Systemically-induced response of cabbage plants against a specialist herbivore, Pieris brassicae

Summary. Plant responses to herbivory might directly affect the herbivore (“direct” defences) or might benefit the plant by promoting the effectiveness of natural antagonists of the herbivores (“indirect” defences). Brussels sprouts attacked by Pieris brassicae larvae release volatiles that attract a natural antagonist of the herbivores, the parasitoid Cotesia glomerata, to the damaged plant. In a previous study, we observed that feeding by caterpillars on the lower leaves of the plant triggers the systemic release of volatiles detectable by the parasitoids from upper leaves of the same plant.?The role of these systemically induced volatiles as indirect defence and the dynamics of their emission were investigated in wind-tunnel dual choice tests with C. glomerata. The systemically induced emission of volatiles varied depending on leaf age and on plant age. Systemic induction affected parasitoid effectiveness, as induced plants could be more easily located by parasitoids than non-induced ones.?The role of the systemic induction as a direct defence was investigated through behavioural and feeding tests with P. brassicae. In dual choice assays, 1st instar larvae preferred to feed and fed more on systemically induced than on non-induced leaves. In single choice assays, the leaf area consumed by caterpillars was larger on systemically induced leaves than on non-induced control leaves. However, caterpillars fed on systemically induced leaves attained the same weight as those feeding on non-induced controls. In addition, P. brassicae pupae whose larvae were fed on systemically induced leaves had longer developmental times than those of larvae fed on non-induced leaves. Adult oviposition behavior was not influenced by systemic induction.?We conclude that systemically induced responses in cabbage might reduce P. brassicae fitness both directly, by affecting their development and feeding behavior and indirectly by making caterpillars and pupae more vulnerable to attack by carnivores. The occurrence of a possible relationship between direct and indirect defence is discussed. Received 24 January 2001; accepted 3 May 2001.     

Variability in herbivore-induced odour emissions among maize cultivars and their wild ancestors (teosinte)

Summary. Maize plants respond to caterpillar feeding with the release of relatively large amounts of specific volatiles, which are known to serve as cues for parasitoids to locate their host. Little is known about the genetic variability in such herbivore-induced plant signals and about how the emissions in cultivated plants compare to those of their wild relatives. For this reason we compared the total quantity and the qualitative composition of the odour blend among eleven maize cultivars and five wild Zea (Poaceae) species (teosinte), as well as among the offspring of eight Zea mays mexicana plants from a single population. Young plants were induced to release volatiles by mechanically damaging the leaves and applying oral secretions of Spodoptera littoralis (Lepidoptera: Noctuidae) caterpillars to the wounded sites. Volatiles were collected 7 h after treatment and subsequently analysed by gas chromatography. The total amounts of volatiles released were significantly different among maize cultivars as well as among the teosintes. Moreover, striking differences were found in the composition of the induced odour blends. Caryophyllene, for instance, was released by some, but not all varieties and teosintes, and the ratios among monoterpenes and sesquiterpenes varied considerably. The offspring of different mother plants of the Z. m. mexicana population showed some variation in the total amounts that they released, but the composition of the odour blend was very consistent within the population of this teosinte species. We discuss the ecological significance of these findings in terms of specificity and reliability of induced plant signals for parasitoids.     

Species-specific responses of herbivores to within-plant and environmentally mediated between-plant variability in plant chemistry

