Anatomical, histochemical, and developmental approaches reveal the long-term functioning of the floral nectary in Tocoyena formosa (Rubiaceae).

Tocoyena formosa has a persistent floral nectary that continues producing nectar throughout flower and fruit development. This plant also presents an intriguing non-anthetic nectary derived from early-developing floral buds with premature abscised corolla. In this study, we characterize the structure, morphological changes, and functioning of T. formosa floral nectary at different developmental stages. We subdivided the nectary into four categories based on the floral and fruit development stage at which nectar production started: (i) non-anthetic nectary; (ii) anthetic nectary, which follows the regular floral development; (iii) pericarpial nectary, derived from pollinated flowers following fruit development; and (iv) post-anthetic nectary that results from non-pollinated flowers after anthesis. The nectary has a uniseriate epidermis with stomata, nectariferous parenchyma, and vascular bundles, with a predominating phloem at the periphery. The non-anthetic nectary presents immature tissues that release the exudate. The nectary progressively becomes more rigid as the flower and fruit develop. The main nectary changes during flower and fruit development comprised the thickening of the cuticle and epidermal cell walls, formation of cuticular epithelium, and an increase in the abundance of calcium oxalate crystals and phenolic cells near the vascular bundles. Projections of the outer periclinal walls toward the cuticle in the post-anthetic nectary suggest nectar reabsorption. The anatomical changes of the nectary allow it to function for an extended period throughout floral and fruit development. Hence, T. formosa nectary is a bivalent secretory structure that plays a crucial role in the reproductive and defensive interactions of this plant species.

Oil and mucilage idioblasts co-occur in the vegetative organs of Ocotea pulchella (Lauraceae): comparative development, ultrastructure and secretions.

This study compares oil and mucilage idioblasts occurring together in the vegetative organs of Ocotea pulchella, a Lauraceae species. Our focus is specifically on the ontogeny and developmental cytology of these secretory cells. Both types of idioblasts originate from solitary cells located in the fundamental meristem, underlying the protodermis. The growth of both types of idioblasts is asynchronous, with the oil idioblasts developing first, but their initiation is restricted to the early stages of organ development. Mucilaginous idioblasts occur exclusively in the palisade parenchyma, while oil idioblasts are scattered throughout the mesophyll, midrib, and petiole of the leaves. The lamellar secretion of mucilage idioblasts is mostly made up of polysaccharides, while the secretion of oil idioblasts is made up of terpenes and lipids. Cupule occurred only in the oil idioblasts, while suberized layers occurred in both types of cells. We found that immature oil idioblasts that are close to each other fuse; mature mucilage idioblasts have labyrinthine walls arranged in a reticulate pattern; the cells close to the oil idioblasts have a pectin protective layer; and the oil idioblasts have a sheath of phenolic cells. In contrast to previous reports, the two types of secretory idioblasts were recognized during the early stages of their development. The results emphasize the importance of combining optical and electron microscopy methods to observe the ontogenetic, histochemical and ultrastructural changes that occur during the development of the secretory idioblasts. This can help us understand how secreting cells store their secretions and how their walls become specialized.

The putative cannabinoid-secreting trichome of Trema micrantha (L.) Blume (Cannabaceae).

Trema, a genus of the popularly known Cannabaceae, has recently been the subject of cannabinoid bioprospection. T. micrantha is a tree with pharmacological potential widely used in folk medicine. It has two types of glandular trichomes, bulbous and filiform, spread throughout the plant body. Considering the proximity of this species to Cannabis sativa and Trema orientalis, species containing cannabinoids, the glandular trichomes of T. micrantha are also expected to be related to the secretion of these compounds. Thus, this study aims to detail the morphology of secretory trichomes during the synthesis, storing and release of metabolites in T. micrantha. We tested the proposition that they could be a putative type of cannabinoid-secreting gland. Pistillate and staminate flowers and leaves were collected and processed for ontogenic, histochemical, and ultrastructural analyses. Both types of glandular trichomes originate from a protodermal cell. They are putative cannabinoid-secreting sites because: (1) terpene-phenols and, more specifically, cannabinoids were detected in situ; (2) their secretory subcellular apparatus is consistent with that found in C. sativa: modified plastids, polyribosomes, an extensive rough endoplasmic reticulum, and a moniliform smooth endoplasmic reticulum. Plastids and smooth endoplasmic reticulum are involved in the synthesis of terpenes, while the rough endoplasmic reticulum acts in the phenolic synthesis. These substances cross the plasma membrane by exocytosis and are released outside the trichome through cuticle pores. The study of the cell biology of the putative cannabinoid glands can promote the advancement of prospecting for natural products in plants.

