Behavioural,ecological and genetic evidence confirm the occurrence of host‐associated differentiation in goldenrod gall‐midges

Host‐associated differentiation (HAD) is considered a step towards ecological speciation and an important mechanism promoting diversification in phytophagous insects. Although the number of documented cases of HAD is increasing, these still represent only a small fraction of species and feeding guilds among phytophagous insects, and most reports are based on a single type of evidence. Here we employ a comprehensive approach to present behavioural, morphological, ecological and genetic evidence for the occurrence of HAD in the gall midge Dasineura folliculi (Diptera: Cecidomyiidae) on two sympatric species of goldenrods (Solidago rugosa and S. gigantea). Controlled experiments revealed assortative mating and strong oviposition fidelity for the natal‐host species. Analysis of mitochondrial DNA showed an amount of genetic divergence between the two host‐associated populations compatible with cryptic species rather than host races. Lower levels of within‐host genetic divergence, gall development and natural‐enemy attack in the S. gigantea population suggest this is the derived host.     

Oviposition preferences of the Hessian fly and their consequences for the survival and reproductive potential of offspring

1. The relationship between oviposition preference and offspring performance was investigated experimentally for the Hessian fly Mayetiola destructor (Say) using 10 grass genotypes that represented five different genera and six different species. Oviposition preferences were quantified in a choice test using arrays of 200–400 plants. Offspring performance was estimated by recording survival during three phases of the insect–plant association and by measuring wing length, a correlate of adult reproductive potential. Density effects were examined for all offspring variables, and were taken into account when offspring performance was compared across the 10 grasses. 2. Egg counts from the oviposition choice test revealed a consistent ranking of plants: 18ITSN triticale > Otane bread wheat = Caldwell bread wheat = Stacy bread wheat > 3424 bread wheat = PND durum wheat = Fleet barley = Valetta barley = Matua brome grass > Awapuni oat. 3. Survival and adult wing length varied significantly on the 10 plant types. Survival was ranked: Otane bread wheat > 3424 bread wheat = Fleet barley = Matua brome grass > Caldwell bread wheat = Valetta barley > 18ITSN triticale = PND durum wheat = Stacy bread wheat > Awapuni oat. Adult male and female wing lengths were greatest on Otane, the bread wheat that also provided the best survival. 4. For Hessian flies on each of the 10 plant types, data on survival, wing length, and wing length–fecundity relationships were combined into a single fitness measure. When these fitness measures were compared with egg counts, no overall pattern emerged. On seven of the 10 plant types, there was a positive linear relationship between egg counts and offspring fitness. On the other three plant types, egg counts were high while fitness was low. Possible reasons why Hessian fly females lay eggs on plants that are poor hosts for their offspring are discussed.     

Carnivores and carotenoids are associated with adaptive behavioural divergence in a radiation of gall midges

1. Adaptive divergence in sympatry is supposed to be inhibited by the homogenizing role of gene flow. However, studies continue to uncover examples of sympatric divergence. In this study, two divergent phenotypes in a complex of four syntopic gall midge morphotypes [nominally Asteromyia carbonifera Osten Saken, Diptera: Cecidomyiidae: Alycaulini] are characterised. The first is a behavioural phenotype governing within‐host tissue preference and the second is a trait governing accessory‐gland carotenoid quality and quantity. 2. One gall morphotype (crescents) lay most of their eggs on mature tissue whereas the other three gall morphotypes oviposit only on young emerging leaves. Ecological maintenance of this divergent trait appears to be driven by enemy‐reduced space. That is, nearly 40% of the crescent morphotype galls that develop high on the plant are attacked by the egg parasitoid Platygaster solidaginis Ashmed, whereas those low on the plant are relatively protected. 3. All morphotypes contain carotenoids in their accessory glands, but the quality and quantity of these pigments differs significantly between the morphotypes and is positively associated with the probability of parasitism by P. solidaginis. 4. Larval salivary glands also contain carotenoids and the plant hormone abscisic acid, which in plants is synthesized from carotenoid precursors and is involved in regulating plant defences. This hormone may facilitate gall development and influence gall morphology. 5. Ecological fitness trade‐offs between carotenoids, parasitoid attack, and plant resistance may be fostering adaptive divergence in ovipositional phenotypes and sympatric speciation in this complex of gall midge morphotypes.     

