Host recognition in the pupal parasitoid Trichopria drosophilae: a morpho-functional approach

The diapriid wasp Trichopria drosophilae Perkins (Hymenoptera: Diapriidae) attacks and develops in puparia of the common fruit fly, Drosophila melanogaster Meigen (Diptera: Drosophilidae). Host recognition of T. drosophilae was studied using both a morphological and behavioural approach. Scanning and electron microscopical observations of female parasitoid antennae showed the presence of two types of sensilla, which we named MGS1 and MGS2. The former are present on the ventral side of both the apical (A11) and sub-apical (A12) antennomeres, while the latter occur only on A12. Ultrastructural features suggest a gustatory function for these sensilla. Arena bioassays using intact or antennaectomised females and intact host puparia showed that MGS2 are necessary for achieving host acceptance. Further bioassays, where the host's anterior spiracles were covered with wax, led to a very low level of host acceptance. We suggest that the secretion produced by glands associated with the anterior spiracles act as a contact kairomone, which has to be perceived by MGS2 in order to elicit host recognition. The removal of both the female apical antennomeres (A12) led to the failure of the parasitoid to recognize its host.     

Oviposition behaviour in Lygus rugulipennis: a morpho-functional study

Before oviposition, a Lygus rugulipennis (Poppius) (Heteroptera: Miridae) female explores the substrate with her mouthparts, performing ‘probing’ behaviour which eventually leads to a choice of oviposition site. In this study, morphological and behavioural observations were carried out in order to define the mechanisms through which female bugs recognise suitable oviposition sites. The morphological study was conducted using electron‐microscopy techniques, while the behavioural experiments aimed at the temporary (using ZnSO₄) or permanent (i.e., cauterization of the rostrum and ablation of both stylets and labium) inactivation of sensory structures located in the labium tip and in the stylets, which are probably involved in oviposition site selection. The tip of the labium of L. rugulipennis females is characterised by the presence of 11–12 uniporous gustatory sensilla which are innervated by 3–6 sensory neurones. One aporous mechanoreceptor is located ventrally on both areas of the labium tip. Other aporous mechanosensilla are found more proximally on the labium. ZnSO₄ labium tip treatment did not affect oviposition site selection, while stylet amputations, as well as rostrum cauterisations, resulted in almost complete oviposition failure. Labium tip‐ablated females oviposited similarly to control females. These results suggest that the sensory neurones associated with the stylets are involved in the location of the oviposition site. Uniporous gustatory sensilla on the labium tip are not involved in the final steps of the oviposition behaviour. However, they seem to play a role in assessing the suitability of the substrate as a food source.     

Anatomy of the antennal dorsal organ in female of Neodryinus typhlocybae (Hymenoptera: Dryinidae): A peculiar sensory structure possibly involved in perception of host vibration

Neodryinus typhlocybae (Hymenoptera: Dryinidae) is a natural enemy of the planthopper Metcalfa pruinosa, which was introduced from North America into Europe and has become established in various regions as a pest species. Vibrational signals play a crucial role in the communication of M. pruinosa, which appears to be exploited by N. typhlocybae. Scanning and transmission electron microscopy have shown that the antennae of N. typhlocybae females have peculiar and complex sensory structures: deep longitudinal grooves that house long sensilla trichodea, termed here “Antennal Dorsal Organs.” Such structures were not present on male antennae. These sensilla extend for the length of the grooves, without contact with the groove cuticle. Their hair shaft is empty and aporous, and inserted into a specialized socket, underneath which there is a cuticular ampulla‐like chamber. Each sensillum is associated with two sensory neurons: one terminates at the proximal end of the dendritic sheath; the other continues into the sensillum sinus and is enclosed in the dendritic sheath. This second sensory neuron then enters the ampulla‐like chamber through the circular opening, and then terminates with a conspicuous tubular body at the shaft base. The possible involvement of this peculiar structure in the context of host recognition mechanism is discussed. J. Morphol. 277:128–137, 2016. © 2015 Wiley Periodicals, Inc.     

