: (Z)‐11‐tetradecenyl acetate (Z11‐14Ac)], codling moth, Cydia pomonella (L.) [pheromone: (E,E)‐8,10‐dodecadien‐1‐ol (codlemone); antagonist: (E,E)‐8,10‐dodecadienyl acetate (codlemone acetate)], and gorse pod moth, Cydia ulicetana (Haworth) [pheromone: (E,E)‐8,10‐dodecadienyl acetate (codlemone acetate); putative antagonist: (E,E)‐8,10‐dodecadien‐1‐ol (codlemone)]. In all three species, the antennal response to the antagonists was not significantly different from the antennal response to con‐specific sex pheromone compounds. In the field trapping experiments, significantly fewer males of all three species were attracted to the respective pheromone when blended with the behavioural antagonist compound. However, this response varied between the species, with lightbrown apple moth and codling moth showing stronger responses to the antagonist compounds than gorse pod moth. Both lightbrown apple moth and codling moth males were able to discriminate between pure pheromone and pheromone blended with the antagonist when placed in traps side‐by‐side separated by ca. 10 cm. The presence of the behavioural antagonist not only affected the catch of males of both species within their own traps but also affected the catch in the neighbouring trap that contained con‐specific sex pheromone; the catch of gorse pod moth was not reduced by the presence of codlemone in the neighbouring trap. These results suggest that strong behavioural antagonists such as codlemone acetate for codling moth and Z11‐14Ac for lightbrown apple moth induce their inhibition effect at a substantial distance downwind from the odour source; however, most of those males that were able to overcome this inhibition effect at the early stage of orientation to odour source, were able to discriminate between the pheromone source and the pheromone source admixed with behavioural antagonist. Moderate behavioural antagonists such as codlemone for gorse pod moth did not elicit a discrimination effect.     

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

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

Electroantennogram responses of Plutella xylostella (L.), to sex pheromone components and host plant volatile semiochemicals

The diamondback moth (DBM), Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a notorious insect pest of cruciferous crops worldwide. Attract-and-kill strategies to manage the DBM based on insect pheromone and plant volatile semiochemicals have been explored and partially applied in the field. However, little is known about whether the attractant effects of insect pheromone and host plant volatile semiochemicals on insects are affected by insect age or volatile concentrations. Therefore, we examined the electroantennogram (EAG) responses of both DBM males and females varying in age and body size to a range of concentrations of seven host plant volatiles and two main female sex pheromone components. Our results showed that DBM age had no influence on EAG responses to trans-2-Hexenal, trans-2-Hexenol, Heptanal, cis-3-Hexen-1-ol and Z11-16:Ald and significantly influenced the EAG responses to Z11-16:Ac and three isothiocyanates. Age and sex had interaction effects on EAG responses of DBMs to some tested semiochemicals. DBM females had significantly stronger responses than males to all tested plant volatiles, and values were enhanced with increasing concentrations. Moreover, the degree of enhancement of EAG responses in females was larger than that in males for trans-2-Hexenal, trans-2-Hexenol, cis-3-Hexen-1-ol, 2-Phenylethyl isothiocyanate and Methyl isothiocyanate treatments. Body size seemed to have no influence on EAG responses. Our results might provide a theoretical basis for optimizing attract-and-kill strategies for insect pests.     

农业昆虫气味受体功能研究进展

昆虫主要依靠嗅觉系统寻找食物、发现配偶、控制交配、选择产卵地、逃避天敌等,因此嗅觉系统对昆虫的繁殖和生存至关重要。气味受体(odorant receptor, OR)是嗅觉系统中的关键成分之一,可被信息化合物激活引发特定行为产生。随着测序技术的发展,大量的农业昆虫的基因组和转录组被测序,从测序数据中分析获得了它们的OR家族。现在OR功能研究经常用到外源表达系统和CRISPR/Cas9系统。外源表达系统结合记录系统可用于目标OR表达和配体筛选,CRISPR/Cas9系统可从昆虫染色体上敲除目标OR基因,进而利用电生理技术和行为学实验研究其功能。本文系统总结了鳞翅目、直翅目、半翅目、双翅目、膜翅目和鞘翅目6个目30个种的农业昆虫OR的气味分子反应谱及功能,尤其是鳞翅目农业昆虫。农业昆虫的性信息素往往是由雌虫产生的两种或两种以上成分按一定比例组成的混合物,包括在种间生殖隔离中起作用的行为拮抗剂。每个种因此具有多个性信息素的受体,共同感受这些信息素信息,调控种内和种间性行为。有些OR可调谐至主要针对植物挥发物,包括花香气味,在寄主植物选择和雌虫选择产卵地等行为中起作用。聚集信息素受体可特异地被聚集信息素激活,引起聚集行为;报警信息素受体可特异地被报警信息素激活,引起排斥行为。研究农业昆虫OR的气味分子反应谱和功能可为开发在害虫防控中应用的性诱剂、食诱剂、拒食剂、聚集信息素等奠定理论基础。最后我们对未来的主要研究方向提出了以下建议：(1)开发新的外源OR表达系统;(2)研究在雌虫中特异调谐至针对雄虫产生的性信息素的性信息素受体的功能;(3)探讨OR与气味分子之间特异性相互作用的分子机制。     

