Hymenopteran parasitoids reared from European gall midges (Diptera, Cecidomyiidae).

We report the results of investigations 2010 through 2023 of hymenopteran parasitoids associated with gall midges in Europe. A total of 242 collections of gall midges were made, from each of which one to several parasitoid species emerged, resulting in ca. 200 recorded parasitoid species and 267 host-parasitoid interaction records. The parasitoid families involved were Eulophidae (63 species), Platygastridae (56 species), Torymidae (34 species), Pteromalidae (31 species), Ceraphronidae (5 species), Eupelmidae (4 species), Eurytomidae (2 species) and Encyrtidae (1 species). As many as 159 interactions are reported for the first time, significantly enlarging our knowledge of gall midge - parasitoid interactions on the species level. Even more interesting, 51 host records are for parasitoid species for which no host was previously known. Similarly, 28 species of gall midge are reported as host to named parasitoids for the first time. Additionally, 91 parasitoid records were the first for the country in question. Differences between the rearing methods applied and their suitability for recording species with contrasting life histories, are discussed.

Mycobiomes of two distinct clades of ambrosia gall midges (Diptera: Cecidomyiidae) are species-specific in larvae but similar in nutritive mycelia.

IMPORTANCE: Ambrosia gall midges are endophagous insect herbivores whose larvae live enclosed within a single gall for their entire development period. They may exhibit phytomycetophagy, a remarkable feeding mode that involves the consumption of plant biomass and mycelia of their cultivated gall symbionts. Thus, AGMs are ideal model organisms for studying the role of microorganisms in the evolution of host specificity in insects. However, compared to other fungus-farming insects, insect-fungus mutualism in AGMs has been neglected. Our study is the first to use DNA metabarcoding to characterize the complete mycobiome of the entire system of the gall-forming insects as we profiled gall surfaces, nutritive mycelia, and larvae. Interestingly, larval mycobiomes were significantly different from their nutritive mycelia, although Botryosphaeria dothidea dominated the nutritive mycelia, regardless of the evolutionary separation of the tribes studied. Therefore, we confirmed a long-time hypothesized paradigm for the important evolutionary association of this fungus with AGMs.

Post-glacial colonization of the Fennoscandian coast by a plant parasitic insect with an unusual life history.

Species that exhibit very peculiar ecological traits combined with limited dispersal ability pose a challenge to our understanding of ecological and evolutionary mechanisms. This is especially true when they have managed to spread over long distances, overcome physical barriers, and colonize large areas. Climate and landscape changes, trophic web relations, as well as life history all interact to shape migration routes and present-day species distributions and their population genetic structures. Here we analyzed the post-glacial colonization of northern Europe by the gall midge Contarinia vincetoxici, which is a monophagous parasite on the perennial herb White swallowwort (Vincetoxicum hirundinaria). This insect not only has a narrow feeding niche but also limited dispersal ability and an exceptionally long dormancy. Gall midge larvae (n = 329) were collected from 16 sites along its distribution range in Denmark, Sweden, and Finland. Using microsatellite loci and knowledge of the species and the regions' history, we investigated the role of landscape change, host plant distribution, insect population dynamics, and life history in shaping the population genetic structure of the insect. We devoted particular interest to the role of the insect's presumed poor dispersal capacity in combination with its exceptionally extended diapause. We found significant levels of local inbreeding (95% highest posterior density interval = 0.42-0.47), low-level within-population heterozygosity (mean H E = 0.45, range 0.20-0.61) with private alleles in all populations except two. We also found significant (p < .001) regional isolation-by-distance patterns, suggesting regularly recurring mainly short-distance dispersal. According to approximate Bayesian computations, C. vincetoxici appears to have colonized the study area via wind-aided flights from remote areas approximately 4600-700 years before present when the land has gradually risen above the sea level. Extremely long dormancy periods have allowed the species to "disperse in time", thereby aiding population persistence despite generally low census population sizes.

The Cuticular Hydrocarbons of Dasineura Oleae Show Differences Between Sex, Adult Age and Mating Status.

In insects, cuticular lipids prevent water loss and act as semiochemicals. Because of their ecological function, the profile change across the insects' sex and development offers insight into insect biology and possible tools for pest management. Here, the first work on cecidomyiid cuticular extracts is proposed considering Dasineura oleae (Diptera: Cecidomyiidae) males and females at different adult ages (0-12 h, 12-24 h, 24-36 h) and distinct sexual conditions (virgin and mated). A set of 49 compounds were recorded (12 alkanes, 1 monomethyl alkane, 11 fatty acids, 4 esters, 1 aldehyde, 1 allylbenzene, 1 amine, 1 flavonoid, 1 ketone, 1 phenol, 1 steradiene, 1 sterol, 1 terpene, 1 triterpene and 11 unknown compounds), and 18 of them showed significant differences between groups. Among alkanes, hexacosane (nC26) exhibited a decreasing trend from the youngest to the oldest females, while pentacosane (nC25) and nonacosane (nC29) showed a decreasing trend from 0 to 12 h to 12-24 h virgin females. In addition, nonadecane (nC19) was significantly more abundant in the youngest males compared to older males and females. The alkanes nC25, nC26 and nC29 have been reported to be age-related also in other dipterans, while nC19 has been described as gender-specific chemical cue for platygastrid parasitoids. Further behavioural trials and analyses are required to assign the specific ecological roles to the characterized compounds. Our results may contribute to develop new low-impact control strategies relying on the manipulation of D. oleae's chemical communication (e.g. disruption of mating or species recognition). HIGHLIGHTS: • Cuticular hydrocarbons are often involved in dipteran intraspecific communication. • We explored the cuticular profile of D. oleae at different age, sex, mating condition. • Five alkanes and one mono-methyl alkane showed differences among groups. • Linoleic acid is the most abundant compound in virgins, absent in mated insects. • Eleven compounds disappear in mated insects, but were present in all virgins.

