Stream food webs in tropical mountains rely on allochthonous carbon regardless of land use.

The relative importance of allochthonous and autochthonous carbon (C) as sources of energy for tropical stream food webs remains an open question. Allochthonous C might be the main energy source for small and shaded forest streams, while autochthonous C is more likely to fuel food webs draining land uses with less dense vegetation. We studied food webs in cloud forest streams draining watersheds with forests, coffee plantations, and pastures. Our goal was to assess the effects of those land uses on the C source and structure of stream food webs. The study took place in tropical montane streams in La Antigua Watershed, in eastern Mexico. We selected three streams per land use and sampled biofilm and leaf litter as the main food resources, and macroinvertebrates and aquatic vertebrates from different trophic guilds. Samples were analyzed for δ13C and δ15N isotopes. Using a Bayesian mixing model, we estimated the proportional assimilation of autochthonous and allochthonous carbon by each guild. We found that consumers were mostly using allochthonous C in all streams, regardless of watershed land use. Our findings indicate that montane cloud forest streams are dominated by allochthony even in watersheds dominated by pastures. Abundant precipitation in this life zone might facilitate the movement of allochthonous C into streams. While food webs of streams from coffee plantations and pastures also rely on allochthonous resources, other impacts do result in important changes in stream functioning.

Non-chlorophyllous and crypto-chlorophyllous fern spores differ in their mobilisation of fatty acids during priming.

During fern spore germination, lipid hydrolysis primarily provides the energy to activate their metabolism. In this research, fatty acids (linoleic, oleic, palmitic and stearic) were quantified in the spores exposed or not to priming (hydration-dehydration treatments). Five fern species were investigated, two from xerophilous shrubland and three from a cloud forest. We hypothesised that during the priming hydration phase, the fatty acids profile would change in concentration, depending on the spore type (non-chlorophyllous and crypto-chlorophyllous). The fatty acid concentration was determined by gas chromatograph-mass spectrometer. Chlorophyll in spores was vizualised by epifluorescence microscopy and quantified by high-resolution liquid chromatography with a DAD-UV/Vis detector. Considering all five species and all the treatments, the oleic acid was the most catabolised. After priming, we identified two patterns in the fatty acid metabolism: (1) in non-chlorophyllous species, oleic, palmitic, and linoleic acids were catabolised during imbibition and (2) in crypto-chlorophyllous species, these fatty acids increased in concentration. These patterns suggest that crypto-chlorophyllous spores with homoiochlorophylly (chlorophyll retained after drying) might not require the assembly of new photosynthetic apparatus during dark imbibition. Thus, these spores might require less energy from pre-existing lipids and less fatty acids as 'building blocks' for cell membranes than non-chlorophyllous spores, which require de novo synthesis and structuring of the photosynthetic apparatus.

A reduced role for water transport during the Cenozoic evolution of epiphytic Eupolypod ferns.

The Cretaceous-Cenozoic expansion of tropical forests created canopy space that was subsequently occupied by diverse epiphytic communities including Eupolypod ferns. Eupolypods proliferated in this more stressful niche, where lower competition enabled the adaptive radiation of thousands of species. Here, we examine whether xylem traits helped shape the Cenozoic radiation of Eupolypod ferns. We characterized the petiole xylem anatomy of 39 species belonging to the Eupolypod I and Eupolypod II clades occupying the epiphytic, hemiepiphytic, and terrestrial niche, and we assessed vulnerability to embolism in a subset of species. The transition to the canopy was associated with reduced xylem content and smaller tracheid diameters, but no differences were found in species vulnerability to embolism and pit membrane thickness. Phylogenetic analyses support selection for traits associated with reduced water transport in Eupolypod 1 species. We posit that in Eupolypod epiphytes, selection favored water retention via thicker leaves and lower stomatal density over higher rates of water transport. Consequently, lower leaf water loss was coupled with smaller quantities of xylem and narrower tracheid diameters. Traits associated with water conservation were evident in terrestrial Eupolypod 1 ferns and may have predisposed this clade toward radiation in the canopy.

Terminal velocity of fern and lycopod spores is affected more by mass and ornamentation than by size.

