Structural patterns of Lepidoptera galls and the case of Andescecidium parrai (Cecidosidae) galls on Schinus polygama (Anacardiaceae).

Gall anatomical and metabolic peculiarities are determined by the feeding habit of the gall inducer, but develop under the constraints of the host plants. The chewing habit of the Lepidoptera larvae imposes a high impact on the host plant cells, and supposedly drives peculiar structural and histochemical patterns. So, our starting point was the search of such patterns in literature, and the test of these traits on the Andescecidium parrai (Cecidosidae)-Schinus polygama (Anacardiaceae) system, as a case study in Chilean flora. The literature on the structure of lepidopteran galls in the temperate and tropical regions comprises 13 works, describing stems as the most frequent host organs, followed by leaves, buds, and flowers. As common structural traits of Lepidoptera galls, the literature converge in describing the processes of cell hypertrophy and hyperplasia, resulting in a variable number of common storage parenchyma layers, interspersed by the redifferentiated sclerenchyma, vascular, and typical nutritive cells around the larval chamber. These nutritive cells accumulate lipids and proteins, which support the lepidopteran larvae nutrition. As expected, the A. parrai galls follow the patterns herein described for the lepidoptera-induced galls, but with peculiarities associated with its host organ. Even though the Lepidoptera galls have destructive mouthparts and can induce large and complex galls, they cannot alter important conservative features of their hosts' organs.

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.

Nicotiana benthamiana as a model for studying Cryptococcus-plant interaction.

Cryptococcus gattii, an environmental yeast isolated from plants, is one of the agents of cryptococcosis. Here, we aimed to develop a plant model to study C. gattii-plant interaction, since it is unclear how it affects the plant and the yeast. We tested three inoculation methods (scarification, infiltration, and abrasion) in three plant species: Arabidopsis thaliana, Nicotiana tabacum, and N. benthamiana. Cryptococcus gattii was able to grow in all three models, with a peak of yeast cell burden after 7 days, without any pathological effects. Furthermore, the fungal burden was reduced later, confirming that C. gattii is not a phytopathogen. Cryptococcus gattii proliferation was higher in N. benthamiana, which presented an increase in the hydrogen peroxide content, antioxidant system activity, and indoleacetic acid (IAA) production. Cryptococcus gattii colonies recovered from N. benthamiana presented lower ergosterol content, reduced capsule, and increased growth rate in vitro and inside macrophages. In vitro, IAA altered C. gattii morphology and susceptibility to antifungal drugs. We hypothesize that C. gattii can temporarily colonize plant living tissues, which can be a potential reservoir of yeast virulence, with further dissemination to the environment, birds, and mammals. In conclusion, N. benthamiana is suitable for studying C. gattii-plant interaction.

Complex meristematic activity induced by Eucecidoses minutanus on Schinus engleri turns shoots into galls.

PREMISE: Gall-inducing organisms change the development of their host plant organs, resulting in ontogenetic patterns not observed in the non-galled plants. Distinct taxa induce galls on Schinus spp., manipulating meristematic patterns in the host plant in distinct ways. Here we report ontogenetic novelties induced in the lateral buds of S. engleri by Eucecidoses minutanus, a Cecidosidae, whose galls have been poorly understood. METHODS: The anatomy, histochemistry, and histometry of galls in distinct phases of development, non-galled buds, and stems of Schinus engleri were analyzed in parallel with the instars of E. minutanus to detail the morphogenetic changes in the host with each larval stage. RESULTS: Ontogenetic phases of the galls were intricately associated with larval development. First and second-instar larvae induced pericycle and pith cells to dedifferentiate into the gall inner meristem, where hyperplasia and cell hypertrophy characterized the growth and development phase of the gall. The innermost layers were lipid-rich nutritive cells that lined the larval chamber. Additional vascular bundle rows were produced in young galls. Third and fourth instar-larvae were associated with the gall maturation phase: centripetal lignification of the outer parenchyma cell layers, epidermal stratification, and activation of a cambium-like meristem (CLM). The CLM activity resulted in new layers of nutritive cells that differentiated inward as the first layers of nutritive cells were consumed by E. minutanus larvae, and, also, in more parenchyma cell layers that formed outward. All tissues between the innermost layer of nutritive tissue that surround the gall chamber and the outermost layer of the dermal system that externally covers the gall form the gall wall, and increased in thickness until the end of gall maturation. CONCLUSIONS: E. minutanus induces a structurally complex globoid stem gall, modifying all host plant tissues and stimulating a novel meristematic pattern in S. engleri. The gall developmental stages are each related to specific gall-inducing instars, as gall development progresses according to the development of E. minutanus.