Host recognition cues of the granary weevil parasitoid Lariophagus distinguendus

Host recognition was examined in Lariophagus distinguendus (Förster), a parasitoid of larvae of the granary weevil Sitophilus granarius (L.) that live endophytically in wheat grains. On encountering a grain infested with S. granarius, females of L. distinguendus behave in a set sequence. First they show antennal drumming on the grain, second they tap with the tip of the abdomen on the grain surface, third they drill into the grain and then insert their ovipositor. Bioassays revealed that drumming and drilling was stimulated by non-volatile chemicals present on the grain-host complex. Host faeces and herbivore damaged grain material stimulated the most activity, followed by artificially damaged grain, and healthy grain. This is the first report on non-volatile chemicals released from herbivore-damaged seeds as signals for foraging parasitoids. Volatile chemicals from the faeces alone were not active. Experiments on the use of physical cues revealed that the presence of a three-dimensional structure increased the response towards chemicals from the faeces. The shape (ovoid or rectangular solid) and colour (brown or white) of the structure had no impact. Thus, physical cues alone were insufficient to stimulate host recognition behaviour, but acted by increasing the response towards the chemical stimuli.     

Ultrastructural identification of the antennal gland complement in Siagona europaea Dejean 1826, a myrmecophagous carabid beetle

We examined antennal exocrine glands in adults of a myrmecophagous carabid beetle, Siagona europaea Dejean 1826 (Coleoptera, Carabidae), by light and electron microscopy and we identified two types of integumentary glands. The first type includes glands formed by three cells (a secretory cell, an intercalary cell and a duct cell) known as class 3 of Noirot and Quennedey (1991). The secretory cell has several large multivesicular electron‐lucent bodies, indicating a glycoprotein product associated with lipids. We hypothesize that this secretion protects the surface of antennae and sensilla from wear. The second group of glands includes unicellular glands known as oenocytes (class 2 of Noirot and Quennedey, 1991), which secrete epicuticular hydrocarbons through epidermal cells.     

Quantitative analysis of antennal mosaic generation in Drosophila melanogaster by the MARCM system

Mosaics have been used in Drosophila to study development and to generate mutant structures when a mutant allele is homozygous lethal. New approaches of directed somatic recombination based on FRT/FLP methods, have increased mosaicism rates but likewise multiple clones in the same individual appeared more frequently. Production of single clones could be essential for developmental studies; however, for cell-autonomous gene function studies only the presence of homozygous cells for the target recessive allele is relevant. Herein, we report the number and extension of antennal mosaics generated by the MARCM system at different ages. This information is directed to obtain the appropriated mosaic type for the intended application. By applying heat shock at 10 different developmental stages from 0-12 h to 6-7 days after egg laying, more than 50% of mosaics were obtained from 5,028 adults. Single recombinant clones appeared mainly at early stages while massive recombinant areas were observed with late treatments.     

Peptidoglycan recognition protein LF: a negative regulator of Drosophila immunity

Peptidoglycan recognition proteins (PGRPs) play important roles in the innate immune defence. Each PGRP detects a distinct subset of peptidoglycans and initiate immune signalling or enzymatic degradation of peptidoglycans. Here we characterize one of the 13 Drosophila PGRPs, PGRP-LF. PGRP-LF is membrane bound and has its two PGRP domains, z and w, localized outside the cell. Our data demonstrate that the z-and w-domain differ in their affinities to peptidoglycan. The z-domain has affinity to several groups of peptidoglycans while the w-domain only recognizes peptidoglycan from Escherichia coli. In addition, we observed that overexpression of PGRP-LF in Drosophila melanogaster Schneider 2 cells (S2 cells) promotes aggregation of cells. Furthermore, following immune stimulation of S2 cells overexpressing PGRP-LF, we noticed a reduced up-regulation of expression of antimicrobial peptide genes, in consonance with an immune suppressive role for PGRP-LF.     