Allocation of resources to growth and defense against herbivores crucially affects plant competitiveness and survival, resulting in a specific distribution of assimilates and defense compounds within plant individuals. Additionally, plants rarely experience stable environmental conditions, and adaptations to abiotic and biotic stresses may involve shifts in resistance to herbivores. We studied the allocation of phytochemicals in Brassica oleracea (Brussels sprouts) due to leaf age, drought stress and herbivore damage and assessed effects on two lepidopteran herbivores differing in diet breadth: the generalist Spodoptera littoralis and the specialist Pieris brassicae. Glucosinolates as secondary defense compounds and total nitrogen and carbon were quantified and linked to plant palatability, i.e., herbivore feeding preference. Herbivore responses were highly species-specific and partially related to changes in phytochemicals. Spodoptera littoralis preferred middle-aged leaves with intermediate levels of glucosinolates and nitrogen over young, glucosinolate and nitrogen rich leaves, as well as over old leaves, poor in glucosinolates and nitrogen. In contrast, P. brassicae preferred young leaves. Both species preferred severely drought-stressed plants to the well-watered control, although analyzed glucosinolate concentrations did not differ. Both S. littoralis and P. brassicae feeding induced an increase of indole glucosinolate levels, which may explain a reduced consumption of damaged plants detected for S. littoralis but not for P. brassicae. By revealing distinct, sometimes contrasting responses of two insect herbivores to within-plant and stress-mediated intraspecific variation in phytochemistry of B. oleracea, this study emphasizes the need to consider specific herbivore responses to understand and predict the interactions between herbivores and variable plants.     

Behavioral and electroantennographic responsiveness of adult butterflies of six nymphalid species to food-derived volatiles

While adults of most species of butterflies visit flowers for food (nectar), those of certain species can feed on rotting food (e.g., exuded tree sap and rotting fruits). These food sources considerably differ in odor; flowers emit species-specific scents and rotting-food odors are characterized by fermentation products. To determine whether adult butterflies have different olfactory performances adaptive to their food selections, we examined the following responses of naïve adults of six nymphalid species, namely, Nymphalis xanthomelas, Kaniska canace, Vanessa indica, Cynthia cardui, Hypolimnas bolina, and Argyreus hyperbius: the proboscis extension reflex (PER) was evaluated as the behavioral criterion of preference for scent compounds, and the electroantennographic (EAG) response was assessed as the sensory criterion of antennal perception. All of these species are nectarivorous, but N. xanthomelas, K. canace, and V. indica also forage rotting food. The PERs to 52 tested compounds were in good agreement with the food habits of these butterflies. The six species showed high PERs to several aromatic compounds widely found in flower scents. Fermentation products such as ethanol and acetic acid elicited intermediate to high responses from the three species feeding on rotting foods. The other three species showed low PERs to these compounds, and acetic acid strongly inhibited the PER-eliciting activities of the other compounds. These results demonstrate that the olfactory preference for fermentation products is a characteristic of the rotting-food feeders. The spectrum of EAG responses to 37 tested compounds was relatively similar among the six species. Particular acyclic aliphatic compounds, including green leaf volatiles, elicited high EAG responses from all the species. Despite a high PER-eliciting activity, most aromatic compounds induced intermediate EAG responses; however, ethanol and acetic acid evoked very weak responses. The results suggest that in food selection, adult butterflies use a specific olfactory system to perceive these major food-derived volatiles.     

New insights in analysing parasitoid attracting synomones: early volatile emission and use of stir bar sorptive extraction

Summary. It is well known that feeding by Pieris brassicae caterpillars on cabbage leaves triggers the release of volatiles that attract natural antagonists such as the parasitoid Cotesia glomerata. The temporal dynamics in the emissions of parasitoid attracting volatiles has never been elucidated in this system. In a time course experiment, caterpillar infested leaves attracted the parasitoid within one hour after infestation. At such an early stage of infestation, as much as fifty percent of the parasitoids flew towards the infested plant in a wind tunnel bioassay, while only five percent flew towards the non-infested control plant. Three hours after infestation and later, the response to the volatiles from the infested plant reached its maximum and then continued at a constantly high level for the remaining 14 hours of the experiment. Chemical analyses of volatiles collected from infested leaves at short time intervals during the first 24 hours identified a total of ten compounds, comprising green leaf volatiles, terpenoids, and a nitrile. Significant increase of emission within the first 5 hours following initial herbivory was detected for (Z)-3-hexen-1-ol, (Z)-3-hexen-1-yl acetate, cineole and benzylcyanide. Subsequently, a coupled bioassay-chemical analysis procedure was developed allowing for testing and analyzing the same sample for future identification of the bioactive compounds. This was achieved by using stir bar sorptive extraction for the analysis of solvent extracts of caterpillar-damaged leaves.     