Osmophores and petal surface traits in Bignonieae species.

This study characterizes the osmophores and corolla traits in 18 species of Bignonieae Dumort., a Bignoniaceae tribe occurring in the Cerrado, a neotropical savanna in Brazil. To detect osmophore distribution, whole, newly opened flowers were immersed in Neutral Red Solution. Samples from the corolla tube and lobes were also fixed and analyzed micromorphologically, anatomically, and histochemically. The osmophores showed six markedly different distribution patterns that were not clearly associated with histological features. In most species, osmophores comprised papillose secretory epidermises and a few layers of subepidermal parenchyma. Starch grains, lipid droplets, and terpenes were detected in osmophores. An ornamented cuticle, cuticular folds, glandular and non-glandular trichomes, raised stomata and epicuticular wax granules are common traits in the species studied and may be useful in determining the taxonomy of the group. We found that 94% of the species visited by bees had papillose epidermises while the single hummingbird-pollinated species presented a flattened epidermis. Variations in osmophore pattern among species visited by bees, including variations within the same plant genus, are novel finding. Additionally, the Bignonieae species visited by bees presented a textured corolla surface, which has been reported as facilitating bee attachment and movement towards the floral resource. Future studies with a greater number of Bignonieae species and more detailed pollinator behavioral assays may help in the interpretation of the variations in corolla traits and functional relationships between flowers and pollinators.

To each their own! Nectar plasticity within a flower mediates distinct ecological interactions.

Nuptial and extranuptial nectaries are involved in interactions with different animal functional groups. Nectar traits involved in pollination mutualisms are well known. However, we know little about those traits involved in other mutualisms, such as ant-plant interactions, especially when both types of nectaries are in the same plant organ, the flower. Here we investigated if when two types of nectaries are exploited by distinct functional groups of floral visitors, even being within the same plant organ, the nectar secreted presents distinct features that fit animal requirements. We compared nectar secretion dynamics, floral visitors and nectar chemical composition of both nuptial and extranuptial nectaries in natural populations of the liana Amphilophium mansoanum (Bignoniaceae). For that we characterized nectar sugar, amino acid and specialized metabolite composition by high-performance liquid chromatography. Nuptial nectaries were visited by three medium- and large-sized bee species and extranuptial nectaries were visited mainly by ants, but also by cockroaches, wasps and flies. Nuptial and extranuptial nectar differed regarding volume, concentration, milligrams of sugars per flower and secretion dynamics. Nuptial nectar was sucrose-dominated, with high amounts of γ-aminobutyric acid and ß-aminobutyric acid and with theophylline-like alkaloid, which were all exclusive of nuptial nectar. Whereas extranuptial nectar was hexose-rich, had a richer and less variable amino acid chemical profile, with high amounts of serine and alanine amino acids and with higher amounts of the specialized metabolite tyramine. The nectar traits from nuptial and extranuptial nectaries differ in energy amount and nutritional value, as well as in neuroactive specialized metabolites. These differences seem to match floral visitors' requirements, since they exclusively consume one of the two nectar types and may be exerting selective pressures on the composition of the respective resources of interest.

Intraspecific variation in ultrastructure and secretion of the resin canals in Anacardium humile (Anacardiaceae).