Odor-mediated patch choice in the parasitoid Venturia canescens: temporal decision dynamics

Parasitoids foraging for hosts in a heterogeneous environment would greatly benefit if they could decide already from a distance in which areas search for resources would be most profitable and to avoid areas of low fitness returns. Interestingly, the temporal dynamics of the decision process in parasitoid patch choice have rarely been investigated. In a Y-tube olfactometer, we tested whether thelytokous and arrhenotokous females of the parasitoid wasp Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) respond to differences in cues indicating the quality of a host-containing patch and choose more profitable patches. Special attention was given to the time it took females to make their choices (patch choice time) when differences in patch quality were either qualitative (absence vs. presence of hosts and kairomone) or quantitative (various concentrations of hosts and kairomone, and presence of competitors). We found that both thelytokous and arrhenotokous wasps only chose the higher-quality patch based on odor cues when the difference was qualitative. When patches differed only with respect to the number of hosts, or the presence or absence of competing female parasitoids, no significant preference could be found in females of either strain of the parasitoid. In contrast, both the time until females reached the junction of the Y-tube olfactometer (response time) and the time until females decided for either patch (decision time) varied with parasitoid strain and odor treatment. Thelytokous wasps were faster than arrhenotokous wasps in their response time and in their decision time. However, females of both strains responded faster with increasing number of total hosts releasing kairomone. Yet, decision time for patches did not significantly vary as a function of patch quality offered to Venturia wasps.     

Geographical variation in resistance of the parasitoid Asobara tabids against encapsulation by Drosophila melanoqaster larvae: the mechanism explored

Abstract. The braconid parasitoid Asobara tabida Nees attacks larvae of several Drosophila species in fermenting substrates. Northwestern and central European populations of the parasitoid attack mainly D.subobscura Collin. Southern European parasitoids attack mainly D.melanogaster Meigen. Larvae of this last species can defend themselves against parasitoids by encapsulating the parasitoid egg. Parasitoids from southern European populations are better able to resist encapsulation of their eggs than their northwestern and central European conspecifics. The eggs of southern European parasitoids appear to have a 'sticky' egg chorion. As a result of this 'stickiness' the eggs become embedded in host tissue where they are not completely covered by the host's blood cells. This leads to, at most, partial encapsulation of the egg. Parasitoid larvae can escape from partially closed capsules.     

Spatial and temporal relationships between cell death and tissue overgrowth in imaginal wing disks of the Drosophila mutant I(3)C43hs1

The temperature-sensitive mutant l(3)c43^hs1 is lethal at the restrictive temperature late in the last larval instar and has wing disks that show excessive growth when larvae are reared at 25°C. Such mutant disks give rise to defective wings showing duplications and deficiencies. Abnormal folding patterns are localized to the region between the wing pouch and the area where adepithelial cells are found; the disks retain an epithelial morphology. Apoptotic cell death is distributed throughout the wing disks without any obvious concentration of dead cells in a specific area. Cell death is seen as early as 12 hr after a shift to the restrictive temperature. Temperature shift experiments also show that cell death precedes the onset of overgrowth, but since the spatial distribution of death is not localized to the regions of abnormal folds, it is unlikely that cell death and overgrowth are causally related.     

蒙古寒蝉成、若虫触角形态及若虫龄期判别研究（半翅目：蝉科）

对蒙古寒蝉若虫与成虫的触角感器进行了研究,并结合不同龄期若虫触角的形态特征,采用Dyar法则与“头宽—龄期”的回归曲线验证,初步断定蒙古寒蝉若虫具有4个龄期。不同龄期若虫与成虫的触角形态差异显著,触角感器的种类、数量和分布也明显不同,其差异或与若虫和成虫的生态位不同密切相关。     

Host preference and survival in selected lines of a Drosophila parasitoid,Asobara tabida

Host selection behaviour of parasitoids has important fitness consequences, if hosts of different quality are available. Here the host selection behaviour, the ability to distinguish between hosts differing in their suitability, of the Drosophila parasitizing wasp Asobara tabida was studied. Females from five lines selected for higher survival in the encapsulating host species D. melanogaster ( 16 ) were compared with females from control lines. Females from all five selected lines more readily accepted the encapsulating host species for oviposition when offered together with a nonencapsulating host species than females from the control lines. We found no evidence for pleiotropic effects and suggest that host selection behaviour evolved parallel to the ability to escape encapsulation in the hosts. Our results also suggest that given the appropriate selection pressures, host selection behaviour can quickly evolve in parasitoids, enabling them to adapt fast to changing circumstances.     