Multiplexed species-specific PCR protocol to discriminate four N. Atlantic Calanus species, with an mtCOI gene tree for ten Calanus species

Accurate species identification is the cornerstone of any ecological study - yet this fundamental step is not always possible for marine zooplankton. Routine species' identification, especially of juvenile andlarval stages, is difficult for Calanus species (Copepoda; Calanoida) in the N. Atlantic Ocean, where two or three species may co-occur. A rapid, simple, and inexpensive molecularly based protocol to identify individual copepods of any life stage has beendeveloped. This protocol will routinely identify four Calanus species in the N. Atlantic and will help to accurately understand the role of each species in coastal and open ocean ecosystems. The DNA sequence of a 633 bp region of the mitochondrial cytochrome oxidase I (mtCOI) was determined for ten Calanus species: C. australis (Brodsky, 1959), C. chilensis (Brodsky, 1959), C. finmarchicus (Gunnerus, 1770), C. glacialis (Jaschnov, 1955), C. helgolandicus (Claus, 1863), C. hyperboreus (Krøyer, 1838), C. marshallae (Frost, 1974), C. pacificus (Brodsky, 1948), C. simillimus (Giesbrecht, 1902), and C. sinicus (Brodsky, 1965). MtCOI sequences were used to design species-specific oligonucleotide primers for C. finmarchicus, C. glacialis, C. helgolandicus, and C. hyperboreus and to optimize a competitive, multiplexed, species-specific PCR (SS-PCR) protocol to discriminate the four species. This corrects and improves a previously published protocol for three Calanus species (Bucklin et al. 1999: Hydrobiologia 401:239-254), unambiguously identifying individual copepods and copepodites from diverse geographic regions of the four species' ranges. In order to further examine the pattern of mtCOI evolution within Calanus (an important consideration for molecular systematic characters), consensus mtCOI sequences were used to reconstruct phylogenetic relationships among the ten species; the mtCOI gene tree agreed with morphological and molecular (based on mt16S rRNA) phylogenies, except that the affiliation of C. sinicus could not be resolved.     

Emission of oilseed rape volatiles after pollen beetle infestation; behavioural and electrophysiological responses in the parasitoid <Emphasis Type="Italic">Phradis morionellus</Emphasis>

Summary. The pollen beetle, Meligethes aeneus, is an important pest of oilseed rape, Brassica napus. Larvae of this species feed only in the buds and flowers of Brassicaceae. One important natural enemy of this beetle is the parasitoid Phradis morionellus that attacks larvae in buds and flowers and also feeds on the flowers. The preferences for odours of non-infested and infested rape were tested for both starved and fed parasitoids in Y-tube olfactometer experiments. The volatile blend released from pollen beetle-infested and non-infested flowering rape and from pollen beetle larvae was identified and quantified. Gas chromatography-electroantennodetection analyses were performed with female P. morionellus. Parasitoids in both treatment groups preferred infested rape, but the proportion of responding female P. morionellus was significantly lower for the group that was starved. Six of the 20 volatiles identified were released at higher rates from infested rape than from non-infested. None of these compounds was found in pollen beetle larvae headspace. P. morionellus antennae detected both major and minor components in the volatile blend. The volatiles released at a significantly higher rate from infested rape and detected by P. morionellus antennae were (Z)-3-hexenylacetate, (Z)-3-hexenol, 3-butenyl isothiocyanate and (E,E)-α-farnesene.     