Pest categorisation of Resseliella maxima.

The EFSA Panel on Plant Health performed a pest categorisation of Resseliella maxima (Diptera: Cecidomyiidae), the soybean gall midge, for the EU. This midge was first described in 2018 and is widespread in north-western United States. It is not listed in Annex II of Commission Implementing Regulation (EU) 2019/2072. Larvae feed on and develop in soybean (Glycine max, Fabaceae), and possibly in two other Fabaceae, sweet clover (Melilotus officinalis) and alfalfa/lucerne (Medicago sativa). Feeding damage results in dark brown or black areas on the stems which become weak and can break near the soil; heavy infestations can cause plant death. R. maxima adults live only a few days and adult females lay eggs within 24 h after emergence. Larvae of R. maxima overwinter in the soil as third instars in silken cocoons. The main natural dispersal stage is the adult, which can fly. Freshly cut host plants for animal feed contaminated with larvae provide a potential pathway for entry into the EU. However, there is great uncertainty as to whether such plants are imported from USA states where R. maxima occurs. Climatic conditions and host availability in central-western EU MS are favourable for outdoor establishment. Phytosanitary measures are available to reduce the likelihood of entry and spread. Except for the uncertainty concerning the likelihood of entry, R. maxima satisfies the other criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest.

Mechanisms of hydraulic conductivity in the leaf galls of Meunieriella sp. (Cecidomyiidae) in Avicennia schaueriana (Acanthaceae): does vascularization explain the preferred sites of induction?

Intralaminar galls of Meunieriella result from ground tissue proliferation in leaves of Avicennia schaueriana, a typical halophytic mangrove. We tested if the preferred sites of gall induction were the midribs and secondary veins (SV) at the basal leaf portion, where the galls were expected to be largest; and if the vascular system in galls and adjacent regions was altered to favour water supply in galls, thus increasing their growth. Gall induction sites and gall sizes were quantified according to leaf portions and regions. Anatomical and histometric analyses in vascular and ground tissues of galls and adjacent regions were compared to equivalent regions of non-galled leaves. The galls were largest at basal sites on leaves, the midrib and SV. More galls occurred on the apical portion of the leaf, and on the leaf blade and secondary vein regions. Changes in shape and vascular system area, number and diameter of vessel elements were detected in both galls and adjacent regions. Fewer and smaller-sized vessel elements were observed in regions proximal to the galls and inside them. Gall size is not related with preferred induction sites, which could be explained by factors such as thermal balance. Alterations in the vascular system indicate reduced hydraulic conductivity in the xylem in the proximal region and inside galls, leading to water leakage to gall parenchyma cells. This compensatory mechanism explains the expansion and proliferation of water storage and spongy parenchyma cells in the galls, explaining the higher growth in more vascularized regions.

Insect galls from the Serra da Bandeira (Barreiras, Western Bahia, Brazil) / Galhas de insetos da Serra da Bandeira (Barreiras, Oeste da Bahia, Brasil)

Abstract Understanding the diversity of insect galls is pivotal to the establishment of conservation planning in different Brazilian ecosystems. Here, we (1) characterize the insect galls found on plant host species, (2) identify the gall-inducing insects to the lowest possible taxonomic level, and (3) record the presence of gall-associated fauna. Our study was carried out monthly from August, 2017 to July, 2018 along nine trails in Serra da Bandeira, which is located in an area of Cerrado stricto sensu over a year. We found 48 distinct gall morphotypes, belonging to 12 botanical families. The most morphotypes commonly found were globoid (27.1%), lenticular (27.1%) and cylindrical (25%). The plant organs most affected by galling insects were the leaves (73%), followed by the stem (19%), flower (4%), inflorescence (2%) and fruits (2%). The host botanical families holding the highest number of galls were Fabaceae (26), Malpighiaceae (5) and Anacardiaceae, Combretaceae and Euphorbiaceae (3). We observed that the genus Copaifera Lindl. (Fabaceae) had the greatest diversity of gall morphotypes, harboring 19 morphotypes distributed in three species that occur sympatrically - Copaifera sabulicola J. Costa (Fabaceae), Copaifera depilis Dwyer (Fabaceae) and Copaifera luetzelburgii Harms (Fabaceae). For the first time in Brazil, we recorded the occurrence of galls on flowers of Manihot caerulescens (Euphorbiaceae), on inflorescences of Mimosa acutistipula (Fabaceae) and flower buds of Anacardium humile (Anacardiaceae), which were induced by Cecidomyiidae. Inducers of the order Diptera (Family Cecidomyiidae) were the most abundant, found in 14 morphotypes of galls. Regarding the associated fauna, we found insects primarily belonging the order Hymenoptera, and identified them as parasitoids. The information provided can be used highly by decision makers for conservation programs, as well as in other strategies for the conservation of biological diversity in the Brazilian Cerrado.

