Using disposable food packaging materials as printing, embedding, and sectioning media in the plant anatomy lab.

Premise: During the COVID-19 pandemic lockdown, all laboratory work was suspended, and we were obliged to work from home, causing delays in our research. As the disruption to supply chains made it difficult to obtain our regular lab supplies, we were obliged to search for substitutes. We became familiar with a plastic material known as biaxially oriented polypropylene (BOPP) that is widely used in the food industry for wrapping or storing fruits, vegetables, and meat. BOPP is easily dissolved in organic solvents such as xylenes, acetone, or thinner, but these reagents are very toxic, flammable, and can cause nausea in some users. After testing several alternatives, we found a polyurethane remover that proved to be an effective and relatively harmless BOPP solvent. Methods and Results: By dissolving thin slices of BOPP in a polyurethane solvent, we obtained a clean fluid that we used to obtain leaf surface prints that could be mounted on microscope slides with a coverslip. This fluid produced excellent bark and wood sections and can be used to obtain wood or charcoal surface prints. Our attempts to use it as a mounting medium were unsuccessful. Conclusions: BOPP dissolved in a polyurethane remover is a handy, versatile resource for plant microtechniques. In addition to its economic advantages, it is useful in terms of reducing plastic pollution.

Premisa: Durante el cierre por pandemia de COVID­19, se suspendió todo el trabajo de laboratorio y nos vimos obligados a trabajar desde casa, lo que provocó retrasos en nuestras investigaciones. Como la interrupción de las cadenas de suministro dificultó la obtención de nuestros suministros de laboratorio habituales, nos vimos obligados a buscar sustitutos. Nos familiarizamos con un material plástico empleado en la industria alimentaria, muy utilizado para envolver o almacenar frutas, verduras y carne. Este material se conoce como polipropileno orientado biaxialmente (BOPP, en inglés) y se disuelve fácilmente en disolventes orgánicos como xilenos, acetona o tíner. Sin embargo, estos reactivos son muy tóxicos e inflamables y pueden provocar náuseas a algunos usuarios. Tras probar varias alternativas, encontramos un removedor de poliuretano que demostró ser un disolvente eficaz para el BOPP. Métodos y Resultados: Disolviendo tiras delgadas de BOPP en un disolvente de poliuretano, obtuvimos un fluido limpio que utilizamos para obtener impresiones de la superficie de las hojas que podían montarse en portaobjetos de microscopio con un cubreobjetos. Este fluido produce excelentes secciones de corteza y madera y se puede usar para obtener impresiones de superficies de madera o carbón. Intentamos utilizarlo como medio de montaje, pero desistimos debido a los pobres resultados obtenidos. Conclusión: El BOPP disuelto en removedor de poliuretano es un recurso práctico y versátil para las microtécnicas vegetales. Además de sus ventajas económicas, es útil para evitar que el plástico llegue a las corrientes de agua y a los océanos.

Shifts in soil and plant functional diversity along an altitudinal gradient in the French Alps.

OBJECTIVES: Altitude integrates changes in environmental conditions that determine shifts in vegetation, including temperature, precipitation, solar radiation and edaphogenetic processes. In turn, vegetation alters soil biophysical properties through litter input, root growth, microbial and macrofaunal interactions. The belowground traits of plant communities modify soil processes in different ways, but it is not known how root traits influence soil biota at the community level. We collected data to investigate how elevation affects belowground community traits and soil microbial and faunal communities. This dataset comprises data from a temperate climate in France and a twin study was performed in a tropical zone in Mexico. DATA DESCRIPTION: The paper describes soil physical and chemical properties, climatic variables, plant community composition and species abundance, plant community traits, soil microbial functional diversity and macrofaunal abundance and diversity. Data are provided for six elevations (1400-2400 m) ranging from montane forest to alpine prairie. We focused on soil biophysical properties beneath three dominant plant species that structure local vegetation. These data are useful for understanding how shifts in vegetation communities affect belowground processes, such as water infiltration, soil aggregation and carbon storage. Data will also help researchers understand how plant communities adjust to a changing climate/environment.

A novel, rapid technique for clearing leaf tissues.

