Gall Induction Research Articles

Lasioptera rubi, a Pest of Rubus idaeus: Galls Morphology, Anatomy and Histochemistry

Rubus species are affected by the gall inducer Lasioptera rubi (Diptera, Cecidomyiidae), which produces galls predominantly on the stem but sometimes also on the petiole. The presence of these neo-structures at the stem level can affect their survival and implicit production in raspberry crops. The purpose of the present study is to achieve the first detailed description of the histological and morphological peculiarities of these galls, which present great variability from these points of view. Three morphotypes were described in the galls produced in the stem, depending on the position of the larval chambers and the degree of damage to the vascular system. The histochemical investigations highlighted the presence of reducing sugars and proteins as the main nutrients in the gall tissues, as well as the presence of polyphenols and terpenoids as secondary products of metabolism. Indoleacetic acid was identified near the larval chambers and in the neighboring tissues, its presence having an important role in the changes in the morphogenesis of the secondary xylem in the stem and in inducing the appearance of meristematic nodules in the gall structure.

How does the life cycle of Clinodiplosis profusa (Cecidomyiidae) adjust to phenological variations of the host plant Eugenia uniflora (Myrtaceae) in sun and shade?

Galls are plant neoformations induced by specialized parasites. Since gall inducers rely on reactive plant sites for gall development, variations in abiotic factors that affect plant phenology are expected to impact the life cycle of gall inducers. To test the hypothesis that different light conditions affect both host plant and gall inducer life cycles, we studied the system Eugenia uniflora (Myrtaceae) - Clinodiplosis profusa (Cecidomyiidae), comparing plants occurring in sunny and shaded environments. We mapped phenological differences among individuals of E. uniflora occurring in the two environments and related them to the influence of luminosity on the life cycle of the gall inducer. Shade plants showed lower intensity of leaf sprouting throughout the year compared to sun-exposed plants, especially during the rainy season. Young and mature galls are synchronized with the peak of leaf sprouting at the beginning of the rainy season, lasting longer in sun-exposed plants - approximately two months longer compared to shade plants. The greater light intensity positively impacts the formation and growth of leaves and galls, with an extended period available for their induction and growth. Thus, light is an important factor for the development of gallers, considering that variations in luminosity influenced not only the phenology of the host plant, but also determined the life cycle of gall inducers. Furthermore, changes in plant-environment interactions are expected to affect the life cycle and richness of other host plant-gall inducer systems.

Ontogenetic differences in sun and shade galls of Clinodiplosis profusa on Eugenia uniflora leaves and the cytological antioxidant mechanisms in gall cells.

The gall-host Eugenia uniflora (Myrtaceae) is adaptable to different light conditions, enabling leaf production and survival in both sun and shade. Leaves of E. uniflora in shaded environments have more mesophyll layers, and galls of Clinodiplosis profusa (Cecidomyiidae) are larger and wider. Based on these previous observations, this study investigated the morphogenesis of galls induced by C. profusa on leaves of E. uniflora in different light conditions, revealing if the galls have a potential for acclimation, as observed with leaves. For this purpose, we compared the anatomical, histometric, and histochemical development of leaves and galls at different stages of development in sun and shade environments. Additionally, we analyzed the cytological features of the tissues composing the mature gall walls. Cells of shade galls expanded more toward the end of the developmental phase, which may explain the larger volume found for shade galls in a previous study. However, during the mature phase, these galls showed no significant differences in tissue thickness and final cell elongation in the contrasting light conditions. In the ultrastructural analyses, mature galls showed a gradient distinguishing the outer and inner parenchyma cells. The inner parenchyma had nutritive cells, with dense cytoplasm and abundant organelles. A higher accumulation of starch grains in nutritive cells, with evidence of hydrolysis of starch grains detected in the innermost layers leads to the accumulation of reducing sugars, which, with the presence of plastoglobules and protein bodies, are important mechanisms of oxidative stress dissipation in the cells in contact with the gall inducer.