Resumo Compreender a diversidade de galhas de insetos é fundamental para o estabelecimento de planos de conservação em diferentes ecossistemas brasileiros. Aqui, nós (1) caracterizamos as galhas de insetos encontradas em espécies de plantas hospedeiras, (2) identificamos os insetos indutores de galhas até o nível taxonômico mais baixo possível e (3) registramos a presença de fauna associada a galhas. Nosso estudo foi realizado mensalmente de agosto de 2017 a julho de 2018 ao longo de nove trilhas na Serra da Bandeira, que está localizada em uma área de Cerrado stricto sensu ao longo de um ano. Encontramos 48 morfotipos distintos de galhas, pertencentes a 12 famílias botânicas. Os morfotipos mais comumente encontrados foram globoide (27,1%), lenticular (27,1%) e cilíndrico (25%). Os órgãos vegetais mais afetados pelos insetos galhadores foram as folhas (73%), seguidas do caule (19%), flor (4%), inflorescência (2%) e frutos (2%). As famílias botânicas hospedeiras com maior número de galhas foram Fabaceae (26), Malpighiaceae (5) e Anacardiaceae, Combretaceae e Euphorbiaceae (3). Observamos que o gênero Copaifera Lindl. (Fabaceae) apresentou a maior diversidade de morfotipos de galhas, abrigando 19 morfotipos distribuídos em três espécies que ocorrem simpatriicamente - Copaifera sabulicola J. Costa (Fabaceae), Copaifera depilis Dwyer (Fabaceae) e Copaifera luetzelburgii Harms (Fabaceae). Pela primeira vez no Brasil, registramos a ocorrência de galhas em flores de Manihot caerulescens (Euphorbiaceae), em inflorescências de Mimosa acutistipula (Fabaceae) e em botões florais de Anacardium humile (Anacardiaceae), induzidas por Cecidomyiidae. Indutores da ordem Diptera (Família Cecidomyiidae) foram os mais abundantes, encontrados em 14 morfotipos de galhas. Em relação à fauna associada, encontramos insetos pertencentes principalmente à ordem Hymenoptera, e os identificamos como parasitóides. As informações fornecidas podem ser utilizadas por tomadores de decisão para programas de conservação, bem como em outras estratégias para a conservação da diversidade biológica no Cerrado brasileiro.

Occurrence and characterization of insect galls in two reserves of the Peruvian Amazon / Ocorrência e caracterização de galhas de insetos em duas reservas da Amazônia peruana

Abstract An insect gall inventory was carried out in two reserves of the Peruvian Amazon, Allpahuayo-Mishana National Reserve and Quistococha Regional Reserve, both situated in Iquitos, northeastern Peru. Four vegetation types were surveyed between December, 2021 and December, 2022: terra firme forest, white-sand wet forest, and white-sand dry forest in Allpahuayo-Mishana National Reserve, and palm swamp forest in Quistococha Regional Reserve. Overall, we found 262 gall morphotypes, distributed across 75 host species representing 66 plant genera and 30 families. Fabaceae was the plant family with the greatest number of gall morphotypes (n = 48), followed by Calophyllaceae (n = 21) and Euphorbiaceae (n = 20). The plant genera that supported the highest diversity of galls were Caraipa (n = 17), Eschweilera (n = 16), Tapirira (n = 16), Micrandra (n = 14), and Iryanthera (n = 10). The plant species Tapirira guianensis (n = 16), Caraipa utilis (n = 14), Micrandra elata (n = 14), Eschweilera coriacea (n = 11), and Sloanea parvifructa (n = 10) exhibited the highest richness of galls. Among the host plants, C. utilis stands alone as the only species noted as both endemic to the Amazonian region and bearing a Vulnerable (VU) conservation status. The leaves were the most attacked organs (90% of all galls). Most morphotypes are glabrous (89%), green (67%), globoid (53%), and one-chambered (91%). We found galling insects belonging to the orders Diptera, Thysanoptera, Lepidoptera, and Hemiptera. The galling insects of Cecidomyiidae (Diptera) were the most common, inducing 22% of the gall morphotypes. In addition to the gallers, we also observed the presence of successors, cecidophages, and parasitoids. Among the sampled vegetation types, the terra firme forest presented the highest richness of gall morphotypes and host plant species. This is the first systematic inventory of insect galls in this part of the Peruvian Amazon.