PREMISE: Clearing leaves is a highly useful practice for many taxonomic, ecological, physiological, and eco-physiological aspects of research. Using traditional methods, the procedure for clearing a leaf (referred to as diaphanization) can take several days or even weeks. In our laboratory we developed a technique, originally used for dissociating wood, that yields excellent epidermal and leaf venation preparations clearly showing the details of epidermal cells, hydathodes, trichomes, leaf margins, and leaf venation, in a maximum of three days. METHODS AND RESULTS: A solution originally used to macerate wood (Franklin's solution) was used to accelerate the process of clearing leaves. Using this approach, it is possible to obtain clear images of the leaf adaxial and abaxial epidermal surfaces and leaf venation. Our technique works with both fragile and sturdy leaves, as well as with thin roots and stems. A few examples are presented. CONCLUSIONS: Franklin's solution is very simple to prepare and easy to manipulate. This new technique has the advantage of separating the leaf epidermal layers and venations, which can then be clearly observed.

The ecophysiology of a neotropical mistletoe depends on the leaf phenology of its tree hosts.

PREMISE: Mistletoes parasitize many hardwood and softwood tree species; however, they play key roles in forest ecosystems. Adult individuals of Psittacanthus schiedeanus take up water and xylem nutrients from both deciduous and evergreen host trees, suggesting the ability to modify its physiology according to the availability of host resources. Yet, there is little information regarding the effects of mistletoes on their host trees from the eophyll stage to reproductive phases of the parasite. METHODS: Taking advantage of the fact that P. schiedeanus can reach sexual maturity in 1 year, we investigated its physiological performance during development on deciduous (Liquidambar styraciflua) and evergreen (Quercus germana) host trees in a cloud forest in eastern Mexico. Variables related to chlorophyll fluorescence, carbon assimilation, photosynthetic pigments, and nitrogen, phosphorus, and carbon contents of the parasite and non-infected and infected hosts were analyzed in a nursery experiment. RESULTS: Mistletoe had lower water-use efficiency and higher transpiration rates than the host species did. Despite the fact that P. schiedeanus obtained resources from species with differing phenology and resource availability, the parasite steadily improved its CO2 assimilation, electron transport rate, and nutrient content from seedling establishment to adult life stages. Mistletoe decreased the photosynthetic reactions of carbon metabolism in the deciduous host, photosynthetic light reactions in the evergreen host, and nutritional status of both host species, mostly in the evergreen host. CONCLUSIONS: The hypothesis that mistletoes adjust their physiology according to the availability of host resources could extend to the early growth of the parasite.

Novel findings to the biosynthetic pathway of magnoflorine and taspine through transcriptomic and metabolomic analysis of Croton draco (Euphorbiaceae).

BACKGROUND: Croton draco is an arboreal species and its latex as well as some other parts of the plant, are traditionally used in the treatment of a wide range of ailments and diseases. Alkaloids, such as magnoflorine, prevent early atherosclerosis progression while taspine, an abundant constituent of latex, has been described as a wound-healer and antitumor-agent. Despite the great interest for these and other secondary metabolites, no omics resources existed for the species and the biosynthetic pathways of these alkaloids remain largely unknown. RESULTS: To gain insights into the pathways involved in magnoflorine and taspine biosynthesis by C. draco and identify the key enzymes in these processes, we performed an integrated analysis of the transcriptome and metabolome in the major organs (roots, stem, leaves, inflorescences, and flowers) of this species. Transcript profiles were generated through high-throughput RNA-sequencing analysis while targeted and high resolution untargeted metabolomic profiling was also performed. The biosynthesis of these compounds appears to occur in the plant organs examined, but intermediaries may be translocated from the cells in which they are produced to other cells in which they accumulate. CONCLUSIONS: Our results provide a framework to better understand magnoflorine and taspine biosynthesis in C. draco. In addition, we demonstrate the potential of multi-omics approaches to identify candidate genes involved in the biosynthetic pathways of interest.

Effect of Resin Ducts and Sap Content on Infestation and Development of Immature Stages of Anastrepha obliqua and Anastrepha ludens (Diptera: Tephritidae) in Four Mango (Sapindales: Anacardiaceae) Cultivars.