Potential parasitoid attractants,volatile composition throughout a bark beetle attack

Several hymenopteran parasitoids prey on mature bark beetle larvae (Scolytidae), concealed under the bark of conifers. According to previous results, certain oxygenated monoterpenes are crucial in host location. However, synthetic baits tested in laboratory bioassays are not quite as attractive to the parasitoids as the natural volatile bouquet from conifers containing susceptible bark beetle larvae. The aim of the current study was to identify the "missing components" in previously tested synthetic baits. Volatiles were sampled throughout bark beetle attacks (Ips typographus) on Norway spruce logs (Picea abies), and analyzed (qualitatively and quantitatively) by coupled gas chromatography-mass spectrometry (GC-MS). In addition, a pilot test was done employing the relatively new technique, fast GC-surface acoustic wave detector (SAW), zNose^TM, described in the methods. Quantitative differences in volatile composition were found between infested and uninfested logs, and between different stages of the bark beetle attack. In addition, the volatile composition of logs with or without mature bark beetle larvae (susceptible to parasitism) was compared, isolating odor differences at this most suitable host stage. In general, the quantity of monoterpenes (the predominant phytochemical volatiles in conifers) was found to decrease with time after initial infestation. In contrast, certain oxygenated monoterpenes and benzenoid compounds increased as susceptible bark beetle larvae developed, which therefore could be candidates for improving parasitoid baits. To the best of our knowledge, this is the first report on long-term dynamics of the volatile composition of bark beetle infested P. abies. The results should be interpreted in the light of previous electrophysiological and behavioral investigations of larval parasitoids, and present a number of compounds that could improve synthetic parasitoid baits.     

Comparative electrophysiological analysis of plant odor perception in females of threePapilio species

Summary Antennae of femalePapilio butterflies perceive many volatile plant constituents with widely differing, constituent-specific sensitivities. We compared the responses of threePapilio species to volatiles from host and non-host plants to assess species-specificity and the degree of evolutionary conservatism in olfactory responses.Since previous studies had demonstrated that the polar constituents in odor fromDaucus carota stimulate oviposition behavior inPapilio polyxenes, we collected headspace volatiles fromD. carota, Pastinaca sativa (both Apiaceae) andArtemisia dracunculus (Asteraceae) and separated the polar fraction of these volatiles by gas chromatography. GC-coupled electroantennograms (GC-EAG) were recorded from the speciesPapilio polyxenes, P. machaon hippocrates andP. troilus. In addition, the responses of the three species to five compounds known as generally occurring constituents of plant odor were recorded. The relative sensitivities for these compounds were nearly identical in all threePapilio species. The response spectra to the separated plant volatiles also showed considerable similarities among the species.From the limited set of GC peaks evoking a response in one of the species, 64% (D. carota), 44% (P. sativa) and 29% (A. dracunculus) also evoked a response in both of the other species. The responses of the two closely related Apiaceae feeders (P. polyxenes, P. m. hippocrates) to volatiles fromD. carota were more similar to each other than was either to the response ofP. troilus, which feeds on Lauraceae. However, this was not true for the responses to volatiles fromP. sativa. The least congruence among the three species was found in the responses to volatiles fromA. dracunculus, a non-host for all of them. The differences and similarities found in the response profiles of the threePapilio species are discussed with respect to evolutionary adaptation to host odor versus evolutionary conservatism in adaptation of olfactory receptors.     

Lack of sequestration of host plant glucosinolates in <Emphasis Type="Italic">Pieris rapae</Emphasis> and <Emphasis Type="Italic">P. grarricae</Emphasis>

Summary. Sequestration of plant toxins in herbivores is often correlated with aposematic coloration and gregarious behaviour. Larvae of Pieris brassicae show these conspicuous morphological and behavioural characteristics and were thus suggested to sequester glucosinolates that are characteristic secondary metabolites of their host plants. P. rapaeare camouflaged and solitary, and are thus not expected to sequester. To test this hypothesis and to check the repeatabi-lity of a study that did report the presence of the glucosinolate sinigrin in P. brassicae, larvae were reared on three species of Brassicaceae (Sinapis alba, Brassica nigra and Barbarea stricta), and different leaf and insect samples were taken for glucosinolate analysis. The major host plant glucosinolates could only be found in traces or not at all in larval haemolymph, bled or starved larvae, faeces or pupae of both species or P. brassicae regurgitant. Haemolymph of both Pieris spp. was not rejected by the ant Myrmica rubra in dual-choice assays; the regurgitant of P. brassicae was rejected. This suggests the presence of compounds other than glucosinolates that might be sequestered in or produced by P. brassicae only. In faeces of both Pieris spp. a compound which yielded 4-hydroxybenzylcyanide (HBC) upon incubation with sulfatase was detected in high concentrations when larvae had been reared on S. alba. This compound may be derived from hydrolysis of sinalbin, the main glucosinolate of that plant. The unidentified HBC progenitor was apparently not sequestered in the two Pieris spp., and was not detected in faeces of larvae reared on B. nigra or B. stricta. Received 18 July 2002; accepted 11 September 2002.     