PREMISE: Terminal velocity (Vt) is an important factor for the dispersal of biological particles but has scarcely been studied for anemochorous fern spores, and the influence of spore characteristics on Vt has not been evaluated. Here, we measured the Vt of 1234 spores of 18 fern species and two Selaginella microspores using videoimaging analysis and evaluated the effects of mass, size, and ornamentation on Vt. METHODS: We designed a sedimentation tower with a graduated microtelescope attached to a high-speed video camera to record falling particles and measure the Vt of fern spores using video-image processing software. Spores were measured for each species and their size correlated with Vt. RESULTS: The Vt of fern spores ranged from 4.7 cm·s-1 (Cyathea costaricensis) to 18.85 cm·s-1 (Acrostichum danaeifolium). The method is accurate and reliable as predicted by Stokes model for glass beads of known density and size. In addition, Vt had a higher correlation coefficient with mass (ρ = 0.72) than size (ρ = 0.20), and ornamental appendages reduced Vt. CONCLUSIONS: The reported values of Vt of fern spores are within the range of different biological airborne particles such as moss spores and pollen grains of seed plants. The results showed that spore ornamentation is directly related to Vt rather than spore size and may increase or decrease the drag. This method will aid future aerobiological research on biological particles.

Hydathodes in ferns: their phylogenetic distribution, structure and function.

BACKGROUND AND AIMS: Ferns are the second largest group of vascular plants and are distributed nearly worldwide. Although ferns have been integrated into some comparative ecological studies focusing on hydathodes, there is a considerable gap in our understanding of the functional anatomy of these secretory tissues that are found on the vein endings of many fern leaves. In this study, we aimed to investigate the phylogenetic distribution, structure and function of fern hydathodes. METHODS: We performed a global review on fern hydathodes and their phylogenetic distribution, carried out an ancestral character state reconstruction, and studied the structure, guttation and elemental composition of salt residues of eight species, and the diurnal patterns of xylem pressure of two species. KEY RESULTS: Hydathodes are known from 1189 fern species, 92 genera and 19 families of 2 orders, Equisetales and Polypodiales. Stochastic character mapping indicated multiple gains and losses of hydathodes at the genus level, occurring especially during the last 50 million years of fern evolution. Hydathodes were located on the adaxial leaf surface and characterized by a cytoplasm-rich, pore-free epidermis, and became functional for several weeks after nearly complete leaf expansion. In two species, positive xylem pressure built up at night, potentially facilitating guttation. Guttation fluid was rich in Ca and often Si, but also contained P, Mg, Na and Al. CONCLUSIONS: Stochastic character mapping and the structural and functional diversity of hydathodes indicate multiple origins, and their presence/absence in closely related taxa implies secondary losses during fern evolution. Positive xylem pressure and high air humidity play an important role as drivers of guttation. Hydathodes may contribute to the regulation of leaf nutrient stoichiometry by the release of excessive compounds and minerals other than waste products, but the presence of essential chemical elements in salt residues also indicates possible leakage.

Viability markers for determination of desiccation tolerance and critical stages during dehydration in Selaginella species.

While most plants die below a threshold of water content, desiccation-tolerant species display specific responses that allow them to survive extreme dehydration. Some of these responses are activated at critical stages during water loss and could represent the difference between desiccation tolerance (DT) and death. Here, we report the development of a simple and reproducible system to determine DT in Selaginella species. The system is based on exposure of excised tissue to a dehydration agent inside small containers, and subsequent evaluation for tissue viability. We evaluated several methodologies to determine viability upon desiccation including: triphenyltetrazolium chloride (TTC) staining, the quantum efficiency of PSII, antioxidant potential, and relative electrolyte leakage. Our results show that the TTC test is a simple and accurate assay to identify novel desiccation-tolerant Selaginella species, and can also indicate viability in other desiccation-tolerant models (i.e. ferns and mosses). The system we developed is particularly useful to identify critical points during the dehydration process. We found that a desiccation-sensitive Selaginella species shows a change in viability when dehydrated to 40% relative water content, indicating the onset of a critical condition at this water content. Comparative studies at critical stages could provide a better understanding of DT mechanisms and unravel insights into the key responses to survive desiccation.

Nectaries in ferns: their taxonomic distribution, structure, function, and sugar composition.

