Behavioural responses in three ichneumonid pollen beetle parasitoids to volatiles emitted from different phenological stages of oilseed rape

Pollen beetles (Meligethes spp.; Coleoptera: Nitiduliae) are a major pest of oilseed rape, Brassica napus L. (Brassicaceae) in northern Europe. Phradis interstitialis Thomson, P. morionellus Holmgr., and Tersilochus heterocerus Thomson (Hymenoptera: Ichneumonidae) are among the most frequent pollen beetle parasitoids. These three species differ in temporal occurrence, as well as in preferred host stage. The behavioural responses of female parasitoids to odours from oilseed rape at bud and flowering stage were evaluated in two‐choice experiments. The role of visual stimuli was examined by combining green and yellow colours with odour stimuli. All three species were attracted to odours from the bud stage of oilseed rape. Tersilochus heterocerus was attracted to odours of flowering rape, but the two Phradis species avoided the flower odours. However, when the odours of flowering rape were combined with yellow, and odours of the bud stage were combined with green, P. interstitialis was equally attracted to both stimuli, and T. heterocerus showed an increased preference for flower odours, while no effect of colours could be found in P. morionellus. The observed differences in responses between the parasitoids may reflect differences in their biology and may be involved in the niche segregation of these often coexisting species. The volatile blends released from the two phenological stages were identified and compared. Clearly, odours can be reliable cues for differentiating between oilseed rape in the bud and flowering stage. Of 20 identified compounds, 18 were released at a significantly higher rate from flowering plants. The terpenes sabinene, myrcene, limonene, and (E,E)‐α‐farnesene were the dominant volatiles in the bud and flower headspace. A group of aromatic compounds including benzaldehyde, methyl benzoate, and phenyl acetaldehyde were mainly released from flowering rape.     

Laboratory and field response of the pollen beetle, Meligethes aeneus, to the odour of oilseed rape

Abstract. The behavioural response of adults of the pollen beetle, Meligethes aeneus F. (Coleoptera: Nitidulidae), to the odour of one of their host-plants, oilseed rape ( Brassica napus L.), was observed in the laboratory using a fourarmed airflow olfactometer, and under semi-natural conditions using a markrelease–recapture technique. Oilseed rape leaf and flower odour was attractive to pollen beetles in the olfactometer and they dispersed upwind from the centre of a circular array (radius 20 m) of yellow water traps, baited with extracts of oilseed rape leaves or flowers. The dispersal of pollen beetles in the absence of oilseed rape odour was predominantly downwind and crosswind. The percentage recapture of pollen beetles was significantly greater in traps baited with rape flower extracts than leaf extracts. Pollen beetles can use odour-mediated upwind anemotaxis to locate oilseed rape plants from distances of at least 20 m over open ground.     

Divergent responses of two cereal aphids to previous infestation of their host plant

The role of pollen odour in resource location by the pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), a pollen-feeding insect regarded as a pest of oilseed rape, Brassica napus L., (Brassicaceae) crops, was investigated in a linear track olfactometer. Both male and female beetles were attracted to the odour of whole oilseed rape flowers, indicating that these insects can locate their host plants using floral odours as cues. The attractive odour of flowers was found to emanate from all floral parts tested: the petals/sepals, the anthers, and from pollen itself. Therefore, at least part of the attractive odour of oilseed rape flowers emanates from pollen. Beetles were more attracted to floral samples containing anthers than those without anthers when these odours were directly compared in a choice-test, and this indicates that there were detectable differences between them. Anthers and pollen may therefore release distinctive odours that are quantitatively and/or qualitatively different from the odour of the rest of the flower. These experiments support the hypothesis that pollen-seeking insects use pollen odour cues to locate this food source.     

Red oilseed rape? The potential for manipulation of petal colour in control strategies for the pollen beetle (Meligethes aeneus)

The pollen beetle (Meligethes aeneus) is a major pest of oilseed rape (Brassica napus) at the inflorescence stage and is well known to prefer colours called yellow by human observers over many other colours. While commercial cultivars of oilseed rape have yellow flowers, little is known about the potential to manipulate host plant location and reduce subsequent infestation by this pest through variation in flower colour. We investigated the responses of pollen beetles to flowers of a white-petalled oilseed rape variety that had been dyed different colours in semi-field arena and field experiments. Flowers dyed blue or red were less heavily infested than those dyed yellow or the white flowers, indicating that blue and red flowers were less attractive than yellow and white ones. This response was most likely due to differences in petal colour because olfactometer studies showed that beetle responses to the odours of the coloured treatments did not differ. The comparatively high infestation of untreated white flowers is interpreted as a consequence of their high UV reflectance; the presence of a UV receptor in M. aeneus is suggested, and its role in visually guided insect–plant interactions in this species described. The potential for manipulation of petal colour in control strategies for the pollen beetle is discussed.     