Weak defence in a tritrophic system: olfactory response to salicylaldehyde reflects prey specialization of potter wasps

Predatory arthropods are attracted to infochemicals emitted by their herbivore prey or by the prey’s host plants. We studied such a tritrophic system measuring the olfactory responses of three potter wasp species (Symmorphus murarius, Symmorphus gracilis, Discoelius zonalis, Hymenoptera: Eumeninae) to salicylaldehyde, sequestered as a defence compound by Chrysomela leaf beetle larvae when feeding on Salicaceae, and volatile organic compounds (VOCs) emitted by aspen (Populus tremula, Salicaceae). In electroantennographic recordings (EAG), the highly specialized S. murarius that almost exclusively feeds on larvae of Salicaceae-feeding Chrysomela species was more sensitive to salicylaldehyde than the less specialized S. gracilis, feeding on such Chrysomela species but also weevil larvae. In contrast the related D. zonalis, foraging for microlepidoptera caterpillars on various host plants, did not respond at all. Furthermore, the three wasp species responded differently to aspen VOCs in GC–MS/EAD measurements. These results indicate that the sense of smell of predatory potter wasps differs for prey and plant volatiles among related wasp species according to their degree of host specialization. The considerable differences in salicylaldehyde perception suggest that its originally defensive function has backfired as it is used by specialist potter wasps for prey location. This is an important clue on adaptive mechanisms of the highest trophic level of the well-studied evolutionary arms race among Chrysomela leaf beetles, their host plants and their enemies.     

Ecological,genetic, and morphological differences among three <Emphasis Type="Italic">Pavona</Emphasis> (Cnidaria: Anthozoa) species from the Pacific coast of Panama

Ecological, genetic, and morphological differences among three Panamanian Pacific Pavona species with strongly developed collines (Pavona varians, P. frondifera, and P. chiriquiensis) were examined. Ecological factors included geographical distributions of species, habitat preferences, interspecific interactions, reproductive ecology, and tolerance to bleaching. Genetic differences were based on the electrophoretic analysis of ten allozyme loci. Morphological analyses consisted of tissue coloration, colony morphology, and measurements and counts of ten macro- and micro-skeletal characters. P. varians, present on reefs or in coral communities, is the most widely distributed and shows considerable morphological variation. P. chiriquiensis, a recently described species, encrusts basalt rock and has little morphological variation. P. frondifera is a reef dweller with a compact foliose morphology. Tissue coloration varies from light to dark brown in P. varians, from pink to brown in P. frondifera, and from brick red to brown or silvery in Pavona chiriquiensis. Also, the white to silvery polyp mouths of the latter species are a diagnostic feature that allows an easy identification in the field. Aggressive dominance during short-term interspecific interactions were as follows: Pavona chiriquiensis>P. varians>P. frondifera. P. chiriquiensis and P. varians showed contrasting responses to sea warming during the 1997-1998 El Niño Southern Oscillation. Whereas entire P. chiriquiensis bleached and died within 4 weeks of exposure to 30-31°C, colonies of P. varians did so only on their upper surfaces. The response of P. frondifera to elevated temperatures was not observed because it is mainly present in the Gulf of Panama where coral bleaching was absent in 1997-1998. The genetic data indicated that P. chiriquiensis differed strongly from both P. varians and P. frondifera, with Nei's unbiased genetic distances of 0.434 and 0.379, respectively. A fixed difference between P. varians and P. frondifera, and P. chiriquiensis exists at the triose phosphate isomerase (TPI-2) locus. A nearly fixed difference between P. chiriquiensis and P. frondifera and between P. chiriquiensis and P. varians was found at the hexokinase (HK) locus. P. varians differed slightly from P. frondifera with Nei's unbiased genetic distance of 0.068. No fixed difference was found between P. varians and P. frondifera. There were strong differences between P. chiriquiensis and P. varians in spawning times and gamete characteristics. Spawning in P. varians and P. chiriquiensis is 12 h out of phase. Also, eggs of the former species are white to beige and positively buoyant whereas those of the latter species are dark green and neutrally to negatively buoyant. No reproductive data are yet available for P. frondifera. Calicular diameters are significantly greater in P. chiriquiensis than in the other two species. In contrast, corallum thickness is greater in P. varians and P. frondifera than in P. chiriquiensis. Canonical discriminant function analysis readily separated the three species.     

