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Structural and physiological leaf traits and their plasticity were compared in the hemiepiphyte Vanilla phaeantha . This species grows along a phorophyte reaching different understorey positions and exhibiting diverse responses to environment changes. We analysed three height strata above the ground, establishing a light gradient, and considering seasonal water fluctuations. The upper leaves had higher area and mass and were less pigmented. The dry season induced a reduction of approximately 2h of stomatal opening over the diel 24h crassulacean acid metabolism (CAM) cycle in the leaves at all understorey positions. The leaves more exposed to sunlight were larger with higher titratable acidity during the rainy season, while the leaves near the ground maintained the same rates of stomatal conductance and nocturnal acidification between seasons, with lowest values of carbon isotopes in the rainy season. Our research showed that some structural leaf traits (such as specific leaf mass, biomass, and saturated water content) are sensitive to variation in understorey position. In contrast, other physiological traits (stomatal conductance, transpiration, and fluorescence parameters) are more sensitive to seasonal variations. The results are a novelty in assessing the variation of CAM along the same plant in a height gradient and under field conditions.

In response to the restrictions imposed by their epiphytic habit, orchids have developed structural traits that allow greater efficiency in water uptake and use, such as a complex adventitious root system with velamen. The composition of cell wall of this specialized epidermis can be altered according to the substrate to which it is fixed, influencing wall permeability, absorption, and storage of water in roots. The current study aimed to evaluate the cell wall composition of adventitious roots of Vanilla phaeantha (Orchidaceae) that grow attached to the phorophyte, fixed in the soil, or hung free. Immunocytochemical analyses were used to determine the protein, hemicellulose, and pectin composition of the cell walls of aerial and terrestrial roots. We observed that pectins are present in the different tissues of the aerial roots, while in the terrestrial roots, they are concentrated in the cortical parenchyma. The deposition of xyloglucans, extensins, and arabinogalactans was greater in the epidermis of the free side of the roots attached to the phorophyte. The strong labeling of pectins in aerial roots may be related to the influx of water and nutrients, which are generally scarce in this environment. The arrangement of hemicelluloses and proteins with the pectins may be associated with increased cell rigidity and sustainability, a feature of interest for the aerial roots. In summary, the habit of roots can interfere with the non-cellulosic composition of the cell walls of V. phaeantha, possibly related to changes in cell functionality., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Epiphytic and rupicolous plants inhabit environments with limited water resources. Such plants commonly use Crassulacean Acid Metabolism (CAM), a photosynthetic pathway that accumulates organic acids in cell vacuoles at night, so reducing their leaf water potential and favouring water absorption. Foliar water uptake (FWU) aids plant survival during drought events in environments with high water deficits. We hypothesized that FWU represents a strategy employed by epiphytic and rupicolous orchids for water acquisition and that CAM will favour increased water absorption. We examined 6 epiphyte, 4 terrestrial and 6 rupicolous orchids that use C3 (n = 9) or CAM (n = 7) pathways. Five individuals per species were used to evaluate FWU, structural characteristics and leaf water balance. Rupicolous species with C3 metabolism had higher FWU than other species. FWU (C max and k) could be related to succulence, SLM and leaf RWC. The results indicated that high orchid leaf densities favoured FWU, as area available for water storage increases with leaf density. Structural characteristics linked to water storage (e.g. high RWC, succulence), on the other hand, could limit leaf water absorption by favouring high internal leaf water potentials. Epiphytic, rupicolous and terrestrial orchids showed FWU. Rupicolous species had high levels of FWU, probably through absorption from mist. However, succulence in plants with CAM appears to mitigate FWU., (© 2022 German Society for Plant Sciences, Royal Botanical Society of the Netherlands.)