The study combines a range of light and electron microscopy methods to access variation in secretion and ultrastructure in the secretory canals in the above- and belowground stems of Anacardium humile, which here serves as a model system. The aboveground stem canals show epithelial cells with ultrastructural characteristics typical of cells active in secretion, while in the belowground stems, the subcellular characteristics are typical of cells with low rates of metabolism. The secretory canals of the belowground stems show uniformity in size and shape, a large central vacuole, a cytoplasm reduced to a thin layer at the cell periphery, and a reduced population of organelles. The aboveground stem canals had voluminous nuclei with evident nucleoli, a very dense cytoplasm with free ribosomes, polyribosomes, mitochondria with developed cristae, and ellipsoid plastids with electron-opaque droplets surrounded by a periplastid reticulum. The vacuoles were of different sizes and often had membranous contents and the dictyosomes were very developed with dilated ends to the cisternae, rough endoplasmic reticulum, and numerous vesicles. The results show that particularities in above- and belowground environment have significant implications for ultrastructural morphology and functioning of secretory canals in the stems of A. humile.

Apoplasmic barrier in the extrafloral nectary of Citharexylum myrianthum (Verbenaceae).

MAIN CONCLUSION: The cytological changes underlying the formation of an apoplasmic barrier in the multi-layered extrafloral nectaries of Citharexylum myrianthum are compatible with the synthesis, transport and deposition of suberin. In terms of ontogenesis and function, the intermediate layers of these nectaries are homologous with the stalks of nectar-secreting trichomes. Anticlinal cell wall impregnations are common in trichomatic nectaries and their functions as endodermis-like barriers have been discussed because of possible direct effects on the nectary physiology, mainly in the nectar secretion and resorption. However, the cytological events linked to nectary wall impregnations remain little explored. This study documents the ontogenesis and the fine structure of the EFN cells, and cytological events linked to the wall impregnations of multi-layered extrafloral nectaries (EFNs) in Citharexylum myrianthum Cham. (Verbenaceae). EFNs are patelliform, and differentiated into (a) a multicellular foot, which is compound in structure and vascularised with phloem strands, (b) a bi-layered intermediate region with thickened cell walls and (c) a single-layered secretory region with palisade-like cells. EFNs are protodermal in origin, starting with a single protodermal cell and ending with the complex, multi-layered structure. The cell wall impregnations first appear in the very young EFN and increase towards maturity. Lipid patches (assumed to be suberin) are deposited on the inner faces of the primary walls, first along the anticlinal walls and then extend to the periclinal walls. On both walls, plasmodesmata remain apparently intact during the maturation of the EFNs. In the peripheral cytoplasm there are abundant polymorphic plastids, well-developed Golgi bodies often close to rough endoplasmic reticulum profiles, mitochondria and polyribosomes. Cytological events linked to the wall impregnations are consistent with suberin synthesis, transport and deposition. Our findings offer new insights into the structure-properties of specialised nectary cell walls and so should contribute to our knowledge of the physiological and protective roles of this structure in nectar glands.

Epicormic bud protection traits vary along a latitudinal gradient in a neotropical savanna.

Regrowth via production of epicormic shoots is an important strategy for many woody plants after environmental disturbances such as fire, drought, and herbivory. Populations spreading across a broad latitudinal gradient offer opportunities to investigate if essential traits vary with heterogenous environmental conditions, such as in savanna ecosystems. This information can help us predict plant responses to climate change. Here, we evaluated if epicormic bud protection traits varied among populations of three focal savanna species (Miconia albicans, Solanum lycocarpum, and Zeyheria montana) that have a wide distribution and grow under variable climatic conditions. We randomly sampled 225 individuals over five spatially independent sites (7°, 10°, 15°, 18°, and 24° S) in Brazil, totaling 15 individuals per species per area. We analyzed anatomical transverse sections of five buds per species per area to assess the relative area occupied by crystal and phenolic idioblasts, the thickness of the trichome boundary layer, and to test if these traits were associated with climatic conditions. The buds were protected by cataphylls and composed of a variable number of undeveloped leaves enveloping the shoot apex. For M. albicans, we found an association between maximum temperature and both phenolic idioblasts and trichome boundary layer, but no association with crystal idioblasts. In S. lycocarpum, only the trichome boundary layer was associated with maximum temperature plus high radiation. Z. montana showed no variation. Combination of two or more traits can lead to the development of adaptative strategies to different climatic conditions. We present for the first time an analysis of epicormic bud traits in plant populations occurring in an extensive latitudinal gradient and shed light on how maximum temperature is associated with these traits, contributing to a better understanding of plant resprouting capabilities in widespread savanna plant species.