Successful gall induction on wild roses by gall wasps under lab conditions

Successful gall induction on wild roses by gall wasps under lab conditions

New species of gall wasps (Hymenoptera: Cynipidae: Aulacideini and Phanacidini) inducing galls on plants of the genera Sonchus and Crepis (Asteraceae) in Romania (Dobrogea Province) and notes on their parasitoids

Two new species of herb gall wasps (Hym., Cynipidae) developing on plants in the genera Sonchus L. and Crepis L. (Asteraceae) are described from Southeastern Romania (Dobrogea Province): Aulacidea andriescui sp. n. gall inducer on Sonchus palustris L. and Phanacis dobrogicus sp. n. gall maker on Crepis foetida subsp. rhoeadifolia (M. Bieb.) Čelak. In addition to the diagnosis, identification, morphological description, life-cycle, geographical distribution, photos of gall and host plant, and SEM images, data on the associated fauna obtained from the galls of the two new species are also presented.

Community structure of parasitoids attacking Schizomyia sasakii (Diptera: Cecidomyiidae) and local adaptation of the gall midge in the Izu region

AbstractInsect community structures and biological interactions vary with the distance from the mainland to islands. Gall inducers are key organisms in local arthropod communities because their galls harbor diverse arthropods. We investigated the parasitoid community of a gall‐inducing cecidomyiid Schizomyia sasakii on the Izu Peninsula and the Izu Islands, Japan. We examined relationships between parasitism by Inostemma sp. and Torymus hirtipennis, and gall characteristics, and analyzed the directional selection on gall characteristics induced by S. sasakii in each locality. The species richness of parasitoids on the Izu Islands decreased with the distance from the Izu Peninsula to each island, and area of respective islands. Inostemma sp. preferably attacked large galls on Ohshima Island, and T. hirtipennis tended to attack relatively small galls in the Izu Peninsula. Directional selection on gall characteristics of S. sasakii favored the induction of larger galls and thicker tissues by S. sasakii on Ohshima Island. In contrast, no directional selection was detected in the gall characteristics on Hachijojima Island. The number of alternative host species of parasitoid may affect variation in parasitoid richness of S. sasakii on distant and small islands, because the species richness of gall midges is necessary to maintain local parasitoid populations. Our study supported the ovipositor limitation hypothesis for the torymid species, related to gall size. The directional selection of parasitoid attacks on the Izu Islands may act to favor the induction of large and hypertrophic galls by S. sasakii to avoid the parasitoids.

Implications of cell wall immunocytochemical profiles on the structural and functional traits of root and stem galls induced by Eriosoma lanigerum on Malus domestica.

Alterations in cell wall composition imply in new structural and functional traits in gall developmental sites, even when the inducer is a sucking exophytophagous insect with strict feeding sites as the aphid associated to Malus domestica Borkh. This host plant is an economically important, fruit-bearing species, susceptible to gall induction by the sucking aphid Eriosoma lanigerum Hausmann, 1802. Herein, the immunocytochemical detection of arabinogalactan-proteins (AGPs), pectins, and hemicelluloses using monoclonal antibodies was performed in samples of non-galled roots and stems, and of root and stem galls on M. domestica. The dynamics of these cell wall components was discussed under the structural and functional traits of the galls proximal, median, and distal regions, according to the proximity of E. lanigerum colony feeding site. In the proximal region, the epitopes of AGPs and homogalacturonans (HGs) are related to cell growth and divisions, which result in the overproduction of parenchyma cells both in root and stem galls. In the proximal and median regions, the co-occurrence of HGs and arabinans in the cell walls of parenchyma and secondary tissues favors the nutrient flow and water-holding capacity, while the xylogalacturonans and hemicelluloses may function as additional carbohydrate resources to E. lanigerum. The immunocytochemical profile of the cell walls support the feeding activity of E. lanigerum mainly in the gall proximal region. The similarity of the cell wall components of the gall distal region and the non-galled portions, both in roots and stems, relates to the decrease of the cecidogenetic field the more distant the E. lanigerum colony is.