Exploitation of host plant preferences in pest management strategies for oilseed rape (Brassica napus)

New control strategies for insect pests of arable agriculture are needed to reduce current dependence on synthetic insecticides, the use of which is unsustainable. We investigated the potential of a simple control strategy to protect spring‐sown oilseed rape, Brassica napus L. (Brassicaceae), from two major inflorescence pests: the pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), and the seed weevil, Ceutorhynchus assimilis (Paykull) (Coleoptera: Curculionidae), through exploitation of their host plant preferences. The strategy comprised, for the main crop, Starlight [an oilseed rape cultivar with relatively low proportions of alkenyl glucosinolates in the leaves (thereby releasing lower levels of attractive isothiocyanates than conventional cultivars)] and turnip rape, Brassica rapa (L.) (Brassicaceae), as a trap crop. We tested the system in laboratory, polytunnel semifield arena, and field experiments. The odours of Starlight were less attractive in olfactometer tests to both pests than those from a conventional cultivar, Canyon, and the plants were less heavily colonized in both polytunnel and field experiments. Turnip rape showed good potential as a trap crop for oilseed rape pests, particularly the pollen beetle as its odour was more attractive to both pests than that of oilseed rape. Polytunnel and field experiments showed the importance of relative growth stage in the system. As turnip rape flowers earlier than oilseed rape, beetles would be maintained on turnip rape past the damage‐susceptible growth stage of oilseed rape. The development of a pest control regime based on this strategy is discussed.     

Impact of insecticides on oilseed rape bud infestation with eggs and larvae of pollen beetle (<Emphasis Type="Italic">Brassicogethes aeneus</Emphasis> (Fabricius))

One of the main insect pests in oilseed rape is the pollen beetle (Brassicogethes aeneus (Fabricius), syn. Meligethes aeneus). To maximize efficiency of control of this pest, insecticides are required that ideally, not just prevent yield losses by bud feeding of overwintered pollen beetles, but simultaneously minimize the reproduction of the pest, thereby reducing the size of the following generation infesting next year’s oilseed rape. The neonicotinoid active substance thiacloprid is known to reduce bud infestation with eggs and larvae. However, the mechanisms by which this occurs as well as the effects of other active substances are not known. In this study, the effects of the neonicotinoid insecticide Biscaya (a.i. thiacloprid) and the pyrethroids Mavrik (tau-fluvalinate) and Karate Zeon (lambda-cyhalothrin) applied at the bud stage of winter oilseed rape with recommended field rates on infestation of buds with eggs and larvae of pollen beetles were tested in field trials in Germany in 2013–2015. In additional greenhouse experiments, it was investigated whether effects on bud infestation were caused by lethal effects on pollen beetle or by insecticidal residues on plants causing sublethal effects. In the field trials, application of Biscaya and Mavrik significantly reduced the percentage of buds containing eggs and larvae in contrast to Karate Zeon. In 2014 and 2015, 14 days after application, bud infestation on the main raceme was reduced by 86 and 82%, respectively, in Biscaya-treated plots and by 51 and 71%, respectively, in Mavrik-treated plots compared to the untreated plots. In the greenhouse experiments, the lowest percentage of bud infestation with eggs and larvae was recorded on Biscaya-treated plants whereas on Mavrik-treated plants, there was no significant difference compared with the control. The results of the field trials show that Biscaya and Mavrik reduced oilseed rape bud infestation with eggs and larvae of pollen beetles primarily by lethal effects on overwintered pollen beetles or by repellency. However, Biscaya had additional effects on egg laying, which was supported by the greenhouse experiments.     

Glucosinolates in oilseed rape (Brassica napus) and the incidence of pollen beetles (Meligethes aeneus)

During an early stage of migration into a field of oilseed rape in the spring of 1988, more pollen beetles were observed on the double-low variety, Ariana, than on the adjacent single-low varieties, Bienvenu and Mikado. More beetles were found on open flowers on terminal racemes than on yellow and green flower buds on lateral racemes. Five alkenyl, three indolyl and two aromatic glucosinolates were identified in the floral tissues of oilseed rape. The major compounds were progoitrin (2-OH, 3-butenyl) and glucobrassicanapin (4-pentenyl). There were no differences in the types and concentrations of glucosinolates present in the terminal and lateral racemes of any variety, and only small differences between single- and double-low varieties. Concentrations of glucosinolates in floral tissues of crops grown with a little nitrogen were twice those of well-fertilised crops. There was no association between the number and distribution of pollen beetles on plants and the amounts or types of glucosinolates present in floral tissues.     