Host specificity of the egg pupal parasitoid Fopius arisanus (Hymenoptera: Braconidae) in La Reunion

Fopius arisanus is a polyphagous parasitoid of Tephritidae, which has been recently introduced to La Réunion Island as part of a classical biological control programme. We carried out laboratory experiments to assess the host specificity of this parasitoid, initially reared on Bactrocera zonata, and then offered for parasitization the eight local tephritid pest species. Naive or experienced parasitoid females were given tephritid eggs in no choice tests. Fopius arisanus females parasitize all fly species but parasitism varies with host species. No adult wasps emerge from Bactrocera cucurbitae and the survival of this species is only slightly affected by parasitism. Dissections show that the late instars of this fly may eliminate the parasitoid by encapsulation. When developing on Ceratitis capitata, Ceratitis rosa, Dacus ciliatus, Dacus demmerezi, and Neoceratitis cyanescens, parasitoid survival rate ranges from 10 to 25%. Bactrocera zonata and Ceratitis catoirii are the best hosts, yielding parasitoid survival rates of more than 70% with no premature mortality. The egg-larval mortality of C. capitata, C. rosa, D. ciliatus, and N. cyanescens, and the pupal mortality of D. demmerezi, are significantly increased by parasitism. The size of emerging adults is affected by host species and is correlated to pupal weight. Bactrocera zonata would be a favorable host to support routine colonization of F. arisanus for mass production of this parasitoid.     

Life history of Spalangia gemina Boucek (Hymenoptera: Pteromalidae), a fast-breeding microhymenopteran pupal parasitoid of muscoid flies

Laboratory studies at 25.5 degrees C and 70-75% r.h. demonstrated that the average daily survival rate for females of the parasitoid Spalangia gemina Boucek (Hymenoptera: Pteromalidae) was 0.907 over a 20-day period, with 50% mortality in 17.3 days. Mean immature developmental time of S.gemina was 22 days and the population growth rate was c. 40-fold per generation. The females successfully parasitized 1-, 2- and 3-day-old pupae of Musca domestica L. and parasitism of 98-100% was obtained at parasitoid to host ratios of 1:0.5-2.5. Parasitism of 81-87% was obtained at parasitoid to host ratios of 1:10. The male to female sex ratio was 1:1.59. S.gemina appears to have advantages over other Spalangia spp., which have immature development times of 28-32 days under comparable conditions, for the biological control of Muscidae.     

Variability and genetic components of innate fruit odour recognition in a parasitoid of Drosophila

Abstract This study aims to investigate the genetic variability of olfactory responses to odours from the host habitat, involved in host selection by insect parasitoids. The probing response of females to fruit and non-fruit odours was studied in Leptopilina boulardi Barbotin et al . (Hymenoptera, Figitidae), a parasitoid of the frugivorous Drosophila melanogaster larvae. Responses of two strains (from Nasrallah, Tunisia, and from Brazzaville, Congo) and their hybrids were compared. Probing is a searching mode by repeated ovipositor insertions into the breeding substrate to locate digging larvae. Most Brazzaville females exhibited innate ovipositor probing into agar in response to the fruit aromas (banana, orange, pear, prickly pear), whereas this was rare in the Nasrallah strain. These four fruits are all breeding sites for D. melanogaster, whereas mushroom is not. Mushroom odour did not trigger any probing response in either strain. In addition, the latency of the probing response was shorter in Brazzaville females. Results from hybrid females (F1 and Backcross) showed that both probability to probe in response to a fruit aroma and latency of the response were strongly heritable. Regarding the probability to probe, there was an apparent complete dominance of Nasrallah characteristics, whereas additive inheritance was found in the Brazzaville lineage. The contrast between reciprocal F1 hybrids suggested a strong interaction with non-chromosomal factors. It is suggested that a cytoplasmic factor in the Nasrallah strain decreases females' propensity to probe in response to significant odours. The latency of probing appeared to be under a chromosomal influence. Thus, this study identifies a strong genetic component in the responses to odours from the host habitat. It also provides information on the genetic basis of the two characters studied.     