The aerial environment appears to structurally modify roots, which frequently show specializations for absorbing water and nutrients. Among those specializations are the velamen, a multiseriate epidermis generally composed of dead mature cells, and greater degrees of lignification in the endodermis, exodermis, and pith. Vanilla phaeantha is a hemiepiphyte used here as a model of study to determine which root characteristics demonstrate the most plasticity in response to aerial and terrestrial environments. It produces roots growing under three conditions: (1) aerial and free, growing from the highest branches towards the ground; (2) aerial roots attached to the phorophyte; and (3) terrestrial. Samples taken 3 cm from the apices were used to prepare histological slides. The tissues and other anatomical structures were measured and histochemically characterized. The most plastic characteristics were the external periclinal thicknesses of the exodermis and the total area occupied by the aerenchyma lacunae. The free roots were the longest, did not evidence root hairs, and had the largest number of the aerenchyma lacunae; they also evidenced greater thicknesses of the exodermis in contact with the epidermis walls that helped maintain their shapes. Terrestrial roots had root hairs around the entire circumference and intense infestations of mycorrhiza, indicating their involvement in nutrient acquisition. The adhering roots evidenced free regions similar to those of aerial roots, as well as adhering regions showed characteristics similar to terrestrial roots (with root hairs and mycorrhiza infestations)., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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.

The chemical composition of the cell walls strongly affects water permeability and storage in root tissues. Since epiphytic orchids live in a habitat with a highly fluctuating water supply, the root cell walls are functionally important. In the present study, we used histochemistry and immunocytochemistry techniques in order to determine the composition of the cell walls of root tissues of 18 epiphytic species belonging to seven subtribes across the Orchidaceae. The impregnation of lignin in the velamen cells reinforces its function as mechanical support and can facilitate apoplastic flow. Pectins, as well cellulose and lignins, are also essential for the stability and mechanical support of velamen cells. The exodermis and endodermis possess a suberinized lamella and often lignified walls that function as selective barriers to apoplastic flow. Various cortical parenchyma secondary wall thickenings, including phi, reticulated, and uniform, prevent the cortex from collapsing during periods of desiccation. The presence of highly methyl-esterified pectins in the cortical parenchyma facilitates the formation of gels, causing wall loosening and increased porosity, which contributes to water storage and solute transport between cells. Finally, cells with lipid or lignin impregnation in the cortical parenchyma could increase the water flow towards the stele.

The galls induced by Ditylenchus gallaeformans (Nematoda) on leaves of Miconia albicans have unique features when compared to other galls. The nematode colonies are surrounded by nutritive tissues with promeristematic cells, capable of originating new emergences facing the larval chamber, and providing indeterminate growth to these galls. Considering enzyme activity as essential for the translocation of energetic molecules from the common storage tissue (CST) to the typical nutritive tissue (TNT), and the major occurrence of carbohydrates in nematode galls, it was expected that hormones would mediate sink strength relationships by activating enzymes in indeterminate growth regions of the galls. Histochemical, immunocytochemical and quantitative analyses were made in order to demonstrate sites of enzyme activity and hormones, and comparative levels of total soluble sugars, water soluble polysaccharides and starch. The source-sink status, via carbohydrate metabolism, is controlled by the major accumulation of cytokinins in totipotent nutritive cells and new emergences. Thus, reducing sugars, such as glucose and fructose, accumulate in the TNT, where they supply the energy for successive cycles of cell division and for nematode feeding. The histochemical detection of phosphorylase and invertase activities indicates the occurrence of starch catabolism and sucrose transformation into reducing sugars, respectively, in the establishment of a gradient from the CST towards the TNT. Reducing sugars in the TNT are important for the production of new cell walls during the indeterminate growth of the galls, which have increased levels of water-soluble polysaccharides that corroborate such a hypothesis. Functional relationship between plant hormone accumulation, carbohydrate metabolism and cell differentiation in D. gallaeformans-induced galls is attested, providing new insights on cell development and plant metabolism., (© 2019 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

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., Competing Interests: The authors have declared that no competing interests exist.

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.

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 O 2 production and CO 2 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 ( F v / F m ) 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.