Resumo Um inventário de galhas de insetos foi realizado em duas reservas da Amazônia peruana, Reserva Nacional Allpahuayo-Mishana e Reserva Regional Quistococha, ambas situadas em Iquitos, nordeste do Peru. Quatro tipos de vegetação foram pesquisados entre dezembro de 2021 e dezembro de 2022: floresta de terra firme, floresta úmida de areia branca e floresta seca de areia branca na Reserva Nacional Allpahuayo-Mishana, e floresta de pântano de palmeiras na Reserva Regional Quistococha. No total, encontramos 262 morfotipos de galhas, distribuídos em 75 espécies hospedeiras representando 66 gêneros de plantas e 30 famílias. Fabaceae foi a família de plantas com o maior número de morfotipos de galhas (n = 48), seguida por Calophyllaceae (n = 21) e Euphorbiaceae (n = 20). Os gêneros de plantas que apresentaram a maior diversidade de galhas foram Caraipa (n = 17), Eschweilera (n = 16), Tapirira (n = 16), Micrandra (n = 14) e Iryanthera (n = 10). As espécies de plantas Tapirira guianensis (n = 16), Caraipa utilis (n = 14), Micrandra elata (n = 14), Eschweilera coriacea (n = 11) e Sloanea parvifructa (n = 10) apresentaram a maior riqueza de galhas. Dentre as plantas hospedeiras, C. utilis destaca-se como a única espécie listada como endêmica da região amazônica e com um status de conservação Vulnerável (VU). As folhas foram os órgãos mais atacados (90% de todas as galhas). A maioria dos morfotipos é glabra (89%), verde (67%), globoide (53%) e possui apenas uma câmara interna (91%). Encontramos insetos galhadores pertencentes às ordens Diptera, Thysanoptera, Lepidoptera e Hemiptera. Os insetos galhadores da família Cecidomyiidae (Diptera) foram os mais comuns, induzindo 22% dos morfotipos de galhas. Além dos galhadores, também observamos a presença de sucessores, cecidófagos e parasitoides. Entre os tipos de vegetação amostrados, a floresta de terra firme apresentou a maior riqueza de morfotipos de galhas e espécies de plantas hospedeiras. Este é o primeiro inventário sistemático de galhas de insetos nesta região da Amazônia peruana.

From leaves to inflorescences: Gall induction of Iatrophobia brasiliensis Rübsaamen, 1915 on inflorescences of Manihot caerulescens Pohl (Euphorbiaceae) during the dry season

ABSTRACT Gall-inducing insects are highly specialized herbivores as they have the ability to control and redirect the development of host plants to obtain food and shelter. The distribution of galls on plants can be influenced by seasonality and phenological events, which determines the reproductive success of these insects. The species Manihot caerulescens Pohl (Euphorbiaceae) has a great diversity of gall-inducing insects in the Cerrado of Western Bahia. Our study aimed to (1) study the fauna associated with M. caerulescens Pohl (Euphorbiaceae) and (2) evaluate the phenological events of this host plant species. We performed gall collections between July 2018 and June 2020 and monitored 30 individuals of the host species to study the phenology in the Serra da Bandeira, Bahia, Brazil. The emerged insects in the laboratory were mounted on permanent slides and identified. We found galls on the stems, leaves and inflorescences. Stem galls were induced by lepidopterans (Alucitidae), and leaf and inflorescence galls by Iatrophobia brasiliensis Rübsaamen, 1915 (Cecidomyiidae). Further, we showed that I. brasiliensis preferentially induced galls on the leaves, however during the dry season, galls were induced on the inflorescences. Although the induction of galls on the leaves by I. brasiliensis has already been reported in the literature, here for the first time we record the presence of galls on the inflorescences induced by the same gall-inducing species. Our study constitutes an important contribution towards the knowledge of the insect-plant interaction between M. caerulescens and I. brasiliensis in the Cerrado of Bahia.

Entomogenous galls and their associated fauna in deciduous dry forest and woodland vegetation remnants (Caatinga) in the Northern Depressão Sertaneja, Brazil / Galhas entomógenas e sua fauna associada em remanescentes de floresta seca e vegetação lenhosa decídua (Caatinga) na Depressão Sertaneja Setentrional, Brasil

Abstract A survey and characterization of entomogenous galls and their associated fauna were carried out in six remnants of Caatinga in the Northern Depressão Sertaneja, in the states of Paraíba, Rio Grande do Norte and Ceará. We identified 41 gall morphotypes in 24 plant species, belonging to 12 botanical families. Fabaceae had 29.2% (n = 12) of the total number of gall morphotypes found. Leaves (61%) and stems (25%) were the most attacked organs. Most morphotypes are glabrous (75.6%), while only 24.4% exhibit trichomes. Most galls were induced by insects of the Cecidomyiidae family. The associated fauna comprised successors, cecidophages, tenants, and parasitoids. Sucessors were found in four morphotypes of galls and included spiders and four orders of insects: Hemiptera, Coleoptera, Lepidoptera, and Hymenoptera (Formicidae). The tenants were represented by Tanaostigmoides (Tanaostigmatidae). The parasitoids, found in 18 morphotypes (43.9%), were represented by six families of Hymenoptera. We recorded the occurrence of new types of galls in 12 species of host plants. These gall records are new references for the Northern Depressão Sertaneja in the studied states.