Anatomical and ultrastructural studies reveal temporal and spatial variation in the oil production in leaves of the diesel tree (Copaifera langsdorffii, Leguminosae).

The oily resin produced by Copaifera langsdorffii, commonly called oil of copaiba, is widely exploited by the drug, cosmetic, and biodiesel industries. The distribution of oily secretory cavities and canals (secretory spaces) over the vegetative body characterizes this species. Oil is stored inside the lumen of the secretory spaces and only reaches the organ surface after injuries. Nonetheless, translucent oily deposits occur on the adaxial surface of intact young leaves. In this study, we searched for further sources of oil production in C. langsdorffii leaves in addition to the well-known secretory cavities and investigated the mechanisms of secretion. Leaves in different developmental stages were collected from adult plants and processed for studies on light and transmission electron microscopies. The primary finding of this study was the involvement of the chlorenchyma cells in lipid biosynthesis, in addition to the secretory cavities. The secretory activity of cavities and chlorenchyma cells overlapped in young leaves. Ultrastructurally, secretory cavity cells exhibited abundant smooth endoplasmic reticulum profiles and oleoplasts, whereas the chlorenchyma cells had large chloroplasts with oil inclusions. Our data suggest that the oily material on the leaf surface arose from the chlorenchyma and was transported via the apoplast. These findings open new avenues for understanding oil biosynthesis regulation in mesophyll cells and planning of future strategies for the biotechnological application of C. langsdorffii leaves.

Lip morphology and ultrastructure of osmophores in Cyclopogon (Orchidaceae) reveal a degree of morphological differentiation among species.

Floral fragrances play an important role in pollinator attraction; they serve as signals for primary rewards such as nectar. The presence of osmophores (a specialized glandular tissue that produces fragrance) in the terrestrial orchid Cyclopogon has only been described in a single species. There are fragrant and odorless species within this genus, but no data on the similarities or differences between them has been published. Here, we present new data on the lip morphology and ultrastructure from eight Cyclopogon species to better understand the mechanisms of pollinator attraction of this group. To achieve this goal, we used light, scanning, and transmission electron microscopy. Five out of eight species presented globular trichomes associated with fragrance production on the abaxial side of the labellum. Together with subepidermal cells, they compose the secretory tissue. Despite the absence of globular trichomes, C. apricus also presented cells associated with fragrance production. Lipid droplets, plastids containing plastoglobuli, and starch were indicators of secretory activity present in these species. This same pattern of fragrance production and release, associated with flower morphology, may result in the restriction of pollination by halictid bees.

Megagametophyte development and female sterility in Maytenus obtusifolia Mart. (Celastraceae).

Reproduction in flowering plants is closely related to the megagametophyte, since the megagametophyte is involved in pollen tube reception and contains the two female gametes-egg cell and central cell. Previous conventional light microscopy methods have shown that female sterility in perfect flowers of Maytenus obtusifolia is associated with the occurrence of sterile ovules whose megagametophytes have hypertrophied synergids. Here, using transmission electron microscopy and cytochemical methods, we compare the megagametophytes in fertile and sterile ovules from perfect and pistillate flowers, and investigate the cellular events that result in the degradation of the megagametophyte cells from sterile ovules. In fertile ovules of perfect and pistillate flowers, mature megagametophytes have two synergids, egg cell and central cell. In fertile ovules, the synergids present an extensive rough endoplasmic reticulum (RER) profile, large populations of mitochondria, when compared to egg cells, vesicles, Golgi bodies, plastids and a nucleus with heterochromatin. Besides that, the egg cell has a small population of organelles and the central cell exhibits cytoplasm with free ribosomes, RER, vesicles originating from the RER, Golgi bodies and oil inclusions. In mature megagametophytes from sterile ovules of perfect and pistillate flowers, massive autophagy occurs by tonoplast rupture promoting hydrolase release, leading to protoplast and cell wall degradation-typical evidence of programmed cell death (PCD). Therefore, female sterility in the majority of M. obtusifolia sterile ovules is the result of PCD by massive autophagy in the megagametophyte cells. In a few other sterile ovules, sterility is due to the delayed or the absence of megagametophyte development.