Evaluating dispensers loaded with codlemone and pear ester for disruption of codling moth (Lepidoptera: Tortricidae)

Polyvinyl chloride polymer (PVC) dispensers loaded with ethyl (E,Z)-2,4-decadienoate (pear ester) plus the sex pheromone, (E,E)-8,10-dodecadien-1-ol (codlemone) of codling moth, Cydia pomonella (L.), were compared with PVC dispensers and a commercial dispenser (Isomate-C Plus) loaded with codlemone. Evaluations were conducted in replicated plots (0.1-0.2 ha) in apple, Malus domestica (Borkhausen) during both generations of codling moth from 2007 to 2009. Dispensers were applied at 1,000 ha(-1). Male captures in traps baited with virgin female moths and codlemone lures were recorded. Residual analysis of field-aged dispensers over both moth generations was conducted. Dispensers exhibited linear declines in release rates of both attractants, and pear ester was released at a significantly higher rate than codlemone during both time periods. The proportion of virgin female-baited traps catching males was significantly lower with combo dispenser TRE24 (45/110, mg codlemone/mg pear ester) during the second generation in 2007 and the combo dispensers TRE144 (45/75) and TRE145 (75/45) during the first generation in 2008 compared with Isomate-C Plus. Similarly, male catches in female-baited traps in plots treated with the combo dispensers TRE144 during the first generation in 2008 and TRE23 (75/110) during the second generation, in 2007 were significantly lower than in plots treated with Isomate-C Plus. No significant differences were found for male catches in codlemone-baited traps in plots treated with Isomate-C Plus and any of the combo dispensers. However, male catches were significantly lower in plots treated with Cidetrak CM (codlemone-only dispenser) than the combo TRE144 dispenser during both generations in 2009.     

植物源挥发物对昆虫信息素的增效作用及其增效机制

植物源挥发物和昆虫信息素是昆虫的重要信息物质,二者协同作用以调节昆虫的行为.通过增加触角电位、信息素接收神经元动作电位和脉冲频率,特异性植物源挥发物能显著增强昆虫性信息素和聚集信息素的引诱力.这种对昆虫信息素的增效作用受昆虫体内的章鱼胺及其受体介导.特异性植物源挥发物和章鱼胺受体结合,降低性信息素接收神经元对性信息素的反应阈值,增强性信息素接收神经元敏感性.这可能是植物源挥发物对昆虫信息素具有增效作用的主要机制.     

Combined approaches using sex pheromone and pear ester for behavioural disruption of codling moth (Lepidoptera: Tortricidae)

Attractive properties of pear ester, ethyl (E,Z)‐2,4‐decadienoate, and codlemone, (E,E)‐8,10‐dodecadien‐1‐ol, the sex pheromone of codling moth, Cydia pomonella (L.), were utilized in experiments on behavioural disruption of mating. Standard dispensers loaded with codlemone alone or in combination with pear ester (combo) were applied at 500–1000/ha. Larger (10‐fold) combo dispensers (Meso) were evaluated at a rate of 80/ha. The addition of microencapsulated pear ester, PE‐MEC, sprayed with insecticides at 30 ml/ha was also evaluated. Male moth catches in unmated female‐baited traps were lower in standard combo dispenser than in codlemone dispenser–treated plots. Female moth catch in traps baited with the combination of pear ester, codlemone and acetic acid was lower in standard combo dispenser than in codlemone dispenser–treated plots. In 12 comparative experiments spanning from 2006 to 2012, male moth catch in unmated female‐baited traps was consistently and significantly lower in combo than in codlemone dispenser–treated plots. Male catch in codlemone‐baited traps did not differ between dispenser treatments in eight studies from 2006 to 2009. These results emphasize the benefit of alternatively using traps baited with unmated females over codlemone lures for the analysis of dispenser activity. Fruit injury was significantly reduced with the addition of PE‐MEC to insecticide applications across untreated and dispenser treatments. Proportion of unmated females trapped was higher in standard combo dispenser than in codlemone dispenser–treated and untreated plots. Similarly, the proportion of unmated females caught was higher in the Meso combo dispenser than in nearby or distant codlemone dispenser–treated plots. These field studies conducted in apple over 3 years demonstrate that adding pear ester both to pheromone dispensers, either standard or Meso, and to supplementary insecticide sprays can provide a significant increase in the disruption of sexual communication, reductions in female mating and reductions in fruit injury.     