Could learning of pollen odours by honey bees (Apis mellifera) play a role in their foraging behaviour?

Abstract. The role of pollen odour cues in the foraging behaviour of honey bees (Apis mellifera L.) is poorly understood. Using classical conditioning of the proboscis extension response, in which bees learn to associate an odour with a sucrose reward, the present study tests whether odours of bee-collected pollen from the hive environment or odours of fresh pollen on the anthers of flowers could be used in pollen foraging. Honey bees efficiently learn odours from field-bean (Vicia faba) bee-collected pollen and oilseed-rape (Brassica napus) bee-collected pollen, hand-collected pollen, anthers and whole flowers, demonstrating that honey bees can learn pollen odours associatively in biologically realistic concentrations. Honey bees learn pollen odours of oilseed rape better than field bean and, although they generalize these two odours, they easily distinguish between them in discrimination tests, suggesting that pollen odours may be used in species recognition/discrimination. There is little evidence that honey bees can recognize whole flowers based on previous experience of bee-collected pollen odour. However, they generalize the odours of oilseed-rape anthers and whole flowers, suggesting that anther pollen in situ may play a more prominent role than bee-collected pollen in foraging behaviour.     

Effect of flower traits and hosts on the abundance of parasitoids in perennial multiple species wildflower strips sown within oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>) crops

Reducing the use of insecticides is an important issue for agriculture today. Sowing wildflower strips along field margins or within crops represents a promising tool to support natural enemy populations in agricultural landscapes and, thus, enhance conservation biological control. However, it is important to sow appropriate flower species that attract natural enemies efficiently. The presence of prey and hosts may also guide natural enemies to wildflower strips, potentially preventing them from migrating into adjacent crops. Here, we assessed how seven flower traits, along with the abundance of pollen beetles (Meligethes spp., Coleoptera: Nitidulidae) and true weevils (Ceutorhynchus spp., Coleoptera: Curculionidae), affect the density of parasitoids of these two coleopterans in wildflower strips sown in an oilseed rape field in Gembloux (Belgium). Only flower traits, not host (i.e. pollen beetles and true weevils) abundance, significantly affected the density of parasitoids. Flower colour, ultraviolet reflectance and nectar availability were the main drivers affecting parasitoids. These results demonstrate how parasitoids of oilseed rape pests react to flower cues under field conditions. Similar analyses on the pests and natural enemies of other crops are expected to help to develop perennial flower mixtures able to enhance biological control throughout a rotation system.     

The spatial dynamics of pollen beetles in relation to inflorescence growth stage of oilseed rape: implications for trap crop strategies

Semi‐field‐scale arrays of oilseed rape (Brassica napus L.) (Brassicaceae) plants were used to observe the development of distributions of pollen beetles (Meligethes aeneus Fabricius) (Coleoptera: Nitidulidae) in a simulated trap crop system where inflorescence growth stage alone was used to manipulate the pest. Over two successive years, pairs of 1 m spaced square arrays of 100 glasshouse‐grown plants were placed 40 m apart in the field in May, and were subject to natural infestation by pollen beetles. The test plot of each pair had a simulated trap crop, with an outer row of plants at early flowering stage intended to protect more susceptible inner plants at late bud stage, and the control plot had all plants at the late‐bud stage, simulating a standard crop situation. Pollen beetles were counted daily on each plant for 10–13 days. The spatio‐temporal development of plot infestation was analysed in relation to the distribution of racemes in bud and raceme in flowers using Spatial Analysis by Distance IndicEs (SADIE), and tests of edge and centre distribution. Inflorescence growth stage characteristics were shown to be important in determining the spatial distributions of pollen beetles. In control plots, the numbers of racemes in bud and in flower were never edge or centre distributed. In test plots, racemes in flower were always edge distributed, and racemes in bud began edge distributed and became centre distributed. Pollen beetle numbers were usually spatially associated with the abundance of racemes in bud and/or in flower. In control plots, pollen beetles were neither edge nor centre distributed, but in test plots they maintained a significant edge distribution for 7–10 days. At the end of the experiments, females were more centre distributed in the test plots than males, and were more closely associated with racemes with buds, whereas males were more associated with racemes with flowers. In early flowering stage plants, the number of racemes in flowers were a good indicator of the abundance of racemes in buds, but this relationship was lost as flowering progressed. Although flowering racemes provide strong cues for immigrating pollen beetles, the abundance of buds may be a more important determinant of residence time, particularly for females, and is therefore a critical determinant of trap crop effectiveness.     