Correction to: Ultrastructure and secretion of glandular trichomes in species of subtribe Cajaninae Benth (Leguminosae, Phaseoleae).

In the original version of this article unfortunately some symbols did not appear in the plates caused by technical problems of the journal. We apologize for any inconvenience caused.

Cytological differentiation and cell wall involvement in the growth mechanisms of articulated laticifers in Tabernaemontana catharinensis A.DC. (Apocynaceae).

The cellular mechanisms of laticifer growth are of particular interest in plant biology but are commonly neglected. Using transmission electron microscopy and immunocytochemical methods, we recorded cytological differentiation and evaluated the cell wall involvement in the growth of articulated laticifers with intrusive growth in the mature embryo and plant shoot apex of Tabernaemontana catharinensis. The incorporation of adjacent meristematic cells into the laticifer system occurred in the embryo and plant shoot apex, and the incorporated cells acquired features of laticifer, confirming the laticifers' action-inducing mechanism. In the embryo, this was the main growth mechanism, and began with enlargement of the plasmodesmata and the formation of pores between laticifers and meristematic cells. In the plant shoot apex, it began with loose and disassembled walls and the reorientation of the cortical microtubules of the incorporated cell. Plasmodesmata were absent in these laticifers. There was stronger evidence of intrusive growth in undifferentiated portions of the plant shoot apex than in the embryo. The numerous plasmodesmata in laticifers of the embryo may have been related to the lower frequency of intrusive growth. Intrusive growth was associated with presence of arabinan (increasing wall flexibility and fluidity), and absence of galactan (avoiding wall stiffness), and callose (as a consequence of reduction in symplastic connections) in the laticifer walls. The abundance of low de-methyl-esterified homogalacturonan in the middle lamella and corners may reestablish cell-cell bonding in the laticifers. The cell wall features differed between embryo and plant shoot apex and are directly associated to laticifer growth mechanisms.

Ultrastructure and secretion of glandular trichomes in species of subtribe Cajaninae Benth (Leguminosae, Phaseoleae).

The subtribe Cajaninae of papilionoid legumes has a pantropical distribution and comprises approximately 490 species. These species have diversified throughout dry environments where there are high temperatures and strong light. The subtribe stands out because all its representatives have vesicular glands. In addition, bulbous-based and capitate trichomes are important secretory structures present in all genera of the Cajaninae. We analyzed the ultrastructure and histochemistry of these glandular trichome types in leaflets of the three species of the subtribe. Using transmission electron microscopy and histochemical analyses, we link the glandular secretions to subcellular structures. We here report for the first time the type of exudate and ultrastructure of the glands of subtribe Cajaninae. Terpenoids and phenolics were confirmed by histochemistry tests, and we observed that the organelles responsible for biosynthesis of oils are the most representative in these glands. Each glandular trichome showed particular ultrastructural features compatible with the compounds produced. We suggest that these glandular trichomes, with their respective exudates, act in defense against herbivory and against possible damage by ultraviolet radiation.

Pollen grain development and male sterility in the perfect flowers of Maytenus obtusifolia Mart. (Celastraceae).

Perfect flowers of Maytenus obtusifolia have partial sterility of pollen grains, resulting in collapsed and developed free microspores. However, the cellular events resulting in partial male sterility have not been determined. In pistillate flowers of this species, male sterility is related to the premature programmed cell death (PCD) in tapetum and sporogenic cells. The process occurs through autophagy via macroautophagy and massive autophagy and is associated with sporophytic cytoplasmic male sterility (CMS). Here, we characterised the development of pollen grains and investigated the cellular events that result in tapetum cells and free microspores PCD in perfect flowers, using light and transmission electron microscopy combined with the TUNEL (Terminal deoxynucleotidyl transferase mediated dUDP end-Labeling) assay and the ZIO (Zinc iodide-osmium tetroxide) method. Pollen grain development in perfect flowers was divided into eight developmental stages based on the characteristics of the pollen grains. Tapetum cells undergo PCD at the free microspore stage, through a macroautophagic process, by formation of autophagosomes and by autophagosomes giving rise to lytic vacuoles at maturity. In the final stage of PCD, massive autophagy occurs by rupture of the tonoplast. The development of viable and inviable microspores diverges at the vacuolated microspore stage, when PCD occurs in some free microspores, causing interruption of pollen development through necrosis. These events result in the observed partial male sterility. Viable microspores undergo mitosis and develop into tricellular pollen grains. Male sterility in hermaphrodite individuals is here interpreted as gametophytic CMS.