Gall-inducing and building habit in Lixinae weevils (Coleoptera: Curculionidae)

Among the Lixinae, at least 35 species are gall inducers. Twenty six species have larvae with building activity (lists of both are provided). Fifteen species are root gallers (tribe Cleonini) and 20 species are stem gallers (Lixini). Their gall morphology (shape, colour, size), habitat peculiarities and typical gall development are described. A new classification of galls according to their inhabitants and formation is proposed. The data obtained suggest that galling depends on the thickness of the transformed plant parts. The potential benefits and costs of galling to gall inducers are discussed. Data on the diversity of parasitoids indirectly support a protective role for galls. A summary of information on special larval constructions is presented. Two types of such constructions are—the ground capsules and the trehalas. These are closed chambers whose walls consist of a small soil and plant particles bound with sticky larval secretions. Capsules and trehalas are attached to the plant and serve to complete preimaginal development. Both are functionally similar to galls.

Gall inducer Dasineura sp. alters the polyphenol profile and antioxidant activity of Peumus boldus stems.

Peumus boldus, a tree native to Chile, is extensively used for medicinal purposes due to its richness in alkaloids and antioxidant polyphenols. A species of galling insect, Dasineura sp. induces structural and chemical changes on P. boldus stems while its galls are established and developed. Taking into account the antioxidant properties of P. boldus polyphenols, it would be expected that Dasineura sp. induces changes in the accumulation sites, chemical profile, and antioxidant activity of the P. boldus stem polyphenols, related to different reactive oxygen species (ROS) production levels during gall development. Dasineura sp. induces changes in the accumulation sites of total polyphenols, flavonols, and lignin, redirecting their accumulation toward the sites of greatest production of H2O2 and O2.-. Although changes in total polyphenol content would be expected, this did not vary significantly between non-galled and galled stems. However, the galling insect induced changes in the profile and concentration of soluble polyphenols, leading to the gall extracts' antioxidant capacity decreasing significantly during the maturation and senescence stages. Additionally, during the maturation stage, lignin deposition increases in the more peripheral gall tissues, which also contributes to ROS dissipation. The differences in the different gall developmental stages' antioxidant activity could be related to the identity and concentration of phenolic compounds in each gall extract, rather than to the total phenol content. Regardless of the mechanisms involved, the dissipation of the ROS generated by Dasineura sp. activity occurs, restoring the redox balance in galls and guaranteeing the success of the inducer.

Cynipid wasps systematically reprogram host metabolism and restructure cell walls in developing galls.

Many insects have evolved the ability to manipulate plant growth to generate extraordinary structures called galls, in which insect larva can develop while being sheltered and feeding on the plant. In particular, cynipid (Hymenoptera: Cynipidae) wasps have evolved to form morphologically complex galls and generate an astonishing array of gall shapes, colors, and sizes. However, the biochemical basis underlying these remarkable cellular and developmental transformations remains poorly understood. A key determinant in plant cellular development is cell wall deposition that dictates the physical form and physiological function of newly developing cells, tissues, and organs. However, it is unclear to what degree cell walls are restructured to initiate and support the formation of new gall tissue. Here, we characterize the molecular alterations underlying gall development using a combination of metabolomic, histological, and biochemical techniques to elucidate how valley oak (Quercus lobata) leaf cells are reprogrammed to form galls. Strikingly, gall development involves an exceptionally coordinated spatial deposition of lignin and xylan to form de novo gall vasculature. Our results highlight how cynipid wasps can radically change the metabolite profile and restructure the cell wall to enable the formation of galls, providing insights into the mechanism of gall induction and the extent to which plants can be entirely reprogrammed to form unique structures and organs.

New Gall-Forming Insect Model, Smicronyx madaranus: Critical Stages for Gall Formation, Phylogeny, and Effectiveness of Gene Functional Analysis.