Resumo Realizou-se um levantamento e caracterização de galhas entomógenas e sua fauna associada em seis remanescentes de Caatinga presentes na Depressão Sertaneja Setentrional nos estados da Paraíba, Rio Grande do Norte e Ceará. Identificamos 41 morfotipos de galhas em 24 espécies de plantas, pertencentes a 12 famílias botânicas. Fabaceae abrigou 29,2% (n = 12) do total de morfotipos de galhas encontrados. As folhas (61%) e os caules (25%) foram os órgãos mais atacados. A maioria dos morfotipos é glabra (75,6%), enquanto apenas 24,4% exibiram tricomas. As galhas, em sua maioria, foram induzidas por insetos da família Cecidomyiidae. A fauna associada compreendeu sucessores, fungívoros, inquilinos e parasitoides. Os primeiros foram encontrados em quatro morfotipos de galhas, estando representados por aranhas e quatro ordens de insetos: Hemiptera, Coleoptera, Lepidoptera e Hymenoptera (Formicidae). Os inquilinos foram representados por Tanaostigmoides Ashmead, 1896 (Tanaostigmatidae). Já os parasitoides, encontrados em 18 morfotipos de galhas (43,9%), foram representados por seis famílias de Hymenoptera. Registramos a ocorrência de novos tipos de galhas em 12 espécies de plantas hospedeiras. Os registros de galhas também são novas referências para a Depressão Sertaneja Setentrional nos estados estudados.

Population Genetics of the Blueberry Gall Midge, Dasineura oxycoccana (Diptera: Cecidomyiidae), on Blueberry and Cranberry and Testing Invasion Scenarios.

We compared the population genetic structure between populations of the blueberry gall midge-Dasineura oxycoccana (Johnson) (Diptera: Cecidomyiidae)-from blueberry and cranberry and determined the genetic relationships among geographical subgroups by genotyping 632 individuals from 31 different populations from their native USA regions (New Jersey, Michigan, and Georgia) and from invaded Korean regions using 12 microsatellite loci. Our population genetic analyses showed a clear separation between the two host-associated D. oxycoccana populations from blueberry and cranberry. Using data from only the blueberry-associated D. oxycoccana populations, we identified five genetically isolated subgroups. An analysis of the approximate Bayesian computation suggests that the invasive D. oxycoccana population from Korea appears to have been introduced from an unsampled source population rather than directly from its native range. Our findings will allow for an easier identification of the source of D. oxycoccana into newly invaded regions, as well as to determine their association with blueberry and cranberry, which based on our results can be considered as two distinct species.

Components of the Female Sex Pheromone of the Newly-Described Canola Flower Midge, Contarinia brassicola.

The canola flower midge, Contarinia brassicola Sinclair (Diptera: Cecidomyiidae), is a newly-described species that induces galls on canola, Brassica napus Linnaeus and Brassica rapa Linnaeus (Brassicaceae). Identification of the sex pheromone of C. brassicola is essential to developing monitoring tools to elucidate the geographic range and hosts of this new pest, and the extent to which it threatens the $30 billion Canadian canola industry. The aim of this study was to identify and synthesize the female-produced sex pheromone of C. brassicola and demonstrate its effectiveness in attracting males to traps in the field. Two peaks were identified through GC-EAG analysis of female-produced volatiles which elicited electrophysiological responses in male antennae. These peaks were initially characterized through GC-MS and synthesis as 2,7-diacetoxynonane (major component) and 2-acetoxynonane (minor component), and the racemic compounds elicited EAG responses in male antennae. All four stereoisomers of 2,7-diacetoxynonane were synthesized and the naturally-produced compound was shown to be primarily the (2R,7S)-isomer by analysis on an enantioselective GC column, with a small amount of (2R,7R)-2,7-diacetoxynonane also present. The configuration of the minor component could not be determined because of the small amount present, but this was assumed to be (2R)-2-acetoxynonane by comparison with the configuration of the other two components. In field trials, none of the four stereoisomers of 2,7-diacetoxynonane, presented individually or as a racemic mixture, was attractive to male C. brassicola. However, dispensers loaded with a 10 µg:1 µg blend of (2R,7S)- and (2R,7R)-2,7-diacetoxynonane caught large numbers of male C. brassicola and significantly more than other blends tested. The addition of 0.5 µg of (2R)-2-acetoxynonane to this blend further increased the number of males caught. In future work, we will seek to identify the optimum trapping protocol for the application of the pheromone in monitoring and surveillance.

Anatomical profiles validate gall morphospecies under similar morphotypes.