Oviposition stimulants and deterrents control acceptance ofAlliaria petiolata byPieris rapae andP. napi oleracea

Summary Differential acceptance of garlic mustard,Alliaria petiolata byPieris rapae L. andP. napi oleracea is explained by their differential sensitivities to oviposition stimulants and deterrents in the plant. Fractions containing the stimulants and deterrents were isolated by solvent partitioning between water and n-butanol and by open-column chromatography followed by HPLC.P. napi oleracea showed no preference when offered a choice ofA. petiolata or cabbage, but was strongly stimulated to oviposit by post-butanol water extracts ofA. petiolata. The most abundant glucosinolate in this extract was identified as sinigrin, which could explain the high degree of stimulatory activity.P. rapae preferred cabbage plants overA. petiolata, and the relatively low stimulatory activity was also associated with the glucosinolate-containing aqueous extract. However, this species was strongly stimulated by a fraction that contained small amounts of glucotropaeolin along with unknown compounds. Deterrents to both species were found in the butanol extract fromA. petiolata, andP. napi oleracea was more sensitive thanP. rapae to these deterrents. Some HPLC fractions from the BuOH extract were strongly deterrent toP. napi oleracea, but were inactive toP. rapae. The ecological significance of these behavioral differences between the twoPieris species is discussed.     

Feeding ecology of five fishes from the mid-slope micronekton community off southern Tasmania, Australia

We examine the feeding ecology of characteristic micronekton species inhabiting the mid-slope region (~700 to 1,500 m depths) off southern Tasmania. Five fishes, Diaphus danae, Hygophum hanseni, Lampanyctus australis (Myctophidae), Phosichthys argenteus (Phosichthyidae) and Chauliodus sloani (Chauliodontidae), were sampled by depth-stratified midwater trawling on a diel and seasonal basis. Overall, 74 prey taxa were identified from 2,132 stomachs. Euphausiids (mostly Euphausia spinifera and E. similis) and calanoid copepods (mostly Pleuromamma species) were the main prey of the three myctophids; P. argenteus ate fishes and decapods in addition to the euphausiids, while C. sloani ate only fishes. Copepods were less important in the diets of larger D. danae, L. australis and P. argenteus and were replaced by euphausiids in the myctophids and by fishes in P. argenteus. In autumn, when euphausiid biomass increased six-fold, all three myctophids and P. argenteus fed most intensively and consumed a high proportion of euphausiids. The three myctophids appeared to feed nocturnally. Differences in the timing and duration of feeding corresponded to differences in their spatio-temporal overlap with key prey. Daily rations of H. hanseni (1.93% dry-weight biomass) and L. australis (1.43%), estimated from data on stomach fullness, were typical for temperate myctophids and higher than that of the non-migratory P. argenteus (0.43%). The vertical flux of near-surface plankton production to the mesopelagic food web is based primarily on diel feeding in the upper water column (<500 m) rather than consumption of species that migrate seasonally into the deeper mesopelagic zone. Because species such as P. argenteus and C. sloani feed above the third trophic level, their predators, including the commercially important orange roughy, are feeding between levels four and five.     