The molecular mechanisms underlying insect gall formation remain unclear. A major reason for the inability to identify the responsible genes is that only a few systems can be experimentally validated in the laboratory. To overcome these problems, we established a new galling insect model, Smicronyx madaranus. Our manipulation experiments using nail polish sealing and insecticide treatment revealed an age-dependent change in gall formation by S. madaranus; adult females and larvae are responsible for gall induction and enlargement, respectively. Furthermore, it has been suggested that substances released during oviposition and larval feeding are involved in each process. Phylogenetic analysis showed that gall-forming weevils, including S. madaranus, belong to two distinct lineages that utilize different host plants. This may indicate that gall-forming traits evolved independently in these Smicronyx lineages. The efficacy of RNA interference (RNAi) in S. madaranus was confirmed by targeting the multicopper oxidase 2 gene. It is expected that the mechanisms of gall formation will be elucidated by a comprehensive functional analysis of candidate genes using RNAi and the S. madaranus galling system in the near future.

Cell wall epitope distribution in the functional compartments of galls induced by Palaeomystella oligophaga (Lepidoptera) in Macairea radula (Melastomataceae)

Context The parasitic interaction between the galling insect Palaeomystella oligophaga (Lepidoptera) and the host plant tissues of Macairea radula (Melastomataceae) leads to the formation of globoid galls. These galls have storage and typical nutritive tissues (outer and inner compartments, respectively), whose functions may be related to differential cell wall component distributions and to the stage of development of the gall inducer. Aims We evaluated whether the cell wall composition of the gall compartments (storage and nutritive tissues) changes according to the developmental stage of the gall inducer P. oligophaga. Methods The galls occupied by the gall inducer in different stages of development (i.e. larval and pupal stages, as well as empty galls) were collected and submitted to histological examination, immunocytochemical analyses using monoclonal antibodies for pectins, hemicelluloses, and glycoproteins, and histochemical analyses using phloroglucinol for lignins. Key results The histological results showed that nutritive tissues have smaller cells compared with storage ones when the galling insect is in the larval stage. Immunocytochemical analyses also showed the occurrence of both methyl-esterified and unesterified pectins, as well hemicelluloses in the nutritive tissue during the larval stage. The dynamic of some cell wall components changes when the gall inducer is in the pupal stage, or the galls are empty. Conclusions The changes in the composition of the cell walls, especially regarding homogalacturonans and xyloglucans, support the high metabolism and resource availability in cell walls of nutritive tissue for the gall inducer. Implications The cell wall composition may indicate important metabolic steps during gall formation and galling insect diet.

Evergreen forests on the Korean Peninsula faced a new threat: Schizomyia castanopsisae (Jat-bam-na-mu-hok-pa-ri) inducing galls on reproductive organs of Castanopsis sieboldii in the Yokjido Island Natural Monument Forest

Unnoticed and silently, a certain midge has emerged that could pose a threat to the evergreen forests on the Korean Peninsula and the associated island ecosystems. We report the discovery of an unrecorded species on the Korean Peninsula, identified as Schizomyia castanopsisae Elsayed & Tokuda, 2018 (Diptera: Cecidomyiidae). This previously unrecorded gall midge species induces galls on a reproductive organ of Castanopsis sieboldii, specifically the inflorescences, and was found in the natural monument forest on Yokjido Island. A total of 40 larvae were collected from 8 inflorescence galls, each obtained from individual hosts within the forest. Morphological examinations and mitochondrial DNA cytochrome oxidase subunit I (COI) region sequencing were conducted for each specimen, confirming the identification of the gall inducer as S. castanopsisae. The COI sequences from the 40 specimens collected on Yokjido Island were identical, showing a 99.8% match with individuals from Kyushu, Japan. Additionally, a maximum likelihood tree analysis revealed a sister relationship between Yokjido Island and Kyushu individuals. Considering the tropical nature of gall midge species, it suggests recent northward migration from Kyushu to Yokjido Island. This species not only significantly reduces acorn production but also has the potential to negatively impact organisms associated with acorns. Drawing from cases in Japan, this species has the potential to rapidly dominate evergreen broad-leaved forests on numerous islands in the Korean Peninsula. Therefore, urgent additional research on aspects such as natural enemies of this species, its dispersal mechanisms, and other relevant factors is necessary. We also investigated the distribution of trees with inflorescences gall-induced by S. castanopsisae in the Yokjido Island Natural Monument Forest. The occurrence frequency of these galls is estimated to be approximately 10 percent.

