Parasitoid impairment on the galling Lopesia sp. activity reflects on the cytological and histochemical profiles of the globoid bivalve-shaped gall on Mimosa gemmulata.

Gall cytological and histochemical features established by the constant feeding activity of the associated gall inducer may be changed due to the attack of parasitoids. We accessed two tri-trophic systems involving the globoid bivalve-shaped gall on Mimosa gemmulata Barneby (Fabaceae) and its galling undescribed species of Lopesia (Diptera: Cecidomyiidae), which may be ectoparasitized by Torymus sp. (Hymenoptera: Torymidae) or endoparasitized by a polyembryonic Platygastridae (Hymenoptera), as models of study. The ectoparasitoid species paralyzes and kills Lopesia sp. larva, which stops the feeding stimuli, while the endoparasitoid larvae feed in Lopesia sp. larva body and keep it alive for a certain time. Our hypothesis is that the time lapse of Lopesia sp. feeding impairment by the two parasitoids will cause distinct cytological and histochemical responses in the ecto- and endoparasitized galls compared to the non-parasitized condition. In both parasitoidism cases, the impairment of the feeding activity of the galling Lopesia sp. directs the common storage and nutritive cells toward a similar process of induced cell death, involving cell collapse and loss of membrane integrity. The cell metabolism is maintained mainly by mitochondria, and by the translocation of lipids from the common storage tissue, via plasmodesmata, through the living sclereids of the mechanical zone toward the nutritive tissue. Accordingly, the parasitoid impairment on the feeding activity of Lopesia sp. larvae causes precocious senescence, but similar cytological alterations, and no impact over the histochemical profiles, regarding lipids, reactive oxygen species, and secondary metabolites, which support gall metabolism along the parasitoid cycles.

Structural and Nutritional Peculiarities Related to Lifespan Differences on Four Lopesia Induced Bivalve-Shaped Galls on the Single Super-Host Mimosa gemmulata.

Super-host plants are elegant models to evaluate the peculiarities of gall structural and nutritional profiles due to the stimuli of distinct gall inducers in temporal and spatial perspectives. Galls induced by congeneric insects, Lopesia spp. (Diptera, Cecidomyiidae) on the same host plant, Mimosa gemmulata Barneby (Fabaceae) were analyzed to estimate if variations of 1 or 2 months in gall lifespans may result in differences over the accumulation of nutritional resources, and their compartmentalization both in cell walls and protoplasm. Mimosa gemmulata hosts four Lopesia-induced galls: the lenticular bivalve-shaped gall (LG) with a 2-month life cycle, the brown lanceolate bivalve-shaped gall (BLG) and the green lanceolate bivalve-shaped gall (GLG) with 3 month-life cycles, and the globoid bivalve-shaped gall (GG) with a 4 month-life cycle. The comparisons among the four Lopesia galls, using anatomical, histometric, histochemical, and immunocytochemical tools, have demonstrated that the longest lifespan of the GG related to its highest increment in structural and nutritional traits compared with the LG, GLG, and BLG. The differences among the tissue stratification and cell wall thickness of the galls with the 2-month and the 3-month lifespans were subtle. However, the GG had thicker cell walls and higher stratification of the common storage tissue, schlerenchymatic layers and typical nutritive tissue than the other three gall morphospecies. The higher tissue thickness of the GG was followed by the formation of a bidirectional gradient of carbohydrates in the protoplasm, and the detection of xyloglucans in cell walls. Current data supported the presumption that the longest the lifespan, the highest the impact over the structural and nutritional metabolism of the Lopesia galls associated to M. gemmulata.

Pseudophacopteron longicaudatum (Hemiptera) induces intralaminar leaf galls on Aspidosperma tomentosum (Apocynaceae): a qualitative and quantitative structural overview.