Round goby Neogobius melanostomus attraction to conspecific and heterospecific egg odours

In this study, cannibalistic and predatory responses of juvenile Neogobius melanostomus (either fed or deprived of food and caught during the reproductive summer or non-reproductive winter season) to water washings of fertilized eggs from conspecifics (N. melanostomus), heterospecifics (rainbow trout Oncorhynchus mykiss) and control lake water were tested. Fed juveniles (summer season) spent significantly more time on the side of the flume containing stimuli from conspecific eggs over control water, while significantly preferring control water to heterospecific stimuli. In addition, fish showed a significant preference towards stimuli from conspecific eggs over heterospecific stimuli by associating near conspecific egg odours 1·5 times longer [Correction added after online publication 28 April 2011, text changed]. Fish that were deprived of food, regardless of whether caught during the summer or winter season, did not show any preference towards either conspecific or heterospecific stimuli. These findings show that conspecific egg odours attract fed juveniles and that there is a potential to lure fish to odour traps as a means of control.     

Pheromonal Recognition of Females Takes Precedence over the Chromatic Cue in Male Iberian Wall Lizards Podarcis hispanica

Sex recognition is based on colour signals in many species of lizards. However, olfactory stimuli are also clearly involved, and many species might rely more on chemoreception. We aimed to examine whether colour pattern or odours, or both, are used in sex recognition and which cues elicit courtship of females by males of the lizard Podarcis hispanica . We experimentally manipulated the coloration and odour of female P. hispanica , thereby creating groups with all combinations between coloration and odour of males and females. Using data from staged encounters, we compared the responses of resident males to manipulated and unmanipulated individuals (males and females). Responding males reacted significantly more aggressively to female intruders with male odours, independently of their coloration. Nevertheless, coloration seemed to be important in long-distance sex recognition since, in the first minutes, females painted as females received a lower number of aggressive responses. Both colour and odour were important in eliciting male courtship. However, females painted as females and with female odours were preferentially courted. Comparisons with unmanipulated male and female intruders agreed with these expectations. Therefore, at close range, odoriferous cues seem to be more important than colour patterns in sex recognition, but female coloration is also useful at long range to deter the aggressive response of males and to elicit courtship in conjunction with odours.     

Plant scents modify innate colour preference in foraging swallowtail butterflies

Flower-visiting insects exhibit innate preferences for particular colours. A previous study demonstrated that naive Papilio xuthus females prefer yellow and red, whereas males are more attracted to blue. Here, we demonstrate that the innate colour preference can be modified by olfactory stimuli in a sexually dimorphic manner. Naive P. xuthus were presented with four coloured discs: blue, green, yellow and red. The innate colour preference (i.e. the colour first landed on) of the majority of individuals was blue. When scent from essential oils of either orange flower or lily was introduced to the room, females’ tendency to select the red disc increased. Scents of lavender and flowering potted Hibiscus rosa-sinensis, however, were less effective. Interestingly, the odour of the non-flowering larval host plant, Citrus unshiu, shifted the preference to green in females. In males, however, all plant scents were less effective than in females, such that blue was always the most favoured colour. These observations indicate that interactions between visual and olfactory cues play a more prominent role in females.     

Colour choice behaviour in the pollen beetle Meligethes aeneus (Coleoptera: Nitidulidae)

The pollen beetle Meligethes aeneus Fabricius (Coleoptera, Nitidulidae), a pest of oilseed rape (Brassica napus), is known to respond to coloured stimuli; however, current understanding of the underlying mechanisms of colour choice in this species is limited. In the present study, physiological and behavioural experiments are conducted to determine the response of the pollen beetle to colours in the field. Spectral sensitivity is measured in 10 animals using the electroretinogram technique. Light flashes (100 ms) at varied wavelengths (340–650 nm, 10‐nm steps) and at different light intensities are applied to the eye after dark adaptation. In behavioural experiments in the field, 100 water traps of varying colours (from yellow to green to blue with varying amounts of white and black added, and with known spectral reflectance) are set out on a bare soil field in May 2008. The mean spectral sensitivity curve of M. aeneus peaks at 520 nm; however, a model template fitted to the long wavelength tail of the observed curve reveals a peak at approximately 540 nm (green). A secondary sensitivity peak is observed in the ultraviolet (UV) range (370 nm). A total of 2482 pollen beetles are captured in the coloured traps. The results show that the pollen beetles' preference for yellow over other colours can be modelled as a colour opponent mechanism (green versus blue); however, further experiments are needed to specify responses to colours with higher UV reflectance. These findings may be used to optimize trap colours for monitoring to help develop integrated pest management strategies for pollen beetle control.     