The regeneration conferring transcription factor complex ERF115-PAT1 coordinates a wound-induced response in root-knot nematode induced galls.

The establishment of root-knot nematode (RKN; Meloidogyne spp.) induced galls in the plant host roots likely involves a wound-induced regeneration response. Confocal imaging demonstrates physical stress or injury caused by RKN infection during parasitism in the model host Arabidopsis thaliana. The ERF115-PAT1 heterodimeric transcription factor complex plays a recognized role in wound-induced regeneration. ERF115 and PAT1 expression flanks injured gall cells likely driving mechanisms of wound healing, implying a local reactivation of cell division which is also hypothetically involved in gall genesis. Herein, functional investigation revealed that ectopic ERF115 expression resulted in premature induction of galls, and callus formation adjacent to the expanding female RKN was seen upon PAT1 upregulation. Smaller galls and less reproduction were observed in ERF115 and PAT1 knockouts. Investigation of components in the ERF115 network upon overexpression and knockdown by qRT-PCR suggests it contributes to steer gall wound-sensing and subsequent competence for tissue regeneration. High expression of CYCD6;1 was detected in galls, and WIND1 overexpression resulted in similar ERF115OE gall phenotypes, also showing faster gall induction. Along these lines, we show that the ERF115-PAT1 complex likely coordinates stress signalling with tissue healing, keeping the gall functional until maturation and nematode reproduction.

Microbiome and plant cell transformation trigger insect gall induction in cassava.

Several specialised insects can manipulate normal plant development to induce a highly organised structure known as a gall, which represents one of the most complex interactions between insects and plants. Thus far, the mechanism for insect-induced plant galls has remained elusive. To study the induction mechanism of insect galls, we selected the gall induced by Iatrophobia brasiliensis (Diptera: Cecidomyiidae) in cassava (Euphorbiaceae: Manihot esculenta Crantz) as our model. PCR-based molecular markers and deep metagenomic sequencing data were employed to analyse the gall microbiome and to test the hypothesis that gall cells are genetically transformed by insect vectored bacteria. A shotgun sequencing discrimination approach was implemented to selectively discriminate between foreign DNA and the reference host plant genome. Several known candidate insertion sequences were identified, the most significant being DNA sequences found in bacterial genes related to the transcription regulatory factor CadR, cadmium-transporting ATPase encoded by the cadA gene, nitrate transport permease protein (nrtB gene), and arsenical pump ATPase (arsA gene). In addition, a DNA fragment associated with ubiquitin-like gene E2 was identified as a potential accessory genetic element involved in gall induction mechanism. Furthermore, our results suggest that the increased quality and rapid development of gall tissue are mostly driven by microbiome enrichment and the acquisition of critical endophytes. An initial gall-like structure was experimentally obtained in M. esculenta cultured tissues through inoculation assays using a Rhodococcus bacterial strain that originated from the inducing insect, which we related to the gall induction process. We provide evidence that the modification of the endophytic microbiome and the genetic transformation of plant cells in M. esculenta are two essential requirements for insect-induced gall formation. Based on these findings and having observed the same potential DNA marker in galls from other plant species (ubiquitin-like gene E2), we speculate that bacterially mediated genetic transformation of plant cells may represent a more widespread gall induction mechanism found in nature.