The structural complexity of galls depends on species-specific interaction driven by the galling taxa. However, the host plant and environment stressors can impose limits on gall developmental patterns and impact the establishment of gall morphology. Herein, we employed qualitative and quantitative approaches in order to elucidate how cell divisions, elongation patterns, and tissue organization are determinant for the development of intralaminar gall morphology induced by Pseudophacopteron longicaudatum Malenovský, Burckhardt, Queiroz, Isaias & Oliveira (Hemiptera: Psylloidea: Phacopteronidae) on leaves of Aspidosperma tomentosum Mart. (Apocynaceae). In addition, we aimed to determine which anatomical process can discriminate the stages of gall development, plus, examine the histochemical and cytological profiles of the galls. The differentiated structures, mainly abaxial epidermis and spongy parenchyma, are associated with gall closure, with hyperplastic events concentrated in the young phase of the galls. Thus, epidermis and spongy parenchyma hypertrophy and are responsible for the determination of the nymphal chamber formation and gall shape. The mature galls do not differentiate into a typical nutritive cells and do not develop a histochemical gradient in their tissues. The cytological features of galls such as plastoglobules and multivesicular bodies are related to ROS scavenging mechanisms due the high oxidative stress.

Correction to: How the activity of natural enemies changes the structure and metabolism of the nutritive tissue in galls? Evidence from the Palaeomystella oligophaga (Lepidoptera) -Macairea radula (Metastomataceae) system.

The original version of this article unfortunately contains an error. The correct caption of Figures 2 and 3 are shown in this paper.

Red galls: the different stories of two gall types on the same host.

Several studies have suggested reasons why galls have conspicuous colours, but none of the ideas have been confirmed. However, what if the vibrant colours of some galls are explained simply by the effect of light exposure? This may lead to anthocyanin accumulation, functioning as a defence mechanism against the effects of high light. We studied the globoid galls induced by Cecidomyiidae (Diptera) on Qualea parviflora (Vochysiaceae), relating anthocyanin accumulation and chlorophyll fluorescence parameters to light incidence in abaxial and adaxial galls. We also tested if the anthocyanin accumulation patterns apply to another Cecidomyiidae-induced gall morphotype (intralaminar) within the same plant. Adaxial galls are exposed to higher incident light, with more anthocyanin accumulation and therefore red coloration. In galls from angled leaves, the greater the angle of the leaf, the higher the difference between anthocyanins on the sun and shade sides of galls. Photosynthetic pigment concentrations did not differ between abaxial and adaxial galls. However, we found higher (Fm ' - F')/Fm ' and Fv /Fm in the abaxial galls. Conversely, NPQ and Rfd were higher in adaxial galls. Finally, the pattern of anthocyanin accumulation was not found in the intralaminar gall. Anthocyanin accumulation in galls functions as a photoprotective strategy, maintaining tissue vitality in regions exposed to high light conditions. However, this mechanism may vary even among galls within the same host, indicating idiosyncrasy when it comes to coloration in galls. To date, this is the first study to demonstrate quantitatively why the galls of a specific species may be coloured: the variation in light regimes creates differential anthocyanin accumulation, influencing coloration.

How the activity of natural enemies changes the structure and metabolism of the nutritive tissue in galls? Evidence from the Palaeomystella oligophaga (Lepidoptera) -Macairea radula (Metastomataceae) system.

Insect-induced galls usually develop nutritional cells, which they induce and consume directly, and any metabolic modification of those cells may reflect changes of the insect's own metabolism. The system Palaeomystella oligophaga (Lepidoptera)-Macairea radula (Melastomataceae) presents a series of natural enemies, including parasitoids and cecidophages that can function as a natural experiment, respectively removing the specific galling feeding stimulus and providing a nonspecific one. Considering that the process of induction and maintenance of gall tissues strictly depends on the constant specific stimulus of galling, question I:What kind of metabolic changes these different groups of natural enemies can promote in chemical and structural composition of these galls? II: How the specialized tissues are metabolically dependent on the constant specific stimulus of galling in latter stages of gall development? Galls without natural enemies, with parasitoids or cecidophages in larvae or pupae stages were analyzed through histochemistry and cytological profiles and all compared to galls in natural senescence state. The analysis revealed the accumulation of proteins and lipids in typical nutritive tissue and starch in the storage tissue, as well a high integrity of cellular organelles and membrane systems on galls with gallings in the larval stage. Both parasitoids and cecidophages stop galling feeding activities, which resulted in the paralysis of the stimulus that maintain the metabolism of gall tissues, leading to generalized collapse. We demonstrate that the development and metabolic maintenance of a typical nutritive tissue in these galls are completely dependent on constant larval stimulus.