Reproductive biology of Phillyrea angustifolia L.(Oleaceae) and effect of galling-insects on its reproductive output

The breeding system of Phillyrea angustifolia is examined in several Spanish populations, both in the southeastern Iberian Peninsula and in the Balearic IslandS. Hand-pollination experiments performed on one of the mainland and on one of the island populations demonstrated that the plant is functionally androdioeciouS. Pollen from males was several times more fertile than pollen from hermaphrodite flowers or self-pollen in the island population, but not so in the Iberian Peninsula. The interpretation of such a result is that androdioecy must have an adaptive advantage in an island system, where inbreeding depression is expected to be greater. Male individuals are much less common than hermaphrodites in all populations studied, thus supporting the prediction made by the models for the maintenance of androdioecy. Sex expression in an individual plant differs between populations. The frequency of sexes was purely bimodal in one population, whereas it was not so in another. Pollen from both male and hermaphrodite flowers appears to be morphologically different, although it remains unknown at what stage of the fertilization process the difference becomes functional. Fruitset in the hermaphrodite individuals was always less than 10%.Most fruits abort at an early stage and there is also a great flower abscission. Fruitset does not appear to be influenced by any of the plant traits describing size or fecundity. A high unpredictability of flowering in an individual was also observed in all populations.P.angustifolia is attacked by a cecidomyiid fly that induces the formation of galls in the ovaries (producing ‘deforme’ fruits which have been confused until recently with parthenocarpic fruits), thus reducing the plant reproductive output. The production of such galls (most of them in the hermaphrodite individuals but a small proportion also in males) is highly variable among plants and among populations, representing from 0 to 97% of the initiated fruitS. Such variability in gall incidence does not respond to variability in plant size, fecundity, distance to flowering conspecifics or time that flowers are available for fly oviposition. A high consistency in gall production was found in both the mainland and the island populations. The greater abundance of galls in the islands compared with the mainland populations is interpreted as a result of the higher temperatures in the former which might influence the activity period of the flies.     

Factors influencing the onset of egglaying in a cecidomyiid fly

Abstract The timing of the onset of egglaying in the Hessian fly, Mayetiola destructor (Say), is influenced by several exogenous and endogenous factors. Mated females initiated egglaying 48 h earlier than virgins and laid 3–4 times more eggs before dying. The effects of mating on the onset of egglaying were further modified by several other factors whose effects were measured in hours rather than days. Increases in ambient temperatures experienced by mated females shortened the post-mating pre-ovipositional transition phase; however, the effects of temperature were not linear throughout the temperature range. Age of females at the time of mating also influenced the onset of egglaying. Females that mated 1 , 2 and 3 h after eclosion spent 190 , 160 and 120 min in the post-mating transition phase, respectively. Effects of age were also observed when females were mated over the three calling periods that comprise the 3-day lifespan of the adult female. Linear regressions of transition times on time of mating had similar slopes for first and second day post-eclosion females but were offset such that first-day females mated late in the morning had similar transition times to second-day females mated early in the morning. By the third day post-eclosion, transition times showed no significant changes when females were mated throughout the calling period. Possible mechanisms underlying the effects of mating, temperature, and female age on post-mating pre-oviposition transition times are discussed.     

Revision of the European species of Omphale Haliday (Hymenoptera,Chalcidoidea, Eulophidae)

The European species of Omphale Haliday (Eulophidae: Entedoninae) are revised. The revision includes 37 species, of which eleven are newly described and the remaining 26 species are redescribed. The species are classified into six species groups, with six unplaced species. All species are fully diagnosed and thoroughly illustrated. Identification keys are provided for females and males. Two new morphological features to aid classification and identification are introduced: male genitalia and wing interference patterns (WIPs). The former has been used successfully in the classification of New World Omphale and the latter is used for the first time in a taxonomic revision. Male genitalia in Omphale have considerable interspecific variation, an unusual trait among chalcidoid Hymenoptera, and are demonstrated to be useful for classification of species and species-groups, and they also possess the only autapomorphy for Omphale. WIPs are useful to help separate some species, but cannot be used to define either the genus or species groups. Distributional data are compiled for each species and suggest a pan-european distribution for most species. Gall-midges are the known hosts for 14 species, and the absence of host overlap between species suggests that host specialization is a driving force for speciation. Several Omphale species are known only from females, or have a strong female biased sex ratio, suggesting thelytokous development. Apart from the 37 species included in this revision, the status for nine additional species (names) in species group aetius remain unsolved. For nomenclatorial stability, a neotype is designated for Eulophus lugens Nees (= Omphale lugens (Nees)). Elachestus obscurus Förster and Derostenus sulciscuta Thomson are transferred from Holcopelte to Omphale comb. n. Derostenus radialis Thomson and Achrysocharella americana Girault are synonymized with Omphale theana (Walker), and Omphale teresis Askew is synonymized with Omphale phruron (Walker), syn. n. The status of genus Pholema Graham is revised as it is removed from synonymy with Omphale and instead synonymized with Neochrysocharis Kurdjumov, syn. n., and the type species for Pholema, Pholema microstoma Graham, is transferred to Neochrysocharis, comb. n. Eugerium orbatum Szelényi, previously transferred to Omphale, is synonymized with Asecodes congruens (Nees), syn. n.     