Phylogenomic analysis of protein‐coding genes resolves complex gall wasp relationships

AbstractGall wasps (Hymenoptera: Cynipidae) comprise 13 distinct tribes whose interrelationships remain incompletely understood. Recent analyses of ultra‐conserved elements (UCEs) represent the first attempt at resolving these relationships using phylogenomics. Here, we present the first analysis based on protein‐coding sequences from genome and transcriptome assemblies. Unlike UCEs, these data allow more sophisticated substitution models, which can potentially resolve issues with long‐branch attraction. We include data for 37 cynipoid species, including two tribes missing in the UCE analysis: Aylacini (s. str.) and Qwaqwaiini. Our results confirm the UCE result that Cynipidae are not monophyletic. Specifically, the Paraulacini and Diplolepidini + Pediaspidini fall outside a core clade (Cynipidae s. str.), which is more closely related to the insect‐parasitic Figitidae, and this result is robust to the exclusion of long‐branch taxa that could mislead the analysis. Given this, we here divide the Cynipidae into three families: the Paraulacidae stat. prom., Diplolepididae stat. prom. and Cynipidae (s. str.). Our results suggest that the Eschatocerini are the sister group of the remaining Cynipidae (s. str.). Within the Cynipidae (s. str.), the Aylacini (s. str.) are more closely related to oak gall wasps (Cynipini) and some of their inquilines (Ceroptresini) than to other herb gallers (Aulacideini and Phanacidini), and the Qwaqwaiini likely form a clade together with Synergini (s. str.) and Rhoophilini. Several alternative scenarios for the evolution of cynipid life histories are compatible with the relationships suggested by our analysis, but all are complex and require multiple shifts among parasitoids, inquilines and gall inducers.

Epidermal and subepidermal changes during the formation of hairy galls induced by Eriophyidae on Avicennia schaueriana leaves.

Leaf-galling Eriophyidae (Acarina) may promote simple or complex alterations in the organs of their host plants, such as an increase in indumentum density or the reorganization of epidermis and ground system tissue patterns. To test if hairy galls of Eriophyidae on Avicennia schaueriana (Acanthaceae) are related to complex changes, leaf galls in distinct developmental phases were compared to non-galled leaves using anatomical, histochemical, and histometric analyses. Quantitative comparisons of preferential gall induction sites and gall area according to distinct leaf portions were made to evaluate if the impacts of gall formation can be related to the distinct potentialities of leaf microsites. The apical portion of the leaves and leaf margins were the sites with the highest occurrence of galls, but no relationship was detected between gall area and induction site. The gall anatomy revealed that epidermal features are influenced the most with the development of abnormal stomata and projected or sunken salt glands. The most striking change is the neoformation of elongated filiform trichomes on the abaxial surface (where the mites occur) that accumulate reducing sugars and proteins. The filiform trichomes may protect the inducers against abiotic stressors and enemies, and the primary metabolites that accumulate are important foods for mites. The mesophyll has simple alterations, only in the spongy parenchyma. Complex alterations occur only in abaxial epidermal cells close to feeding sites of the inducer. The number of inducers per gall seems to be the most important influence on gall size, since gall area is not related to the position in the leaves.

Comparative anatomy of venom glands suggests a role of maternal secretions in gall induction by cynipid wasps (Hymenoptera: Cynipidae)