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.

Photosynthetic heat tolerance in plants with different foliar water -uptake strategies.

PREMISE: The distribution and even the survival of plant species are influenced by temperature. In an old climatically buffered infertile landscape (OCBIL) in Brazil, we previously characterized different strategies for foliar water uptake (FWU). It is possible that photosystem II tolerance to heat and excessive light intensity varies among species with different FWU capacities. METHODS: The relationship between FWU, photoinhibition, and thermotolerance was investigated in seven species from this ecosystem. RESULTS: The species with slow water absorption and high water absorption are those that presented less photoinhibition. Contrastingly, the species that have fast and low water absorption presented greater thermotolerance when their leaves are totally hydrated. However, when there is greater leaf dehydration, the most thermotolerant species were those with slow but high water absorption. CONCLUSIONS: Foliar water uptake is an important trait for plants to tolerate excessive light intensity and higher temperatures. Plants in this OCBIL may be differentially affected by future global warming, and the best strategy to deal with this expected climate change is with slow and high absorption of water.

The Neglected Reverse Water Pathway: Atmosphere-Plant-Soil Continuum.

The soil-plant-atmosphere continuum (SPAC) describes the continuous water movement from soil via plants to atmosphere. Here, we propose to name the reverse water pathway, driven by foliar water uptake, the atmosphere-plant-soil continuum (APSC). We highlight the different hydraulic resistances this reverse water movement has to overcome.

Cytological attributes of storage tissues in nematode and eriophyid galls: pectin and hemicellulose functional insights.

Cell walls and protoplast may work together or distinctly in the establishment of the functional profiles of gall tissue compartments. This presumption is herein evaluated in three gall systems by immunocytochemical and ultrastructural analyses. The common storage tissues (CSTs) of leaf galls induced by Eriophyidae on Miconia ibaguensis leaves and by Ditylenchus gallaeformans on M. ibaguensis and M. albicans have rigid and porous cell walls due to their composition of pectins. Hemicelluloses in CST cell walls are scarcer when compared to the cell walls of the control leaves, being functionally compensated by rigid pectate gels. The typical nutritive tissues (TNTs) in galls induced by Ditylenchus gallaeformans are similar to promeristematic and secretory cells regarding their enriched cytoplasm, several mitochondria, and proplastids, as well as multivesicular and prolamellar bodies in cell membranes. The cytological features of the feeding cells of Eriophyidae galls indicate that they are not as metabolically active as the cells of the TNT in nematode galls. However, their cell wall composition suggests more plasticity and porosity than the cells of the TNT, which can compensate the less production of nutrients with more transport. The ultrastructural and immunocytochemical profiles of CST cells reveal functional similarities, which are independent of the taxa of the gall inducer or of the host plant. Despite their analogous functionalities, the protoplast and cell wall features of TNT cells of nematode galls and of the feeding cells of the Eriophyidae galls are distinct, and work out through different strategies toward keeping gall developmental site active.

Vallissiana universitaria (Lepidoptera: Gracillariidae): a new genus and species of leaf-mining moth associated with Erythroxylum (Erythroxylaceae) in the Atlantic Forest of Brazil.

Vallissiana universitaria Pereira Arévalo, a new genus and species of leaf-miner moth (Gracillariidae: Gracillariinae) is described and illustrated with the aid of optical and scanning electron microscopy, including adults, larva, pupa and the mine. Its monophyletic status is confirmed within the subfamily based on a DNA barcode CoI tree. The immature stages are associated with Erythroxylum argentinum O. E. Schulz (Erythroxylaceae) and four larval instars are found, all forming a round blotch mine from the beginning of ontogeny. The first two instars are sap-feeders, using only the epidermal cells, whereas the last two are tissue-feeders, mining the parenchyma cells. Pupation occurs inside the leaf mine within a flimsy, silk-made cocoon. This is the third endemic genus of gracillariid moths described from the Atlantic Forest of Brazil and the first associated with Erythroxylum P. Browne. Characteristics found on the forewing and in the last abdominal segments of the adult were determinant for the proposition of the new genus. The CoI tree indicated that it is closely related to Aspilapteryx, while this genus was recovered as polyphyletic in the analyses. Morphological evidence supports this polyphyly. Consequently, Sabulopteryx Triberti, 1985, stat. nov. is considered a valid genus.