Larval chemosensory projections and invasion of adult afferents in the antennal lobe of Drosophila

We have studied the fate of olfactory afferents during metamorphic transformation of Drosophila melanogaster. Intracellular labeling of afferents from larval head chemosensilla suggests that the larval antennal lobe may be an olfactory target, whereas tritocerebral and suboesophageal centers are likely targets of gustatory sensilla. Application of monoclonal antibody 22C10 shows that the larval antennal nerve is the precursor of the adult antennal nerve and is used as a centripetal pathway for the adult afferents. Likely guidance cues are larval olfactory afferents that persist during early metamorphosis. P[GAL4] enhancer trap lines are introduced as efficient markers to follow the establishment of adult sensory projections. β-Galactosidase and the bovine TAU protein were used as reporter proteins, and their expression patterns are compared. P[GAL4] lines MT14 and KL116 demonstrate that adult antennal afferents have arrived in the antennal lobe 24 h after pupariation and extend to the contralateral lobe 6 h later. Line MT14 expresses GAL4 mostly in basiconic sensilla and in certain trichoid sensilla, whereas KL116 is specific for trichoid and a small subset of basiconic sensilla. In the antennal lobe, largely complementary subsets of glomeruli are labeled by the two lines, in agreement with the observation that particular types of sensilla project to particular target glomeruli. © 1997 John Wiley & Sons, Inc. J Neurobiol 32: 281–297, 1997.     

Mechanism of mate detection in parasitoid wasps: sound and vibratory cues change with the developmental progress of future mates inside host pupal cases

Insects including parasitoid wasps use acoustic and vibratory signals in the context of sexual communication, mate recognition, courtship and mating. Males of the parasitoid wasp Pimpla disparis Viereck (Hymenoptera: Ichneumonidae) detect insect host pupae parasitized by a conspecific female, learn their location, visit them repeatedly and remain on or near them when the prospective mate nears emergence. In the present study, the acoustic and vibratory cues that males exploit to detect the presence and track the developmental progress of a future mate inside a host pupal case are investigated. Responses are acquired from developing parasitoids (DePa) by airborne sound and laser Doppler vibrometer recordings, after gently stimulating each of 20 wax moth host pupae with a paintbrush on days 1–23 post parasitism. Sound and vibratory cues produced by DePa are detectable from day 7 onward and relate mostly to spinning movements. Parameters of sound and vibratory cues (amplitude, dominant frequency, upper limit of frequency band) change significantly over time and thus could ‘inform’ a visiting adult male about the stage of development of DePa. Adult males antennating a parasitized pupa and flying around it also induce vibrations, which in turn may inform DePa about the presence of a male. There is no experimental evidence for true signalling and rapid information exchange between DePa and adult males. Delaying reply signals may help DePa avoid attacks by illicit receivers of such signals, including female (hyper)parasitoids and invertebrate predators.     

Genome scans of variation and adaptive change: extended analysis of a candidate locus close to the phantom gene region in Drosophila melanogaster

Nucleotide variation in populations originating from the recent range expansion of a species should reflect their adaptation to new habitats as well as their demographic history. A survey of nucleotide variation at 109 noncoding X-chromosome fragments in a European population of Drosophila melanogaster allowed identifying some candidates to have been recently affected by positive selection. Adaptive changes leave a spatial differential footprint that can be used to discriminate among candidates by extending their study to neighboring regions. Here, we surveyed variation at an approximately 190-kb region spanning a locus exhibiting a significantly skewed frequency spectrum. A stretch of approximately 12 kb with reduced variation was detected within a continuously sequenced region that included the focal fragment. Moreover, the regions flanking this stretch exhibited an excess of high-frequency derived variants. Application of maximum likelihood ratio and goodness-of-fit tests suggested that the pattern of variation detected at the studied region (at cytological bands 17C-17D) might have been shaped by a recent selective change, most probably at or around the phantom gene that encodes CYP306A1, a cytochrome P450 enzyme in the ecdysteroidogenic pathway.