Structural associations between organelle membranes in nectary parenchyma cells.

MAIN CONCLUSION: The close association between membranes and organelles, and the intense chloroplast remodeling in parenchyma cells of extrafloral nectaries occurred only at the secretion time and suggest a relationship with the nectar secretion. Associations between membranes and organelles have been well documented in different tissues and cells of plants, but poorly explored in secretory cells. Here, we described the close physical juxtaposition between membranes and organelles, mainly with chloroplasts, in parenchyma cells of Citharexylum myrianthum (Verbenaeceae) extrafloral nectaries under transmission electron microscopy, using conventional and microwave fixation. At the time of nectar secretion, nectary parenchyma cells exhibit a multitude of different organelle and membrane associations as mitochondria-mitochondria, mitochondria-endoplasmic reticulum, mitochondria-chloroplast, chloroplast-nuclear envelope, mitochondria-nuclear envelope, chloroplast-plasmalemma, chloroplast-chloroplast, chloroplast-tonoplast, chloroplast-peroxisome, and mitochondria-peroxisome. These associations were visualized as amorphous electron-dense material, a network of dense fibrillar material and/or dense bridges. Chloroplasts exhibited protrusions variable in shape and extension, which bring them closer to each other and to plasmalemma, tonoplast, and nuclear envelope. Parenchyma cells in the pre- and post-secretory stages did not exhibit any association or juxtaposition of membranes and organelles, and chloroplast protrusions were absent. Chloroplasts had peripheral reticulum that was more developed in the secretory stage. We propose that such subcellular phenomena during the time of nectar secretion optimize the movement of signaling molecules and the exchange of metabolites. Our results open new avenues on the potential mechanisms of organelle contact in parenchyma nectary cells, and reveal new attributes of the secretory cells on the subcellular level.

Resin secretory canals in Protium heptaphyllum (Aubl.) Marchand. (Burseraceae): a tridimensional branched and anastomosed system.

Protium heptaphyllum is a Burseraceae species known by the production of aromatic resin with medicinal, economic, and ecological values. Information on the development, architecture, and lifetime of the secretory system are crucial to understand the resin production and contribute to a more sustainable tapping regime. We investigated the histology and ultrastructure of the secretory canals under a developmental point of view. Stem samples were analyzed under light and transmission electron microscopy by conventional and cytochemical methods. Secretory canals, originated from procambium and cambium, occurred immersed in the primary and secondary phloem. Mature canals have a secretory epithelium and a wide lumen where the exudate is accumulated. A sheath of parenchyma cells with meristematic features surrounds the epithelium. The canals originate by schizogenesis and develop by schyzolysigenesis. Canals active in secretion occurred since the shoot apex and near the cambium. In the dilation zone of the secondary phloem, secretory canals exhibit sclerified epithelial and sheath cells and are inactive in secretion. Secreting epithelial cells have subcellular apparatus consistent with oleoresin, polysaccharides, and enzymes secretion. Pectinase and cellulase were cytochemically detected in developing canals and are involved in cell wall changes associated to canal growth and release of exudate. In P. heptaphyllum, the secretory system has a complex structure resultant from longitudinal growth, lateral ramification, and fusion of the adjacent canals, in addition to intrusive growth of both epithelial and sheath cells. Although some anatomical results are already known, ultrastructural data represent the novelty of this work. Our findings can contribute to the establishment of more efficient and sustainable techniques for resin extraction in this species.

Colleters in Rubiaceae from forest and savanna: the link between secretion and environment.