We determined the influence of resin ducts, sap content, and fruit physicochemical features of four mango cultivars (Criollo, Manila, Ataulfo, and Tommy Atkins) on their susceptibility to the attack of the two most pestiferous fruit fly species infesting mangoes in Mexico: Anastrepha ludens (Loew) and Anastrepha obliqua (Macquart). We performed three studies: 1) analysis of resin ducts in mango fruit exocarp to determine the density and area occupied by resin ducts in each mango cultivar, 2) assessment of mango physicochemical features including fruit sap content, and 3) a forced infestation trial under field conditions using enclosed fruit-bearing branches to expose mangoes to gravid A. ludens or A. obliqua females. Infestation rates, development time from egg to prepupae and pupae, pupal weight, and percent of adult emergence, were assessed. 'Ataulfo' and 'Tommy Atkins' cultivars exhibited the highest resin duct density and sap content, the lowest infestation rate, and had a negative effect on immature development and pupal weight. In sharp contrast, 'Manila' and 'Criollo' cultivars, with the lowest resin duct density and sap content, were highly susceptible to A. ludens and A. obliqua attack. We conclude that sap content and the number, size, and distribution of resin ducts as well as firmness in mango fruit exocarp are all involved in the resistance of mango to A. ludens and A. obliqua attack.

Bidirectional anatomical effects in a mistletoe-host relationship: Psittacanthus schiedeanus mistletoe and its hosts Liquidambar styraciflua and Quercus germana.

PREMISE OF THE STUDY: During the interactions between a parasitic plant and its host, the parasite affects its host morphologically, anatomically, and physiologically, yet there has been little focus on the effect of hosts on the parasite. Here, the functional interactions between the hemiparasitic mistletoe Psittacanthus schiedeanus and its hosts Liquidambar styraciflua and Quercus germana were interpreted based on the anatomical features of the vascular tissues. METHODS: Using standard techniques for light and transmission electron microscopy, we studied the effects of P. schiedeanus on the phloem anatomy of Liquidambar styraciflua and Quercus germana and vice versa. KEY RESULTS: The phloem of P. schiedeanus has larger sieve elements, companion cells, and sieve plate areas when it is parasitizing L. styraciflua than Q. germana; however, the parasite produces systemic effects on the phloem of its hosts, reducing the size of phloem in L. styraciflua but increasing it in Q. germana. Those seem to be the bidirectional effects. No direct connections between the secondary phloem of the parasite and that of its hosts were observed. Parenchymatic cells of L. styraciflua in contact with connective parenchyma cells of the parasite develop half-plasmodesmata, while those of Q. germana do not. CONCLUSIONS: The bidirectional effects between the parasite and its hosts comprise modifications in secondary phloem that are potentially affected by the phenology of its hosts, a combination of hormonal agents such as auxins, and the symplasmic or apoplasmic pathway for solutes import.

Structure and histology of extrafloral nectaries of tropical species in a mexican rain forest / Estrutura e histologia dos nectarios extraflorais de especies em uma floresta tropical no México

Although there is a large diversity of plant species with extrafloral nectaries, histological detail of these glands is poorly documented in tropical rain forest species. We characterized extrafloral nectaries using digital photographs, scanning electron microscopy and histological methods for eight plant species belonging to the Costaceae, Euphorbiaceae, Malvaceae and Salicaceae. To our knowledge, there are no previous reports on the structure of extrafloral nectaries for these species. Croton species exhibited reddish extrafloral nectaries on the petioles, surrounded by starshaped trichomes. Cnidoscolus multilobus and Omphalea oleifera showed green, elevated, extrafloral nectaries at the peduncle; C. multilobus showed abundant secretory tissue; O. oleifera revealed numerous cells with calcium oxalate crystals. Heliocarpus species showed extrafloral nectaries distributed at the base of the blade and possesses glands in the center of the extrafloral nectary surrounded by a series of parenchymatic cells. Pleuranthodendron lindenii has two extrafloral nectaries at the base of the leaves. Costus scaber has a hollow-type extrafloral nectary on the red-colored bracts of the inflorescence; nectar cavity is found in the center area of each bracts located on a yellow line. The extrafloral nectaries described in this study exhibited different morphologies and histological structures involved in the secretion of extrafloral nectar that could be related to biotic defenses, primarily by attracting ants.