Plant galls are generated by the stimuli of gall-inducing organisms on their hosts, creating gall morphotypes that vary in color, shape, size, and tissue organization. Herein, we propose to compare the structural features of gall morphotypes on the superhost Croton floribundus (Euphorbiaceae) in order to recognize gall morphospecies, i.e., galls with similar shapes but different internal structures. Non-galled leaves and galls were analyzed macroscopically, histologically, and histochemically for the detection of primary metabolites, and the results obtained were used for statistical analyses of similarity. Among the eight gall morphospecies, four are globoid, two are lenticular, one is fusiform and one is marginal leaf rolling. Stomatal differentiation and the occurrence of different types of trichomes were impaired in some gall morphospecies. Three patterns of organization of the ground system are recognized, ranging from the maintenance of mesophyll cells that differentiate into palisade and spongy cells dorsiventrally to the formation of a complex cortex with three morphofunctional layers. The marginal leaf rolling galls have the simplest anatomical structures, quite similar to those of the non-galled host leaf, while lenticular, globoid (types I to IV), and fusiform galls are anatomically more complex. Herein, we report on eight gall morphospecies occurring on C. floribundus, which are distinguished by morpho-anatomical attributes and show the disruption of the morphogenetic patterns of the host leaf toward the morphogenesis of unique gall features.

Functional Differentiation of Floral Color and Scent in Gall Midge Pollination: A Study of a Schisandraceae Plant.

Gall midges are among the most host-specific insects. Their interactions with plants likely date back to the Cretaceous period. Plants from at least seven families are involved in gall midge pollination; however, little is known about the pollination signals of gall midges. In this study, we used a Resseliella-Schisandra model to investigate the roles of floral scent and color in attracting gall midges. Field observations, behavioral bioassays via Y-tubes, and "flight box" experiments were performed. The results demonstrated that gall midges may be attracted by both floral scent and color and that two flower signals are more effective in promoting insect flower-landing than either alone. In the field, gall midges visited male flowers effectively at night but almost always visited female flowers during the day. Thus, during the Resseliella-Schisandra interactions, female flowers predominantly employed visual cues over scent to attract midges during the day; in contrast, olfactory cues were more functional for male flowers to export pollen in the dark. In this study, we first identified the roles of floral color and the functional differentiation of visual and olfactory cues during gall midge pollination.

Infestation of the gall midge Dasineura oleae provides first evidence of induced plant volatiles in olive leaves.

In this study, we present the first characterization of herbivore-induced plant volatiles (HIPVs) released from infested olive leaves. The gall midge Dasineura oleae is a specific pest of Olea europaea and endemic of the Mediterranean Basin, an area in which severe outbreaks currently occurred. Little is known about the damage caused by the pest and the relationship with its host. Since gall formation and larval feeding activity may lead to the release of specific plant volatile compounds, we investigated the volatile profiles emitted from infested plants compared with healthy plants under both laboratory and field conditions. Additionally, the volatiles emitted from mechanically damaged plants were considered. A blend of 12 volatiles was emitted from olive trees infested by D. oleae. Of these, ß-copaene, ß-ocimene, cosmene, unknown 1 and unknown 3 were found to be exclusively emitted in infested plants. The emission of germacrene-D, (E,E)-α-farnesene, and (Z,E)-α-farnesene, α-copaene, (E)-4,8-dimethylnona-1,3,7-triene, (E)-ß-guaiene and heptadecane significantly increased in infested trees. Linalool, ß-copaen-4-α-ol, ß-bourbonene, ß-cubebene, ß-elemene, ß-copaene and δ-amorphene were found only in the field trial and showed differences depending on the level of infestation and the plant stage. (Z)-3-Hexenol, (E)-4-oxohen-2-enal, and 2-(2-butoxyethoxy)-ethanol, were exclusively emitted from the leaves after mechanical damage. In a field trial in Italy, we also demonstrated spring synchronization between adults of D. oleae and O. europaea trees. Analyses of morphoanatomical malformations of gall leaves showed that tissue alterations occur at the spongy parenchyma causing an increase of the leaf blade thickness. We speculate that tissue alterations may lead to HIPV release, in turn potentially attracting D. oleae natural enemies.

Phenolic substances and cyanogenesis in galled and non-galled tissue of the fern species Microgramma vacciniifolia / Substâncias fenólicas e cianogênese em tecidos galhados e não galhados da samambaia Microgramma vacciniifolia

Galls, neo-formed plant structures that can occur in different organs, are generated by species-specific interaction with an inducing organism. Inducers manipulate the metabolism of its host. Microgramma vacciniifolia (Langsd. & Fisch.) Copel. is a Neotropical epiphytic fern that hosted two stem galls, one induced by a midge species (Diptera) and other by a micromoth species (Lepidoptera). The aim of this study was to evaluate the impact of these two gall-inducing insects on the biochemistry of phenolic acids and the cyanogenesis in galls, stems and leaves of M. vacciniifolia. High performance liquid chromatography (HPLC) indicated a total of 14 phenol derivatives, including caffeic and coumaric acid. Principal Coordinates Analysis (PCoA) of the phenolic substances indicated three groups consisting (1) non-galled stems and micromoth-induced galls; (2) midge-induced galls; (3) midgeinduced galls with parasitoids. Regarding the frequency of cyanogenesis assessed by the picrate paper test, the chi-squared test showed significant difference between fertile leaves (8.3%), sterile leaves (27.7%), non-galled stems (0%) and galls. Among galls, only the midge-induced galls analyzed were cyanogenic (15%). Our results indicated that the different gall-inducers (midge and micromoth) promote species-specific alterations to the phenolic substance composition of the host fern.