This study aims to investigate colleters' secretory function, on cellular level, in Rubiaceae species from contrasting environments looking to explore the association between secretion and environment. We collected samples from eight species of Rubiaceae growing in forest and savanna having standard-type colleters with diverse histochemistry (hydrophilic, lipophilic and mixed secretions) and processed for both conventional and cytochemical study under transmission electron microscopy (TEM). The standard colleters, although similar in morphology and anatomy, exhibited marked differences on cellular level, especially in the abundance and topology of Golgi bodies, endoplasmic reticulum and plastids when comparing forest and savanna species. These differences were clearly aligned with the chemical nature of the secretions they produce, with predominance of hydrophilic secretions in forest species and lipophilic or mixed secretions in savanna species. The combination of methods in electron microscopy revealed the sites of synthesis and intracellular compartmentation of substances, the mechanisms of their secretion from the protoplast and confirmed the involvement of the outer walls of the epithelial cells in the elimination of exudates to the gland surface. Our study suggests a potential environment-associated plasticity of the secretory cells of standard-type colleters in modulating their secretory function performance.

Structure and functioning of oil cavities in the shoot apex of Metrodorea nigra A. St.-Hil. (Rutaceae).

This study investigates the histology and subcellular features of secretory cavities during the development of the shoot apex of Metrodorea nigra A. St.-Hil. in order to better understand the functioning of these glands. This Rutaceae species is a very suitable model for studying secretory cavity life span, since the shoot apex exhibits both dormant and growth stages during its annual cycle. Shoot apices were collected during the dormant and growth stages from populations of M. nigra growing under natural conditions. Materials were processed using standard techniques for light and electron microscopy. The secretory cavities originate under the protodermis, and their initiation is restricted to the early developmental stage of shoot organs, which are protected by a hood-shaped structure. Secretory cavities have a multi-seriate epithelium surrounding a lumen that expands schizolysigenously. Oil production begins before lumen formation. When the shoot apex resumes development after the dormant stage, the glands remain active in oil secretion in the developing shoot apex and fully expanded leaves. The mature epithelial cells are flattened and exhibit very thin walls, large oil bodies, leucoplasts surrounded by endoplasmic reticulum, and mitochondria with unusual morphology. The tangential walls of the epithelial cells facing the lumen undergo continuous peeling. The vacuole extrusion appears to be the primary mode of release oil into the lumen, in an exocytotic way. The continuity of oil secretion is ensured by the replacement of the damaged inner epithelial cells by divisions in the parenchyma layer that surround the oil gland, likely a meristematic sheath.

Differential bioaccumulation and translocation patterns in three mangrove plants experimentally exposed to iron. Consequences for environmental sensing.

Avicennia schaueriana, Laguncularia racemosa and Rhizophora mangle were experimentally exposed to increasing levels of iron (0, 10, 20 and 100 mg L(-1) added Fe(II) in Hoagland's nutritive medium). The uptake and translocation of iron from roots to stems and leaves, Fe-secretion through salt glands (Avicennia schaueriana and Laguncularia racemosa) as well as anatomical and histochemical changes in plant tissues were evaluated. The main goal of this work was to assess the diverse capacity of these plants to detect mangroves at risk in an area affected by iron pollution (Vitoria, Espírito Santo, Brazil). Results show that plants have differential patterns with respect to bioaccumulation, translocation and secretion of iron through salt glands. L. racemosa showed the best environmental sensing capacity since the bioaccumulation of iron in both Fe-plaque and roots was higher and increased as the amount of added-iron rose. Fewer changes in translocation factors throughout increasing added-iron were observed in this species. Furthermore, the amount of iron secreted through salt glands of L. racemosa was strongly inhibited when exposed to added-iron. Among three studied species, A. schaueriana showed the highest levels of iron in stems and leaves. On the other hand, Rhizophora mangle presented low values of iron in these compartments. Even so, there was a significant drop in the translocation factor between aerial parts with respect to roots, since the bioaccumulation in plaque and roots of R. mangle increased as iron concentration rose. Moreover, rhizophores of R. mangle did not show changes in bioaccumulation throughout the studied concentrations. So far, we propose L. racemosa as the best species for monitoring iron pollution in affected mangroves areas. To our knowledge, this is the first detailed report on the response of these plants to increasing iron concentration under controlled conditions, complementing existing data on the behavior of the same plants under field exposure.