Antioxidant metabolism in galls due to the extended phenotypes of the associated organisms.

Animal-induced galls are considered extended phenotypes of their inducers, and therefore plant morphogenesis and metabolism may vary according to the species of gall inducers. The alterations in vacuolar and apoplastic polyphenols, carotenoids, chlorophyll fluorescence rates, PSII quantum yield, and phospholipid peroxidation were studied in galls induced by Ditylenchus gallaeformans (Nematoda) on Miconia albicans and M. ibaguensis (Melastomataceae), and by an unidentified Eriophyidae (Acarina) on M. ibaguensis. The focus currently addressed is gall metabolism as the extended phenotype of the gall inducers, and the neglected determination of gall functionalities over host plant peculiarities. Galls induced by D. gallaeformans on M. albicans and by the Eriophyidae on M. ibaguensis have increased accumulation of apoplastic and vacuolar phenolics, which is related to the control of phospholipid peroxidation and photoprotection. The galls induced by D. gallaeformans on M. ibaguensis have higher carotenoid and vacuolar polyphenol contents, which are related to excessive sunlight energy dissipation as heat, and photoprotection. Accordingly, antioxidant strategies varied according to the gall-inducing species and to the host plant species. The distinctive investments in carotenoid and/or in polyphenol concentrations in the studied galls seemed to be peculiar mechanisms to maintain oxidative homeostasis. These mechanisms were determined both by the stimuli of the gall-inducing organism and by the intrinsic physiological features of the host plant species. Therefore, the roles of both associated organisms in host plant-galling organisms systems over gall metabolism is attested.

Reacquisition of New Meristematic Sites Determines the Development of a New Organ, the Cecidomyiidae Gall on Copaifera langsdorffii Desf. (Fabaceae).

The development of gall shapes has been attributed to the feeding behavior of the galling insects and how the host tissues react to galling stimuli, which ultimately culminate in a variable set of structural responses. A superhost of galling herbivores, Copaifera langsdorffii, hosts a bizarre "horn-shaped" leaflet gall morphotype induced by an unidentified species of Diptera: Cecidomyiidae. By studying the development of this gall morphotype under the anatomical and physiological perspectives, we demonstrate the symptoms of the Cecidomyiidae manipulation over plant tissues, toward the cell redifferentiation and tissue neoformation. The most prominent feature of this gall is the shifting in shape from growth and development phase toward maturation, which imply in metabolites accumulation detected by histochemical tests in meristem-like group of cells within gall structure. We hypothesize that the development of complex galls, such as the horn-shaped demands the reacquisition of cell meristematic competence. Also, as mature galls are green, their photosynthetic activity should be sufficient for their oxygenation, thus compensating the low gas diffusion through the compacted gall parenchyma. We currently conclude that the galling Cecidomyiidae triggers the establishment of new sites of meristematic tissues, which are ultimately responsible for shifting from the young conical to the mature horn-shaped gall morphotype. Accordingly, the conservative photosynthesis activity in gall site maintains tissue homeostasis by avoiding hypoxia and hipercarbia in the highly compacted gall tissues.

Sink Status and Photosynthetic Rate of the Leaflet Galls Induced by Bystracoccus mataybae (Eriococcidae) on Matayba guianensis (Sapindaceae).