Galhas são estruturas vegetais neo-formadas que ocorrem em diferentes órgãos. Elas são geradas por uma interação espécie-específica com um organismo indutor. Os indutores manipulam o metabolismo do hospedeiro. Microgramma vacciniifolia (Langsd. & Fisch.) Copel. é uma samambaia epífita neotropical que hospeda duas galhas caulinares, uma induzida por uma espécie de mosquito (Diptera) e outra por uma micromariposa (Lepidoptera). O objetivo deste estudo foi avaliar o impacto desses dois insetos indutores de galhas na bioquímica dos ácidos fenólicos e da cianogênese em galhas, caules e folhas de M. vacciniifolia. Em análise de cromatografia líquida de alta eficiência (CLAE) foi possível indicar a presença de um total de 14 derivados fenólicos, incluindo ácido cafeico e ácido cumárico. Análise das Coordenadas Principais (ACoP) indicou três grupos (1) caules não galhados e galhas induzidas pela micromariposa; (2) galhas induzidas pelo mosquito; (3) galhas induzidas pelo mosquito com parasitoides. Em relação a frequência da cianogênese analisada com o teste do papel picrato, o teste do qui-quadrado apresentou diferença significativa entre as folhas férteis (8,3%), folhas estéreis (27,7%), caules não galhados (0%) e galhas. Entre as galhas, somente aquelas induzidas pelo mosquito foram cianogênicas (15%). Os resultados encontrados indicam, ao menos para as substâncias fenólicas, que os insetos indutores de galha (mosquito e micromariposa) promovem alterações espécie-específica na composição química da samambaia hospedeira.

Phenolic substances and cyanogenesis in galled and non-galled tissue of the fern species Microgramma vacciniifolia / Substâncias fenólicas e cianogênese em tecidos galhados e não galhados da samambaia Microgramma vacciniifolia

Galls, neo-formed plant structures that can occur in different organs, are generated by species-specific interaction with an inducing organism. Inducers manipulate the metabolism of its host. Microgramma vacciniifolia (Langsd. & Fisch.) Copel. is a Neotropical epiphytic fern that hosted two stem galls, one induced by a midge species (Diptera) and other by a micromoth species (Lepidoptera). The aim of this study was to evaluate the impact of these two gall-inducing insects on the biochemistry of phenolic acids and the cyanogenesis in galls, stems and leaves of M. vacciniifolia. High performance liquid chromatography (HPLC) indicated a total of 14 phenol derivatives, including caffeic and coumaric acid. Principal Coordinates Analysis (PCoA) of the phenolic substances indicated three groups consisting (1) non-galled stems and micromoth-induced galls; (2) midge-induced galls; (3) midge induced galls with parasitoids. Regarding the frequency of cyanogenesis assessed by the picrate paper test, the chi-squared test showed significant difference between fertile leaves (8.3%), sterile leaves (27.7%), non-galled stems (0%) and galls. Among galls, only the midge-induced galls analyzed were cyanogenic (15%). Our results indicated that the different gall-inducers (midge and micromoth) promote species-specific alterations to the phenolic substance composition of the host fern.

Galhas são estruturas vegetais neo-formadas que ocorrem em diferentes órgãos. Elas são geradas por uma interação espécie-específica com um organismo indutor. Os indutores manipulam o metabolismo do hospedeiro. Microgramma vacciniifolia (Langsd. & Fisch.) Copel. é uma samambaia epífita neotropical que hospeda duas galhas caulinares, uma induzida por uma espécie de mosquito (Diptera) e outra por uma micromariposa (Lepidoptera). O objetivo deste estudo foi avaliar o impacto desses dois insetos indutores de galhas na bioquímica dos ácidos fenólicos e da cianogênese em galhas, caules e folhas de M. vacciniifolia. Em análise de cromatografia líquida de alta eficiência (CLAE) foi possível indicar a presença de um total de 14 derivados fenólicos, incluindo ácido cafeico e ácido cumárico. Análise das Coordenadas Principais (ACoP) indicou três grupos (1) caules não galhados e galhas induzidas pela micromariposa; (2) galhas induzidas pelo mosquito; (3) galhas induzidas pelo mosquito com parasitoides. Em relação a frequência da cianogênese analisada com o teste do papel picrato, o teste do qui-quadrado apresentou diferença significativa entre as folhas férteis (8,3%), folhas estéreis (27,7%), caules não galhados (0%) e galhas. Entre as galhas, somente aquelas induzidas pelo mosquito foram cianogênicas (15%). Os resultados encontrados indicam, ao menos para as substâncias fenólicas, que os insetos indutores de galha (mosquito e micromariposa) promovem alterações espécie-específica na composição química da samambaia hospedeira.