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

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

Honey-bee-mediated Infection of Pollen Beetle (Meligethes aeneus Fab.) by the Insect-pathogenic Fungus, Metarhizium anisopliae

Pollen beetles (Meligethes aeneus) are pests that feed and oviposit in the buds and flowers of oilseed rape. Honey-bees foraging from a hive fitted with an inoculum dispenser at the entrance effectively delivered dry conidia of the entomogenous fungus, Metarhizium anisopliae , to the flowers of oilseed rape in caged field plots. In both winter- and spring-sown rape experiments, a greater mortality of pollen beetles occurred in treated plots than in control plots. The mortality (61% on winter rape, 100% on spring rape) was greatest during peak flowering, when the feeding activity of both bees and beetles from the flowers was maximal, providing optimal conditions for inoculum dissemination and infection. Conidial sporulation occurred on a significant proportion of the dead pollen beetles. There was no evidence of any adverse effect on the honey-bee colonies.     

Discrimination of family‐specific odours by juvenile coho salmon: roles of learning and odour concentration*

Free‐swimming coho salmon fry Oncorhynchus kisutch of some families showed preference (relative to the behaviour of naïve sibs) for the odours of similarly aged non‐sibs to whom they had been exposed during the post‐hatch (alevin) stage and the early free‐swimming (fry) stage, but not the embryo (egg) stage, indicating that odour‐learning had occurred during the later developmental periods. Recognition (i.e. preference) of sib‐ pecific odours was evident after a month, and in one case 5 months, of separation from those odours. Thus, young salmon incubating in their gravel nests in streams appear to have the capacity to learn the chemical characteristics of conspecifics and retain this memory for at least several months without reinforcement. However, in addition to the general attractiveness of sibs and familiar non‐sibs over unfamiliar non‐sibs, some non‐sibs were consistently more attractive than others. Preference between two different non‐sib odours could be reversed by changing their relative concentrations, indicating that relative attractiveness is a function of both familiarity and odour concentration. Therefore, although juvenile coho salmon learn, remember, and are subsequently attracted by sib‐specific odours in early life, familiar odours are not always preferred over unfamiliar conspecific odours. Preference in dyadic assays is therefore an insensitive measure of recognition.     

Brief exposure to the odour of a parasitized male alters the subsequent mate odour responses of female mice

Parasites can affect mate choice, with females preferentially selecting parasite-free males. Prior exposure to, or experience with, males has also been suggested to influence mate responses. Here, we examined the effects of an immediate, brief (1 or 15 min) pre-exposure to the urinary and associated odours of either an uninfected male house mouse,Mus domesticus , or a male infected with the natural murine nematode parasite, Heligomosomoides polygyrus, on the subsequent responses of female mice to the odours of infected and uninfected males. Using an odour preference test we found that females displayed a marked overall preference for the odours of uninfected males. Pre-exposure to the odours of a parasitized male decreased female preference for nonparasitized males and increased female preference for parasitized males. This ‘prior male’ effect was evident for both the total odour preference and initial odour choice. Females pre-exposed to the odour of a parasitized male displayed decreased choosiness and made no particular initial choice of a male, while still maintaining an overall preference for the odours of uninfected males. These shifts in initial choice and odour preference were not directly associated with either female stress responses or stress-related odour cues of the males. Our findings show that female mice can distinguish between parasitized and nonparasitized males on the basis of odour and suggest that a brief exposure to the odours of infected males influences females' immediate odour preferences, and their subsequent preference for and choice of males. This rapid, infection-associated ‘prior male’ effect may contribute to the reported variations in female mate preference and choice. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour      

Distant olfactory response of the cabbage seed weevil, Ceutorhynchus assimilis, to oilseed rape odour in the field

Abstract. The olfactory orientation of adult cabbage seed weevils (Ceutorhynchus assimilis Payk.) from a distance of 20 m to extracts of oilseed rape (Brassica napus L.) was studied in the field using mark–release–recapture experiments. Male and female weevils dispersed upwind from the centre of a circular array of traps baited with Oilseed rape extracts. In the absence of host-plant odour, female weevils dispersed randomly, and male weevils downwind. Percentage recapture of weevils was significantly greater in traps baited with rape flower than rape leaf extracts. Significantly fewer weevils were recaptured in unbaited traps compared to traps baited with rape flower or leaf extracts. We suggest that seed weevils can use odour-mediated upwind anemotaxis to locate their host-plants from distances of at least 20 m.