Behavioral observations of codling moth, Cydia pomonella, in orchards permeated with synthetic pheromone

Mating disruption of codling moth, Cydia pomonella, was studied in apple orchards treated with the main pheromone compound codlemone, (E,E)-8,10-dodecadienol, and a blend of codlemone and codlemone acetate, (E,E)-8,10-dodecadienyl acetate, a strong pheromone antagonist. Codlemone alone and the pheromone/antagonist-blend had a similar effect on the behavior of males emerging into air-permeated orchards. Male flights within tree canopy and upwind orientation along tree rows were strongly enhanced by both formulations, compared to untreated plots. However, the codlemone/codlemone acetate-blend increased the rate of cross-wind and downwind flights within the orchard, compared to codlemone alone. The major difference between these two formulations was that males from nearby, untreated orchards were attracted towards orchards treated with codlemone, but not towards treatments with codlemone/codlemone acetate. Additional tests were done with an equilibrium blend of codlemone and its geometric isomers. Aerial pheromone concentrations in the orchards were recorded by the field electroantennogram technique. Decreasing pheromone concentrations towards the upper part of the tree canopy, together with the stimulation of male flight activity by synthetic pheromone explains failures to control codling moth at high population densities with current dispenser formulations.     

Concurrent modulation of neuronal and behavioural olfactory responses to sex and host plant cues in a male moth

Mating has profound effects on animal physiology and behaviour, not only in females but also in males, which we show here for olfactory responses. In cotton leafworm moths, Spodoptera littoralis, odour-mediated attraction to sex pheromone and plant volatiles are modulated after mating, producing a behavioural response that matches the physiological condition of the male insect. Unmated males are attracted by upwind flight to sex pheromone released by calling females, as well as to volatiles of lilac flowers and green leaves of the host plant cotton, signalling adult food and mating sites, respectively. Mating temporarily abolishes male attraction to females and host plant odour, but does not diminish attraction to flowers. This behavioural modulation is correlated with a response modulation in the olfactory system, as shown by electro-physiological recordings from antennae and by functional imaging of the antennal lobe, using natural odours and synthetic compounds. An effect of mating on the olfactory responses to pheromone and cotton plant volatiles but not to lilac flowers indicates the presence of functionally independent neural circuits within the olfactory system. Our results indicate that these circuits interconnect and weigh perception of social and habitat odour signals to generate appropriate behavioural responses according to mating state.     

Sex pheromone of pear moth, Cydia pyrivora

The sex pheromone of the pear moth, Cydia pyrivora, is (E,E)-8,10-dodecadien-yl acetate. A 5%-addition of the geometric isomers E,Z and Z,Z decreased male attraction in the field, the Z,E isomer had no significant effect. Traps baited with 10 µg E,E on grey rubber septa were attractive throughout the flight period of C. pyrivora. These traps allow specific detection of pear moth, and they are not attractive to its sibling species, codling moth C. pomonella.     

信息化合物对昆虫行为的影响

本文综述了来自寄主植物的挥发性物质和同种昆虫或异种昆虫释放的各种信息素及两者的协同作用的信息化合物对昆虫行为的影响。特别强调了寄主植物的气味物质和昆虫信息素协同作用在昆虫寻找寄主、求偶、交配及天敌在寄主识别过程中的重要地位。昆虫对寄主植物的识别是由于识别了植物气味的由一定组分、按照严格比例组成的化学指纹图。昆虫信息素与植物挥发性物质相结合为昆虫寻找求偶、交配场所提供更复杂或更全面的信息。许多昆虫只有在寄主植物或寄主植物气味存在时 ,才能释放性或聚集信息素。天敌在寄主识别、搜索及定位等一系列过程中 ,来自寄主的食料、寄主本身及两者的互作的信息化合物起重要的作用。研究信息化合物对昆虫行为的影响可以探索昆虫各种行为的内在机理 ,更好的了解寄主—昆虫—天敌三层营养关系的相互作用 ,对利用天然活性化合物防治害虫及生物防治提供理论依据