Phenolic substances and cyanogenesis in galled and non-galled tissue of the fern species Microgramma vacciniifolia

Abstract Galls, neo-formed plant structures that can occur in different organs, are generated by species-specific interaction with an inducing organism. Inducers manipulate the metabolism of its host. Microgramma vacciniifolia (Langsd. & Fisch.) Copel. is a Neotropical epiphytic fern that hosted two stem galls, one induced by a midge species (Diptera) and other by a micromoth species (Lepidoptera). The aim of this study was to evaluate the impact of these two gall-inducing insects on the biochemistry of phenolic acids and the cyanogenesis in galls, stems and leaves of M. vacciniifolia. High performance liquid chromatography (HPLC) indicated a total of 14 phenol derivatives, including caffeic and coumaric acid. Principal Coordinates Analysis (PCoA) of the phenolic substances indicated three groups consisting (1) non-galled stems and micromoth-induced galls; (2) midge-induced galls; (3) midge-induced galls with parasitoids. Regarding the frequency of cyanogenesis assessed by the picrate paper test, the chi-squared test showed significant difference between fertile leaves (8.3%), sterile leaves (27.7%), non-galled stems (0%) and galls. Among galls, only the midge-induced galls analyzed were cyanogenic (15%). Our results indicated that the different gall-inducers (midge and micromoth) promote species-specific alterations to the phenolic substance composition of the host fern.

Resumo Galhas são estruturas vegetais neo-formadas que ocorrem em diferentes órgãos. Elas são geradas por uma interação espécie-específica com um organismo indutor. Os indutores manipulam o metabolismo do hospedeiro. Microgramma vacciniifolia (Langsd. & Fisch.) Copel. é uma samambaia epífita neotropical que hospeda duas galhas caulinares, uma induzida por uma espécie de mosquito (Diptera) e outra por uma micromariposa (Lepidoptera). O objetivo deste estudo foi avaliar o impacto desses dois insetos indutores de galhas na bioquímica dos ácidos fenólicos e da cianogênese em galhas, caules e folhas de M. vacciniifolia. Em análise de cromatografia líquida de alta eficiência (CLAE) foi possível indicar a presença de um total de 14 derivados fenólicos, incluindo ácido cafeico e ácido cumárico. Análise das Coordenadas Principais (ACoP) indicou três grupos (1) caules não galhados e galhas induzidas pela micromariposa; (2) galhas induzidas pelo mosquito; (3) galhas induzidas pelo mosquito com parasitoides. Em relação a frequência da cianogênese analisada com o teste do papel picrato, o teste do qui-quadrado apresentou diferença significativa entre as folhas férteis (8,3%), folhas estéreis (27,7%), caules não galhados (0%) e galhas. Entre as galhas, somente aquelas induzidas pelo mosquito foram cianogênicas (15%). Os resultados encontrados indicam, ao menos para as substâncias fenólicas, que os insetos indutores de galha (mosquito e micromariposa) promovem alterações espécie-específica na composição química da samambaia hospedeira.

Pest categorisation of Resseliella citrifrugis.

The EFSA Panel on Plant Health performed a pest categorisation of the citrus fruit midge, Resseliella citrifrugis Jiang (Diptera: Cecidomyiidae), for the territory of the EU. This species is not included in EU Commission Implementing Regulation 2019/2072. This oligophagous species, which feeds on Citrus spp. fruits, is known to occur in China (provinces of Fujian, Gansu, Guangdong, Guangxi, Guizhou, Hubei, Hunan, Jiangxi and Sichuan). First-generation adults emerge in April and oviposit on the peduncle and calyx of citrus fruits. Larvae feed on the albedo (the white layer between skin and pulp) of the fruit, which they tunnel. Infested fruit most often drop prematurely. Larvae pupate either within the fallen fruit or in the soil. This species has three main peaks of activity (May, June-July, August-September) and four generations per year in its native range. Mature larvae from the last generation are the overwintering stage. Potential entry pathways for R. citrifrugis, such as Citrus spp. plants for planting with foliage and soil/growing media, and soil/growing media can be considered as closed. The citrus fruit pathway remains open from countries where R. citrifrugis is known to occur. Indeed, this species was intercepted in fresh pomelos 11 times from December 2020 to January 2021 in the EU. Should R. citrifrugis enter the EU, the ample availability of hosts (Citrus spp.) and the climatic conditions in citrus-growing areas of southern EU Member States would most probably allow this species to successfully establish and spread. Economic impact in citrus production is anticipated if establishment occurs. R. citrifrugis satisfies the criteria that are within the remit of EFSA to assess for this species to be regarded as a potential Union quarantine pest. There is uncertainty about the nomenclature of R. citrifrugis and its exact host range. However, because the name Resseliella citrifrugis is used in multiple sources reporting it as a pest of citrus in China, where symptoms, biology and impact are described, these uncertainties do not affect the conclusions of this categorisation.