Electrophysiological and behavioural responses of Aphidius ervi (Hymenoptera: Braconidae) to tomato plant volatiles

Flight responses of the aphid parasitoid Aphidius ervi to tomato volatiles have recently demonstrated that different plant stresses can lead to increases in attractiveness for this parasitoid. For example, infestation of tomato plants by the aphid Macrosiphum euphorbiae results in the overexpression of defensive genes, as well as the release of volatile compounds that attract aphid parasitoids. Here, we determine which of the induced compounds elicit a significant electrophysiological response from parasitoid antennae. Compounds shown to be detected at the antennal level were then tested at a range of doses in a wind tunnel assay. A significant electroantennogram response was demonstrated for three compounds, (8S,9R)-(E)-caryophyllene, methyl salicylate, and (Z)-3-hexen-1-ol, over four concentrations. These compounds proved to be significantly attractive in the wind tunnel at a rate not always proportionally dependent upon the dose. The practical implications of these findings are discussed in the framework of sustainable control for pest aphids in agriculture.     

Maize field odorscape during the oviposition flight of the European corn borer

Most crop pests find a suitable host through chemical cues released from plants, but little is known about the odorscape encountered by host-seeking gravid females under natural, outdoor conditions. In this field study, the volatile organic compound (VOC) composition of maize (Zea mays, L.), a host for the European corn borer (ECB) (Ostrinia nubilalis Hüb.) was characterized during the oviposition flight and compared with a forest odorscape. VOCs from maize fields and the forest atmosphere were collected by solid phase microextraction and characterized by gas chromatography-mass spectrometry. The electroantennographic (EAG) response of female ECB antennae to candidate VOCs was tested. Analyses revealed clear differences between the maize field and the forest odorscapes, mainly composed of ubiquitous VOCs but in specific ratios. The maize field odorscape is more complex than the forest odorscape for maize found 18 VOCs but only eight in the forest. Both biotopes shared seven VOCs—green leaf volatiles (GLV), monoterpènes (MT) and homoterpenes. In addition, we found in the forest a distinctive sesquiterpene (SQT) identified as isoledene. The highest EAG responses were elicited by two GLVs and a MT shared by the two biotopes. SQT elicited weak EAG responses, except β-farnesene, only found in the maize field odorscape. Our results suggest that the two biotopes produce specific chemical signatures that insects may use as host cues. To the best of our knowledge this paper is the first report on the maize odorscapes under field conditions. The putative role of the VOCs in host plant detection and selection is discussed.     

Molecular systematic and phylogenetic assessment of 34 calanoid copepod species of the Calanidae and Clausocalanidae

DNA sequences for a 639 bp region of mitochondrial cytochrome oxidase I (mtCOI) were determined for 34 species of ten genera in two families of calanoid copepods, including: Calanoides, Cosmocalanus, Meoscalanus, Nannocalanus, Neocalanus, and Undinula (family Calanidae); and Clausocalanus, Ctenocalanus, Drepanopus, and Pseudocalanus (family Clausocalanidae). MtCOI gene sequences proved to be diagnostic molecular systematic characters for accurate identification and discrimination of the species. Levels of mtCOI variation within species (range: 1-4%) were significantly less than those between species (9-25%). Higher levels of intraspecific variation (>2%) usually resulted from comparisons between ecologically distinct or geographically isolated populations. MtCOI sequence variation resolved evolutionary relationships among species of Clausocalanus, Neocalanus, and Pseudocalanus, although there was evidence of saturation at some variable sites. Phylogenetic relationships among 11 copepod genera (adding Calanus to the list above) were reconstructed using a 660 bp region of nuclear small-subunit 18S rRNA, a slowly evolving gene that showed no variability within a species and differed by <1-6% among the genera. The 18S rRNA molecular phylogeny was consistent with the accepted limits of the Calanidae and Clausocalanidae and clearly resolved relationships among genera within each family. This molecular systematic and phylogenetic study was part of the ZooGene project, an international partnership to create a DNA sequence database as a tool for uniform, molecularly based species identification of planktonic calanoid copepods and euphausiids.     