The galling insect Bystracoccus mataybae (Eriococcidae) induces green and intralaminar galls on leaflets of Matayba guianensis (Sapindaceae), and promotes a high oxidative stress in host plant tissues. This biotic stress is assumed by the histochemical detection of hydrogen peroxide, a reactive oxygen species (ROS), whose production alters gall physiology. Thus, we hypothesize that high levels of nutrients are accumulated during gall development in response to a local maintenance of photosynthesis and to the galling insect activity. Moreover, the maintenance of low levels of photosynthesis may guarantee O2 production and CO2 consumption, as well as may avoid hypoxia and hypercarbia in gall tissues. To access the photosynthesis performance, the distribution of chlorophyllous tissues and the photochemical and carboxylation rates in gall tissues were analyzed. In addition, histochemical tests for hydrogen peroxide and phenolic derivatives were performed to confirm the biotic stress, and set the possible sites where stress dissipation occurs. The contents of sugars and nitrogen were evaluated to quantify the gall sink. Currently, we assume that the homeostasis in gall tissues is ruptured by the oxidative stress promoted by the galling insect activity. Thus, to supply the demands of gall metabolism, the levels of water-soluble polysaccharides and starch increase in gall tissues. The low values of maximum quantum efficiency of PSII (Fv/Fm) indicate a low photosynthetic performance in gall tissues. In addition, the decrease of PSII operating efficiency, (F'm-F')/F'm, and Rfd (instantaneous fluorescence decline ratio in light, to measure tissue vitality) demonstrate that the tissues of B. mataybae galls are more susceptible to damage caused by stressors than the non-galled tissues. Thus, the high oxidative stress in gall developmental sites is dissipated not only by the accumulation of phenolic derivatives in the protoplast, but also of lignins in the walls of neoformed sclereids.

Developmental anatomy and immunocytochemistry reveal the neo-ontogenesis of the leaf tissues of Psidium myrtoides (Myrtaceae) towards the globoid galls of Nothotrioza myrtoidis (Triozidae).

KEY MESSAGE: The temporal balance between hyperplasia and hypertrophy, and the new functions of different cell lineages led to cell transformations in a centrifugal gradient that determines the gall globoid shape. Plant galls develop by the redifferentiation of new cell types originated from those of the host plants, with new functional and structural designs related to the composition of cell walls and cell contents. Variations in cell wall composition have just started to be explored with the perspective of gall development, and are herein related to the histochemical gradients previously detected on Psidium myrtoides galls. Young and mature leaves of P. myrtoides and galls of Nothotrioza myrtoidis at different developmental stages were analysed using anatomical, cytometrical and immunocytochemical approaches. The gall parenchyma presents transformations in the size and shape of the cells in distinct tissue layers, and variations of pectin and protein domains in cell walls. The temporal balance between tissue hyperplasia and cell hypertrophy, and the new functions of different cell lineages led to cell transformations in a centrifugal gradient, which determines the globoid shape of the gall. The distribution of cell wall epitopes affected cell wall flexibility and rigidity, towards gall maturation. By senescence, it provided functional stability for the outer cortical parenchyma. The detection of the demethylesterified homogalacturonans (HGAs) denoted the activity of the pectin methylesterases (PMEs) during the senescent phase, and was a novel time-based detection linked to the increased rigidity of the cell walls, and to the gall opening. Current investigation firstly reports the influence of immunocytochemistry of plant cell walls over the development of leaf tissues, determining their neo-ontogenesis towards a new phenotype, i.e., the globoid gall morphotype.

Synchronism between Aspidosperma macrocarpon Apocynaceae resources allocation and the establishment of the gall inducer Pseudophacopteron sp. Hemiptera: Psylloidea / Sincronismo de Aspidosperma macrocarpon Apocynaceae entre la asignación de recursos y el establecimiento del inductor de agallas Pseudophacopteron sp. Hemiptera: Psylloidea