Abstract Many herbivorous insect species are capable of hijacking plant development to induce novel plant organs called galls. In most groups of galling insects, the insect organs and molecular signals involved in gall induction are poorly understood. We focused on gall wasps (Hymenoptera:Cynipidae), the second largest clade of gall inducers (~1,400 spp.), for which the developmental stages and organs responsible for gall development are unclear. We investigated the female metasomal anatomy of 69 gall-inducing and 29 non-gall-inducing species across each of the major lineages of Cynipoidea, to test relationships between this lifestyle and the relative size of secretory organs. We confirmed that the venom apparatus in gall-inducing species is greatly expanded, although gall-inducing lineages vary in the relative size of these glands. Among these gallers, we measured the largest venom gland apparatus relative to body size ever recorded in insects. Non-galling inquiline species are accompanied by a reduction of this apparatus. Comparative microscopic analysis of venom glands suggests varying venom gland content across the lineages. Some oak gallers also had enlarged accessory glands, a lipid-rich organ whose function remains unclear, and which has not been previously studied in relation to gall formation. Together, the massive expansion of secretory organs specifically in gall-inducing species suggests a role of these secretions in the process of gall formation, and the variance in size of venom glands, accessory glands, and the contents of these glands among gallers, suggests that gall formation across this clade is likely to employ a diversity of molecular strategies.

Biology and morphometric relationship of gall inducers Contarinia sp. and corresponding parasitoids for swollen galls of Nitraria sibirica pall.

Galls function as provide shelter for gall inducers, guarding them against their natural enemies. Previous research has illuminated the interactions between galls, gall inducers, and their corresponding parasitoids within various caltrop plants. However, less is known about these relationships within Nitraria sibirica, particularly regarding the efficacy of parasitism. Therefore, this study aimed to identify the morphometric relationships among the swollen galls, gall inducers, and their parasitoids. Two species of gall inducers and three species of parasitoids were obtained from the swollen galls of N. sibirica. The correlations of the parasitization indexes, the lifespan of gall inhabitants, and temperature and the morphometric relationships between the galls and their inhabitants were analyzed. The dominant gall inducer identified was Contarinia sp. (Diptera: Cecidomyiidae). Furthermore, it was observed that three solitary parasitoids attacked Contarinia sp. in the swollen galls, with only Eupelmus gelechiphagus acting as an idiobiont ectoparasitoid. The dominant parasitoids were Platygaster sp. and Cheiloneurus elegans at sites 1 and 2, respectively, with Platygaster sp. displaying greater abundance than C. elegans in the swollen galls. The lifespan of the gall inhabitants shortened gradually as the temperature increased. Moreover, the optimal number of gall chambers ranged from two to four per swollen gall with maximized fitness, which can be considered the optimal population density for the gall inducer Contarinia sp. Morphometric analysis exhibited a strong linear correlation between gall size and chamber number or the number of gall inhabitants, as well as a weak correlation between gall size and body size of the primary inhabitants of swollen galls. Our results highlight the importance of the biological investigation of parasitoids and gall inducers living in closed galls with multiple chambers and may pave the way for potential application in biological control.

Conferring High IAA Productivity on Low-IAA-Producing Organisms with PonAAS2, an Aromatic Aldehyde Synthase of a Galling Sawfly, and Identification of Its Inhibitor.

Gall-inducing insects often contain high concentrations of phytohormones, such as auxin and cytokinin, which are suggested to be involved in gall induction, but no conclusive evidence has yet been obtained. There are two possible approaches to investigating the importance of phytohormones in gall induction: demonstrating either that high phytohormone productivity can induce gall-inducing ability in non-gall-inducing insects or that the gall-inducing ability is inhibited when phytohormone productivity in galling insects is suppressed. In this study, we show that the overexpression of PonAAS2, which encodes an aromatic aldehyde synthase (AAS) responsible for the rate-limiting step in indoleacetic acid (IAA) biosynthesis in a galling sawfly (Pontania sp.) that contains high levels of endogenous IAA, conferred high IAA productivity on Caenorhabditis elegans, as the model system. This result strongly suggests that PonAAS2 can also confer high IAA productivity on low-IAA-producing insects. We also successfully identified an inhibitor of PonAAS2 in a chemical library. This highly selective inhibitor showed stronger inhibitory activity against AAS than against aromatic amino acid decarboxylase, which belongs to the same superfamily as AAS. We also confirm that this inhibitor clearly inhibited IAA productivity in the high-IAA-producing C. elegans engineered here.