Foliar water-uptake strategies are related to leaf water status and gas exchange in plants from a ferruginous rupestrian field.

PREMISE: Fog is a frequent event in Brazilian rupestrian field and plays an important role in the physiology of several plant species. Foliar water uptake (FWU) of fog may be fast or slow depending on the species. However, fog water may negatively affect CO2 assimilation. Thus, the interference in the water and carbon balance as a result of different strategies of FWU was evaluated to verify whether fog may mitigate possible water deficit in leaves. METHODS: Four plant species with different FWU strategies were studied in a ferruginous rupestrian field with frequent fog. Gas exchange and water potential were measured before dawn and at midday during the dry and rainy seasons, separating foggy from non-foggy days during the dry season. RESULTS: The FWU speed negatively influences CO2 assimilation in the dry season, possibly because of its negative relationship with stomatal conductance, since reduced stomatal aperture impairs carbon entrance. Fog presence increased leaf water potential both in early morning and midday during the dry season. However, during the rainy season, the values of leaf water potential were lower at midday, than during the dry season with fog at midday, which favors leaf gas exchanges. CONCLUSIONS: FWU interferes negatively, but briefly with CO2 assimilation. Nevertheless, FWU prevents water loss through transpiration and increases the water status of plants in the dry season. That is, FWU results in a compensation between CO2 assimilation and foliar hydration, which, in fact, is beneficial to the plants of this ecosystem.

Spatiotemporal variation in phenolic levels in galls of calophyids on Schinus polygama (Anacardiaceae).

The expression of plant secondary metabolism is strongly controlled by plant both in time and space. Although the variation of secondary metabolites, such as soluble and structural phenolics (e.g., lignins), has been largely observed in gall-inducing insects, and compared to their non-galled host organs, only a few datasets recording such variation are available. Accordingly, the relative importance of spatiotemporal variability in phenolic contents, and the influence of gall developmental stages on the original composition of host organs are poorly discussed. To address this knowledge gap, we histochemically determined the sites of polyphenol and lignin accumulation, and the polyphenol contents in three developmental stages of two calophyid galls and their correspondent host organs. Current results indicate that the compartmentalization of phenolics and lignins on Schinus polygama (Cav.) Cabrera follows a similar pattern in the two-calophyid galls, accumulating in the outer (the external tissue layers) and in the inner tissue compartments (the cell layers in contact with the gall chamber). The non-accumulation in the median compartment (median parenchyma layers of gall wall with vascular bundles, where gall inducer feeds) is important for the inducer, because its mouth apparatus enter in contact with the cells of this compartment. Also, the concentration of phenolics has opposite dynamics, decreasing in leaf galls and increasing in stem galls, in temporal scale, i.e., from maturation toward senescence. The concentration of phenolics in non-galled host organs, and in both galls indicated the extended phenotype of Calophya rubra (Blanchard) and C. mammifex Burckhardt & Basset (Hemiptera: Sternorrhyncha: Psylloidea: Calophyidae) over the same host plant metabolic potentiality.

A redescription of Antispastis clarkei Pastrana (Lepidoptera, Glyphipterigidae) immature stages, with notes on the life history and phylogenetic placement of the genus