The joint interpretation of phenology and nutritional metabolism provides important data on plant tissues reactivity and the period of gall induction. A population of Aspidosperma macrocarpon Apocynaceae with leaf galls induced by a Pseudophacopteron sp. Psylloidea was studied in Goiás state, Brazil. Assuming the morphological similarity between host leaves and intralaminar galls, a gradient from non-galled leaves towards galls should be generated, establishing a morpho-physiological continuum. The phenology, infestation of galls, and the carbohydrate and nitrogen contents were monthly evaluated in 10-20 individuals, from September 2009 to September 2010. Our objective was to analyze the nutritional status and the establishment of a physiological continuum between the galls and the non-galled leaves of A. macrocarpon. The period of leaf flushing coincided with the highest levels of nitrogen allocated to the new leaves, and to the lowest levels of carbohydrates. The nutrients were previously consumed by the growing leaves, by the time of gall induction. The levels of carbohydrates were higher in galls than in non-galled leaves in time-based analyses, which indicateed their potential sink functionality. The leaves were infested in October, galls developed along the year, and gall senescence took place from March to September, together with host leaves. This first senescent leaves caused insect mortality. The higher availability of nutrients at the moment of gall induction was demonstrated and seems to be important not only for the establishment of the galling insect but also for the responsiveness of the host plant tissues.

La interpretación conjunta de la fenología y el metabolismo nutricional proporciona datos importantes sobre la reacción de tejidos vegetales y el período de inducción de agallas. En el estado de Goiás, Brasil se estudió una población de Aspidosperma macrocarpon Apocynaceae con agallas foliares inducidas por un Pseudophacopteron sp. Psylloidea. Suponiendo que la similitud morfológica entre las hojas del hospedero y las agallas intralaminares genera un gradiente de hojas sin agallas hacia hojas con agallas, se establece un continuo morfo-fisiológico. La fenología, la infestación de las agallas y el contenido de carbohidratos y nitrógeno se evaluaron mensualmente en 10-20 individuos, de septiembre 2009 a septiembre 2010. El objetivo fue analizar el estado nutricional y el establecimiento de un continuo fisiológico entre las hojas que producen agallas y las hojas sin agallas de A. macrocarpon. El período de emergencia de hojas coincidió con los niveles más altos de nitrógeno en las hojas nuevas, y los niveles más bajos de carbohidratos.Los nutrientes se consumieron previamente por las hojas en crecimiento, en el momento de inducción de las agallas. Los niveles de carbohidratos fueron más altos en las hojas con agallas que en aquellas sin agallas, en análisis basados en tiempo, lo que indica su ciclo de funcionalidad potencial. Las hojas hospederas se infestaron en octubre, el desarrollo y la senescencia de las agallas se llevó a cabo entre marzo y septiembre, en las hojas hospederas. Estas primeras hojas senescentes causaron mortalidad de insectos. Hay mayor disponibilidad de nutrientes en el momento de la inducción de las agallas y parece ser importante no sólo para el establecimiento del insecto que produce agallas, sino también para la capacidad de respuesta de los tejidos de la planta hospedera.

Synchronism between Aspidosperma macrocarpon (Apocynaceae) resources allocation and the establishment of the gall inducer Pseudophacopteron sp. (Hemiptera: Psylloidea).

The joint interpretation of phenology and nutritional metabolism provides important data on plant tissues reactivity and the period of gall induction. A population of Aspidosperma macrocarpon (Apocynaceae) with leaf galls induced by a Pseudophacopteron sp. (Psylloidea) was studied in Goiás state, Brazil. Assuming the morphological similarity between host leaves and intralaminar galls, a gradient from non-galled leaves towards galls should be generated, establishing a morpho-physiological continuum. The phenology, infestation of galls, and the carbohydrate and nitrogen contents were monthly evaluated in 10-20 individuals, from September 2009 to September 2010. Our objective was to analyze the nutritional status and the establishment of a physiological continuum between the galls and the non-galled leaves of A. macrocarpon. The period of leaf flushing coincided with the highest levels of nitrogen allocated to the new leaves, and to the lowest levels of carbohydrates. The nutrients were previously consumed by the growing leaves, by the time of gall induction. The levels of carbohydrates were higher in galls than in non-galled leaves in time-based analyses, which indicateed their potential sink functionality. The leaves were infested in October, galls developed along the year, and gall senescence took place from March to September, together with host leaves. This first senescent leaves caused insect mortality. The higher availability of nutrients at the moment of gall induction was demonstrated and seems to be important not only for the establishment of the galling insect but also for the responsiveness of the host plant tissues.