Embora exista uma grande diversidade de espécies de plantas com nectários extraflorais, detalhes histológicos destas glândulas ainda é pouco documentado para espécies tropicais. Nós caracterizamos nectários extraflorais usando fotografias digitais, microscopia eletrônica de varredura e métodos histológicos para oito espécies de plantas das famílias Costaceae, Euphorbiaceae, Malvaceae e Salicaceae. Para nosso conhecimento, não há relatos anteriores sobre a estrutura dos nectários extraflorais para as espécies estudadas. Espécies de Croton exibiram nectários extraflorais avermelhadas nos pecíolos, rodeados por tricomas em forma de estrela. Cnidoscolus multilobus e Omphalea oleifera apresentaram nectários extraflorais verdes e elevados nos pedúnculos; C. multilobus apresenta tecido secretor abundante enquanto que O. oleifera apresentou numerosas células com cristais de oxalato de cálcio. Espécies de Heliocarpus apresentaram nectários extraflorais distribuídos na base da lâmina e possuíam glândulas no centro do nectário rodeadas por uma série de células de parênquima. Pleuranthodendron lindenii apresentou dois nectários extraflorais na base das folhas. Costus scaber apresenta um nectário extrafloral do tipo oco nas brácteas avermelhadas da inflorescência, e a cavidade do nectário se encontra na área central de cada bráctea localizados em uma linha amarela. Os nectários extraflorais descritos neste estudo apresentaram morfologia e estruturas histológicas diferentes envolvidas na secreção de néctar extrafloral que poderia estar relacionado com defesas bióticas, principalmente atraindo formigas.

Exploiting water versus tolerating drought: water-use strategies of trees in a secondary successional tropical dry forest.

In seasonal plant communities where water availability changes dramatically both between and within seasons, understanding the mechanisms that enable plants to exploit water pulses and to survive drought periods is crucial. By measuring rates of physiological processes, we examined the trade-off between water exploitation and drought tolerance among seedlings of trees of a tropical dry forest, and identified biophysical traits most closely associated with plant water-use strategies. We also explored whether early and late secondary successional species occupy different portions of trade-off axes. As predicted, species that maintained carbon capture, hydraulic function and leaf area at higher plant water deficits during drought had low photosynthetic rates, xylem hydraulic conductivity and growth rate under non-limiting water supply. Drought tolerance was associated with more dense leaf, stem and root tissues, whereas rapid resource acquisition was associated with greater stem water storage, larger vessel diameter and larger leaf area per mass invested. We offer evidence that the water exploitation versus drought tolerance trade-off drives species differentiation in the ability of tropical dry forest trees to deal with alternating water-drought pulses. However, we detected no evidence of strong functional differentiation between early and late successional species along the proposed trade-off axes, suggesting that the environmental gradient of water availability across secondary successional habitats in the dry tropics does not filter out physiological strategies of water use among species, at least at the seedling stage.

Root biomechanics in Rhizophora mangle: anatomy, morphology and ecology of mangrove's flying buttresses.

BACKGROUND AND AIMS: Rhizophora species of mangroves have a conspicuous system of stilt-like roots (rhizophores) that grow from the main stem and resemble flying buttresses. As such, the development of rhizophores can be predicted to be important for the effective transmission of dynamic loads from the top of the tree to the ground, especially where the substrate is unstable, as is often the case in the habitats where Rhizophora species typically grow. This study tests the hypothesis that rhizophore architecture in R. mangle co-varies with their proximity to the main stem, and with stem size and crown position. METHODS: The allometry and wood mechanical properties of R. mangle (red mangrove) trees growing in a mangrove basin forest within a coastal lagoon in Mexico were compared with those of coexisting, non-buttressed mangrove trees of Avicennia germinans. The anatomy of rhizophores was related to mechanical stress due to crown orientation (static load) and to prevailing winds (dynamic load) at the study site. KEY RESULTS: Rhizophores buttressed between 10 and 33 % of tree height. There were significant and direct scaling relationships between the number, height and length of rhizophores vs. basal area, tree height and crown area. Wood mechanical resistance was significantly higher in the buttressed R. mangle (modulus of elasticity, MOE = 18·1 ± 2 GPa) than in A. germinans (MOE = 12·1 ± 0·5 GPa). Slenderness ratios (total height/stem diameter) were higher in R. mangle, but there were no interspecies differences in critical buckling height. When in proximity to the main stem, rhizophores had a lower length/height ratio, higher eccentricity and higher xylem/bark and pith proportions. However, there were no directional trends with regard to prevailing winds or tree leaning. CONCLUSIONS: In comparison with A. germinans, a tree species with wide girth and flare at the base, R. mangle supports a thinner stem of higher mechanical resistance that is stabilized by rhizophores resembling flying buttresses. This provides a unique strategy to increase tree slenderness and height in the typically unstable substrate on which the trees grow, at a site that is subject to frequent storms.