Abstract Antispastis Meyrick, 1926 is a poorly known genus of leaf-mining micromoths endemic to the Neotropics, with still uncertain taxonomic position within the Yponomeutoidea. In the present study, the egg, larva and pupa of A. clarkei Pastrana, previously known only from Argentina, are described and illustrated with the aid of optical and scanning electron microscopy. Data on life history, including histology of the mine, are also provided. Family placement of the genus is reassessed based on comparison of morphology and DNA sequences with closely related lineages. The larvae form blotch mines on the upper surface of Solanum L. (Solanaceae) leaves, feeding on palisade parenchyma in all instars. Pupation occurs outside the mine, in an inverted basket-like, large-meshed cocoon constructed on the plant surface. DNA analysis of Cytochrome oxidase I gene of A. clarkei revealed interspecific differences averaging 10% with A. xylophragma, which provided species separation matching morphological differences. Antispastis was closely related phylogenetically to Digitivalva, clustering in the Acrolepiinae together with the genera Acrolepia and Acrolepiopsis, and ultimately placed within Glyphipterigidae. The geographical distribution of A. clarkei is expanded to the Southern Atlantic forest (Rio Grande do Sul and Paraná states, Brazil).

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.

Preventing False Negatives for Histochemical Detection of Phenolics and Lignins in PEG-Embedded Plant Tissues.

Polyethylene glycol (PEG) is a low-cost and advantageous embedding medium, which maintains the majority of cell contents unaltered during the embedding process. Some hard or complex plant materials are better embedded in PEG than in other usual embedding media. However, the histochemical tests for phenolics and lignins in PEG-embedded plant tissues commonly result in false negatives. We hypothesize that these false negatives should be prevented by the use of distinct fixatives, which should avoid the bonds between PEG and phenols. Novel protocols for phenolics and flavanols detection are efficiently tested, with fixation of the samples in ferrous sulfate and formalin or in caffeine and sodium benzoate, respectively. The differentiation of lignin types is possible in safranin-stained sections observed under fluorescence. The Maule's test faultlessly distinguishes syringyl-rich from guaiacyl- and hydroxyphenyl-rich lignins in PEG-embedded material under light microscopy. Current hypothesis is corroborated, that is, the adequate fixation solves the false-negative results, and the new proposed protocols fill up some gaps on the detection of phenolics and lignins.

Multivesicular bodies differentiate exclusively in nutritive fast-dividing cells in Marcetia taxifolia galls.

Marcetia taxifolia (A. St.-Hil.) DC. hosts two gall morphotypes, a pistil-shaped gall induced by a Cecidomyiidae (Diptera) and a fusiform stem gall induced by a Lepidoptera. The cytological study of these galls aimed to answer how the difference in nutritive tissues of Diptera and Lepidoptera galls could be explained on cytological basis. The nutritive tissues of lepidopteran galls have a fast-dividing cell zone, the storage nutritive tissue, which replaces the cells of the typical nutritive tissue, where the larvae feed. The differentiation of multivesicular bodies in the plasma membrane occurred exclusively in these fast-dividing cells of the lepidopteran galls, evidencing the meristematic condition of such tissue. The accumulation of reactive oxygen species (ROS) analyzed in situ in the nutritive cells is not sufficient to induce programmed cell death (PCD), as the cells of M. taxifolia have plastoglobules and accumulate polyphenols and terpenoids, which are diagnostic defenses against oxidative stress. The two taxa of galling insects have different nutritional requirements, thus inducing specific cytoplasm-enriched cells on their nutritive tissues.

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.

Efficiency of the Polyethylene-Glycol (PEG) Embedding Medium for Plant Histochemistry.

Histochemical analyses in plants are commonly performed on hand-made sections of fresh materials. The disadvantages of embedding in historesin, paraffin or paraplast® are the alterations to cellular contents, the high costs and few evident results, depending on the test. Polyethylene-glycol (PEG), as a low cost, hydrophilic medium that maintains most of the cellular features similar to fresh conditions, may be useful for obtaining good histochemical results in thinner and homogeneous sections. The current study aimed to compare the efficiency of PEG as an embedding medium for histochemical analyses of primary and secondary metabolites accumulation. Using hand-made sections of fresh samples (T1) as a comparison, we tested the influence of the use of Karnovsky's solution as a fixative (T2) versus embedding in PEG (T3). The samples herein analyzed comprise leaves, stems, seeds and insect galls of different plant species. Neither the Karnovsky's fixative nor the embedding in PEG altered the histochemical results for starch, lipids, terpenoids, proteins and reducing sugars in T1, T2, and T3. However, PEG binds to phenols, such as tannins, flavonoids and lignins, thereby presenting false negatives in T3.

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.