Hessian fly Avirulence gene loss‐of‐function defeats plant resistance without compromising the larva's ability to induce a gall tissue

Plant pathogen effectors encoded by Avirulence (Avr) genes benefit the pathogen by promoting colonization and benefit plants that have a matching resistance (R) gene by constituting a signal that triggers resistance. The Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae), resembles a plant pathogen in showing R/Avr interactions. Because of these interactions, a wheat plant with the H13 resistance gene can be resistant or susceptible depending on the genotype of the larva that attacks the plant, being resistant if attack comes from a larva with a functional vH13 gene, but susceptible if attack comes from a larva with a non‐functional vH13 gene. In this study we asked: does this susceptible interaction involving plants with H13 look like susceptible interactions with plants lacking H13? Possibly, the H13 plant attacked by a larva with a non‐functional vH13 is induced to partial rather than complete resistance. Or the larva, lacking its vH13‐encoded effector, is compromised in its ability to induce susceptibility, which includes forcing the plant to create a gall nutritive tissue. Responses of epidermal cells to larval attack were explored using imaging techniques (light microscopy, scanning and transmission electron microscopy). Whole‐organism responses were investigated by measuring the growth of plants and larvae. No evidence was found for partial resistance responses by H13 plants or for a compromise in the ability of vH13 loss‐of‐function larvae to induce susceptibility. It appears that disrupting vH13 function is sufficient for overcoming the induced resistance mediated by the H13 gene.     

Intra‐plant heterogeneity influences the preference and performance of juveniles and adults of a gall midge

Abstract 1. Field studies were conducted to evaluate the preference and performance of a gall‐inducing midge (Harmandia tremulae) within the crown of trembling aspen (Populus tremuloides). Females did not select oviposition sites preferentially within leaves, but did lay preferentially on young leaves. 2. Larvae were the only life stage involved in gall site selection within leaves and in gall initiation and development. Gall size, which was positively related to survival, was highest for galls on mid veins that were located close to the petiole. However, one‐third of galls were located on lateral veins and most galls were not adjacent to the petiole, indicating that many larvae choose sub‐optimal gall initiation sites. 3. Gall density was positively associated with leaf length, and leaf length, was positively associated with gall size. However, gall density per leaf was not related to larval survival in galls. This latter result may be a result of an observed inverse relationship between gall size and gall density for similar‐sized leaves. 4. The results partially support the plant vigour and optimal plant module size hypotheses, which predict that galler fitness in successfully induced galls should be highest on large, fast‐growing plant modules. The lack of a strong preference‐performance link supports the confusion hypothesis, which predicts that oviposition and gall site selection may often be suboptimal in systems where galler lifespan is short. This study suggests that small‐scale variations in plant quality within leaves, can render gall site selection by juveniles as important as that previously reported for adult females.     

Description,phenology and biology of Zelostemma chionochloae Buhl sp. nov., a platygastrid parasitoid of Eucalyptodiplosis chionochloae (Diptera: Cecidomyiidae) in New Zealand

Abstract

A platygastrid wasp that feeds on the snow tussock flower midge in the inflorescences of New Zealand snow tussock grasses (Chionochloa spp.) is formally described. Zelostemma chionochloae is a specialist natural enemy of Eucalyptodiplosis chionochloae Kolesik which is the most ubiquitous and sophisticated seed predator of Chionochloa. Z. chionochloae is a koinobiont parasitoid and some larvae enter prolonged diapause inside their host for at least 2 years. Methods for adult rearing are described. The phenology of Z. chionochloae is highly synchronised with its host even after 2 years in diapause. Parasitism levels were found to differ between years and elevations, while sex ratios differed among years. Z chionochloae probably suffers inter‐specific competition with another host‐specific hymenopteran parasitoid (Gastrancistrus sp.) which also parasitises E. chionochloae.     

The effect of fluctuating asymmetry and leaf nutrients on gall abundance and survivorship

Fluctuating asymmetry (FA) is defined as small and random deviations from perfect symmetry in otherwise bilaterally symmetrical traits. Usually, plants under stress have a higher nutritional quality and are preferred by insect herbivores. Here, we examined the relationship among leaf FA of Bauhinia brevipes (Fabaceae), nutrients and the occurrence of Schizomyia macrocapillata (Cecidomyiidae: Diptera). We evaluated whether: (i) FA was related to S. macropillata abundance and survivorship; (ii) FA correlated with the levels of foliar nutrients (macro- and micronutrients, and organic carbon); and (iii) S. macropillata abundance was related to leaf nutrients. The results showed a positive relationship between FA and gall abundance. S. macrocapillata survivorship was five times higher in B. brevipes individuals with high FA levels, but this difference was not significant. The influence of nutrients in B. brevipes FA was negligible, except for foliar nitrogen, which was 7.3% higher in plants with low FA levels. The abundance of S. macrocapillata was negatively related to nitrogen levels, but gall survivorship was not affected by this nutrient. We found evidence for a causal relationship between FA and gall occurrence, since S. macropillata gall oviposition occurs in young undeveloped leaves, thus mature leaves show an increase in FA. Nonetheless, in this system, neither FA nor nitrogen influenced gall survivorship.