Dynamic control of osmolality and ionic composition of the xylem sap in two mangrove species.

• Premise of the study: Xylem sap osmolality and salinity is a critical unresolved issue in plant function with impacts on transport efficiency, pressure gradients, and living cell turgor pressure, especially for halophytes such as mangrove trees.• Methods: We collected successive xylem vessel sap samples from stems and shoots of Avicennia germinans and Laguncularia racemosa using vacuum and pressure extraction and measured their osmolality. Following a series of extractions with the pressure chamber, we depressurized the shoot and pressurized again after various equilibration periods (minutes to hours) to test for dynamic control of osmolality. Transpiration and final sap osmolality were measured in shoots perfused with deionized water or different seawater dilutions.• Key results: For both species, the sap osmolality values of consecutive samples collected by vacuum extraction were stable and matched those of the initial samples extracted with the pressure chamber. Further extraction of samples with the pressure chamber decreased sap osmolality, suggesting reverse osmosis occurred. However, sap osmolalities increased when longer equilibration periods after sap extraction were allowed. Analysis of expressed sap with HPLC indicated a 1:1 relation between measured osmolality and the osmolality of the inorganic ions in the sap (mainly Na+, K+, and Cl-), suggesting no contamination by organic compounds. In stems perfused with deionized water, the sap osmolality increased to mimic the native sap osmolality.• Conclusions: Xylem sap osmolality and ionic contents are dynamically adjusted by mangroves and may help modulate turgor pressure, hydraulic conductivity, and water potential, thus being important for mangrove physiology, survival, and distribution.

Anatomical and physical changes in leaves during the production of tamales.

PREMISE OF THE STUDY: Tamale preparation has a long tradition in Mexico. To understand which material properties have been considered important for this purpose throughout the years, a study was conducted of the anatomical, chemical, and mechanical properties of the leaves of four plant species used in tamale preparation in Veracruz, Mexico: Calathea misantlensis, Canna indica, Musa paradisiaca, and Oreopanax capitatus. METHODS: Four cooking treatments were considered: fresh (F), roasted (soasado, R), steamed (S), and roasted plus steamed (R/S). Chemical, anatomical, and mechanical analyses were conducted before and after each treatment. Leaf samples were tested for tensile strength at both parallel and perpendicular orientation relative to the fibers. KEY RESULTS: Musa paradisiaca had the highest proportion of cellulose, while the remaining species shared similar lower proportions. Leaves were stronger and stiffer in the longitudinal direction of the fibers. Musa paradisiaca leaves had higher values of mechanical strength than the other species. The cooking process that most affected the mechanical properties was steaming. CONCLUSIONS: The chemical constituents of the leaves are closely correlated with their physical properties. The treatment that caused the greatest decrease in leaf physical integrity was steaming, while the combination of roasting and steaming showed similar results to those of steaming alone. No evident anatomical changes are produced by any of the treatments. This is one of the few studies comparing physical, chemical, and anatomical characteristics of leaves used for human consumption, before and after cooking.

Bark anatomy in Croton draco var. draco (Euphorbiaceae).

Many arboreal forms of the genus Croton (ca. 800 spp.), amply distributed in the Americas, have latex-producing cells in their bark, which is widely used in traditional medicine to treat skin infections and some forms of cancer. Studies validate its ethnomedicinal use-more than 20 pharmaceutically important secondary metabolites have been reported for its latex and bark-but anatomical and ecological studies are scarce. Given this species' ample distribution, laticifer abundance could be affected by the environment. We tested this for genetically similar trees growing in two types of vegetation in Veracruz, Mexico at sites commonly visited by traditional doctors. We describe the bark anatomy of C. draco, focusing on the laticifers, histochemically characterize the bark and the latex extracted from it, and document differences in laticifer abundance in the two environments. We have also identified another cell type (what we call type B) in the secretory latex system and describe it histochemically and microscopically. The location of bark cells that contain essential oils is reported here for the first time. Given the genetic similarity of the trees at both sites, the between-site variation in the number of laticifers in stem and branch bark appears to be an effect of the environment.