Size-related trophodynamic changes in three target fish species recovering from intensive trawling

The consequences for food-web structure of protecting marine communities from trawling have been little explored. We investigated the Gulf of Castellammare, northwest Sicily, where a trawl ban initiated in 1989 has led to great increases in the biomass of some target species. Using stable isotopes of carbon and nitrogen, our objectives were to see if mean trophic level and omnivory had increased after the ban in three of the most common fishery-target species, namely, the anglerfish Lophius budegassa, Mediterranean hake Merluccius merluccius, and red mullet Mullus barbatus. We compared size data from before and after the fishery closure, but we also compared recently derived data from outside and inside the closed area. In all three species '¹³C and '¹⁵N were found to differ markedly depending on fish size. '¹³C was found to decrease in L. budegassa and Merluccius merluccius but increase in Mullus barbatus with increasing fish length. '¹⁵N increased in all three species, and this was thought to reflect feeding at increasingly higher trophic levels during the animals' lives. Mean length, and therefore '¹⁵N-derived trophic level, increased after the trawl ban only in the anglerfish L. budegassa. Based on '¹³C and '¹⁵N data, none of the species became more or less omnivorous after the ban. After 9 years of no trawling, increases in numerical abundance were not accompanied by substantial size-related trophodynamic shifts in any of the three species of fish studied.     

Electrophysiological responses and field attraction of the grey corn weevil, Tanymecus (Episomecus) dilaticollis Gyllenhal (Coleoptera: Curculionidae) to synthetic plant volatiles

The grey corn weevil, Tanymecus (Episomecus) dilaticollis, is an economically important polyphagous pest in Eastern and Central Europe. The present research is the first published electrophysiological study of olfactory sensitivity of adult T. dilaticollis to plant volatiles. The electrophysiological responses of male and female antennae were recorded to 20 synthetic volatile organic compounds (VOCs), previously identified to be released from the preferred host plants, maize and sunflower. Antennae of both sexes responded to a wide range of volatiles from different chemical classes: fatty acid derivatives, aromatic compounds and terpenoids. Two green leaf volatiles, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol, and one terpenoid, (±)-linalool, elicited the strongest electrophysiological responses by male and female antennae. Relatively high electrophysiological activity by T. dilaticollis antennae was also recorded to benzaldehyde, eugenol and caryophyllene. In the field, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol, when presented individually, caught significantly more male and female T. dilaticollis than unbaited controls. These compounds could be developed into an attractive trap for the detection and monitoring of T. dilaticollis.     

Biological patterns and near-bottom population characteristics of two pasiphaeid decapod crustacean species, Pasiphaea sivado and P. multidentata, in the north-western Mediterranean Sea

Biological aspects of the only two epi-/mesopelagic pasiphaeid species distributed along the continental marginof the north-western Mediterranean Sea were studied over the period 1991-1994. The annual reproductive cycle, reproductive output, sex and size distributions by depth (near-bottom distribution), and growth were analysed using 4156 specimens of Pasiphaea sivado (Risso, 1816) and 5491 specimens of P. multidentata Esmark, 1866. The reproductive cycle in the shallower-dwelling species, P. sivado, was nearly continuous, in contrast to the reproductive cycle in P. multidentata, which was markedly seasonal in nature. Both species exhibited similar size and population structure patterns with depth, with individual size increasing along the bottom depth gradient. The shallower-dwelling species had a significantly higher reproductive output level, in terms of the relative number of eggs, than the deeper-dwelling species. Biological trends for each of the species have been compared. The comparative results for these two congeneric species revealed that the biological parameters analysed were closely linked to their population structure. The slight differences in the depth range inhabited by each species have been considered the main environmental factor affecting the life histories of these two species. The data presented supported the hypothesis that depth exerted an important influence on the biological patterns of deep-water species, with the seasonality of reproductive processes increasing in deeper-dwelling species.