Anatomía de seis especies de helechos del género Dryopteris (Dryopteridaceae) de México / Anatomy of six species of Dryopteris ferns (Dryopteridaceae) from Mexico

Rhizome and foliar anatomy of the Mexican Dryopteris Adans. species were studied and compared with other Dryopteridaceae and other fern families to identify anatomical features with diagnostic value. The anatomy of rhizome, stipe, and blade is similar in species of the Dryopteris patula complex. The cells with un-lignified, thickened wall, with cap or U-shape around the meristeles belong to the collenchyma, in contrast with other fern families. Dryopteris wallichiana (Spreng.) Hyl. is anatomically distinguished from the other studied species by having more layers of sclerenchyma and meristeles on the stipe, and by the lack of sclereid nests on the rhizome. Dryopteris rossii C. Chr. and D. maxonii Underw. & C. Chr. are characterized by the presence of crystals on the periphery of rhizome nests. D. maxonni and D. wallichiana lack blade glands.

Se estudió la anatomía del rizoma y hoja de especies mexicanas de Dryopteris Adans. y se comparó con la información disponible para Dryopteridaceae y otras familias de helechos en busca de caracteres anatómicos con valor diagnóstico. La anatomía de rizoma, pecíolo y lámina es similar en las especies del complejo Dryopteris patula. Las células con pared engrosada, no lignificada y en forma de casquete o de “U” presentes alrededor de las meristelas corresponden a colénquima, a diferencia de lo informado para otras familias de helechos. Dryopteriswallichiana (Spreng.) Hyl. se distingue anatómicamente de las otras especies estudiadas por presentar un mayor número de capas de esclerénquima y de meristelas en el pecíolo, además de carecer de nidos de esclereidas en el rizoma. Dryopteris rossii y D. maxonii se caracterizan por la presencia de cristales en la periferia de los nidos en el rizoma. Las glándulas en la lámina están ausentes en D. maxonii y D. wallichiana.

Preparative laser capture microdissection and single-pot cell wall material preparation: a novel method for tissue-specific analysis.

In adaptation to their function the walls of plant cell display tissue-specific variations of composition according to their developmental stage, cell type and stress of various origin. It is therefore important to obtain a precise analytical data describing the cell wall composition with respect to these different factors. In the present work, laser capture microdissection (LCM) was used for isolating different tissues from the stem of Urtica dioica L. at a semi-preparative scale. The technique was associated for the first time to a one-pot sequential cell wall preparation and hydrolysis for the carbohydrate analysis of each cell type. The results demonstrate that the combination of LCM and micro-analytical methods can provide individual cell type composition and should improve our knowledge of the biochemical diversity of cell walls in plants. This approach will be of potential interest for the understanding of the effects of stress or genetic engineering on the composition of the cell walls.

[Anatomy of six species of Dryopteris ferns (Dryopteridaceae) from Mexico]. / Anatomía de seis especies de helechos del género Dryopteris (Dryopteridaceae) de México.

Rhizome and foliar anatomy of the Mexican Dryopteris Adans. species were studied and compared with other Dryopteridaceae and other fern families to identify anatomical features with diagnostic value. The anatomy of rhizome, stipe, and blade is similar in species of the Dryopteris patula complex. The cells with un-lignified, thickened wall, with cap or U-shape around the meristeles belong to the collenchyma, in contrast with other fern families. Dryopteris wallichiana (Spreng.) Hyl. is anatomically distinguished from the other studied species by having more layers of sclerenchyma and meristeles on the stipe, and by the lack of sclereid nests on the rhizome. Dryopteris rossii C. Chr. and D. maxonii Underw. & C. Chr. are characterized by the presence of crystals on the periphery of rhizome nests. D. maxonni and D. wallichiana lack blade glands.

Morphological and secretory characterization of extrafloral nectaries in plants of coastal Veracruz, Mexico.

BACKGROUND AND AIMS: Morphological descriptions of the extrafloral nectaries (EFNs) of certain plant species are common in the literature, but they rarely relate morphology with histology, gland distribution and secretory attributes. In this study a morphological/secretory characterization of EFNs occurring on several plant species in a tropical coastal community is made and the implications of gland attributes discussed from a functional perspective. METHODS: The morphology and nectar secretion of the EFNs of 20 plant species are characterized through scanning electron microscopy, histochemical detection of reducing sugars (Fehling's reagent) and nectar volume/concentration estimates. KEY RESULTS: Sixty-five per cent of plant species in coastal communities had EFNs on vegetative structures and 35 % of species had glands on reproductive and vegetative organs. The Fabaceae is the plant family with the most species with EFNs and most diversity of gland morphologies. Four types of vascularized nectaries and four of glandular trichomes are described; sugar-secreting trichomes are characterized using Fehling's technique, and the first descriptions of unicellular and peltate trichomes functioning as EFNs are provided. Glands of ten plant species and six genera are described for the first time. Four plant species possess more than one morphological type of EFN. Eleven species have EFNs in more than one location or organ. More complex glands secrete more nectar, but are functionally homologous to the aggregations of numerous secretory trichomes on specific and valuable plant organs. CONCLUSION: Important diversity of EFN morphology was foundin the coastal plant community studied. Both vascularized and non-vascularized EFNs are observed in plants and, for the latter, previously non-existent morpho-secretory characterizations are provided with a methodological approach to study them. It is recommended that studies relating EFN attributes (i.e. morphology, distribution) with their differential visitation by insects (i.e. ants) and the cost of maintenance to the plants are carried out to understand the evolution of these glands.

Hydraulic conductivity and embolism in the mangrove tree Laguncularia racemosa.

We measured xylem pressure potentials, soil osmotic potentials, hydraulic conductivity and percent loss of conductivity (PLC) due to embolism, and made microscopic observations of perfused dye in the white mangrove tree, Laguncularia racemosa (L.) Gaertn. f., (1) to determine its vulnerability to air embolism compared with published results for the highly salt-tolerant red mangrove tree, Rhizophora mangle L., and (2) to identify possible relationships between air embolism, permanent blockage of vessels and stem diameter. Laguncularia racemosa was more vulnerable to embolism than reported for R. mangle, with 50 PLC at -3.4 MPa. Narrow stems (5-mm diameter) had higher PLC than larger stems (8.4- or 14-mm diameter) of the same plants. Basic fuchsin dye indicated that up to 89% of the vessels, especially in the narrow stems, had permanent blockage that could not be reversed by high pressure perfusion. Air embolism could lead to permanent vessel blockage and eventual stem mortality. Such vulnerability to embolism may restrict the growth of L. racemosa and limit its distribution to less salty areas of mangrove communities.

Fluorescence shell: a novel view of sclereid morphology with the Confocal Laser Scanning Microscope.

Confocal Laser Scanning Microscopy (CLSM) was used to observe sclereids from stems of Avicennia germinans and from fruits of two species of pear (Pyrus calleryana "Bradford" and P. communis "Red Bartlett"). The images obtained from thick (25 to 100 microm) free-hand sections were, in certain respects, far superior to those obtained by other, more invasive and time-consuming microscopic techniques upon which previous reports of sclereid morphology were based. The cell wall surfaces, including the "internal" surfaces of the branched pit canals and cell lumens, were much accentuated with the techniques we describe, resulting in a "fluorescence shell" image, meaning the cell wall did not stain all the way through but instead only at the inner and outer wall surfaces, including the edges of ramiform pits. By controlling the time of staining with 1% aqueous Safranin O, or by changing the number of optical sections used in extended focus images, it was possible to get either a conventional view of the cell wall structure or a unique, three-dimensional view of the elaborate cell interconnections. Similar fluorescence shell images of sclereids were also obtained using a periodic-Schiff (PAS) staining system, but the stain was not as specific to sclereid cell walls as was the Safranin O stain. Particularly with the use of a narrow range band pass emission filter of 505-530 nm, the Safranin O staining may be more specific to lignin than reported in the literature.