PREMISE: Extrafloral nectaries have mainly been studied in angiosperms, but have also been reported in 39 fern species. Here we provide a global review of nectaries in ferns and examined their structure, function, and nectar sugar composition in two genera. METHODS: We searched in the literature and living plant collections of botanical gardens for indications of fern nectaries, observed nectar-feeding animals, studied the morphoanatomy in the two genera Aglaomorpha and Campyloneurum, and analyzed the total sugar concentrations and ratios of 16 species. Diurnal nectar release was observed with time-lapse photography. RESULTS: We found evidence for nectaries in 101 species of ferns from 11 genera and 6 families. Most of the nectary-bearing species were tree ferns (Cyatheaceae) and epiphytic ferns of the family Polypodiaceae. Nectaries consisted of cytoplasm-rich parenchyma with large nuclei and an epidermis with or without stomata, were attached to amphiphloic vascular bundles, and released nectar on the lower leaf surface mainly on expanding leaves during the night. Sugar concentrations varied between species (3.8-15.3%) but not between genera, and were sucrose-dominant (3 spp.), sucrose-rich (7), or hexose-rich (3). In the greenhouse, introduced ants, scale insects, and snails fed on the nectar. CONCLUSIONS: The wide taxonomic distribution, variable morphology, locations, and sugar compositions point to multiple evolutionary origins of fern nectaries. Nectar release in young leaves might attract mutualistic ants to protect leaves against herbivores only during this most vulnerable developmental stage. Even ex-situ, fern nectar is a valuable food source because it attracted several opportunistic animal species.

Leaf water relations in epiphytic ferns are driven by drought avoidance rather than tolerance mechanisms.

Opportunistic diversification has allowed ferns to radiate into epiphytic niches in angiosperm dominated landscapes. However, our understanding of how ecophysiological function allowed establishment in the canopy and the potential transitionary role of the hemi-epiphytic life form remain unclear. Here, we surveyed 39 fern species in Costa Rican tropical forests to explore epiphytic trait divergence in a phylogenetic context. We examined leaf responses to water deficits in terrestrial, hemi-epiphytic and epiphytic ferns and related these findings to functional traits that regulate leaf water status. Epiphytic ferns had reduced xylem area (-63%), shorter stipe lengths (-56%), thicker laminae (+41%) and reduced stomatal density (-46%) compared to terrestrial ferns. Epiphytic ferns exhibited similar turgor loss points, higher osmotic potential at saturation and lower tissue capacitance after turgor loss than terrestrial ferns. Overall, hemi-epiphytic ferns exhibited traits that share characteristics of both terrestrial and epiphytic species. Our findings clearly demonstrate the prevalence of water conservatism in both epiphytic and hemi-epiphytic ferns, via selection for anatomical and structural traits that avoid leaf water stress. Even with likely evolutionarily constrained physiological function, adaptations for drought avoidance have allowed epiphytic ferns to successfully endure the stresses of the canopy habitat.

Seed predation increases from the Arctic to the Equator and from high to low elevations.

Species interactions have long been predicted to increase in intensity toward the tropics and low elevations because of gradients in climate, productivity, or biodiversity. Despite their importance for understanding global ecological and evolutionary processes, plant-animal interaction gradients are particularly difficult to test systematically across large geographic gradients, and evidence from smaller, disparate studies is inconclusive. By systematically measuring postdispersal seed predation using 6995 standardized seed depots along 18 mountains in the Pacific cordillera, we found that seed predation increases by 17% from the Arctic to the Equator and by 17% from 4000 meters above sea level to sea level. Clines in total predation, likely driven by invertebrates, were consistent across treeline ecotones and within continuous forest and were better explained by climate seasonality than by productivity, biodiversity, or latitude. These results suggest that species interactions play predictably greater ecological and evolutionary roles in tropical, lowland, and other less seasonal ecosystems.

Can fern spores develop hydration memory in response to priming?

Fern spores and seeds initiate germination with fast water uptake, followed by a stationary phase with no appreciable water uptake and biochemical and metabolic processes that precede germination. After that, seed, germination is avoided by dehydration, as part of the priming treatments. After dehydration, seeds maintain their metabolic advances (hydration memory). As a result, rehydrated seeds germinate rapidly. We hypothesized that, as seeds, fern spores may be capable of developing hydration memory. To assess priming, spores of six fern species were exposed to: four or eight days of hydration in water (hydro-priming) or in a soil matrix (matrix-priming); or 1 month of hydration in the soil of the collection site (natural-priming). At the end of the treatments, the spores were dehydrated in the dark and germinated under laboratory conditions. Germination was evaluated using lag-time, germination rate and germination percentage. Priming treatments shortened lag time and/or increased germination rate or germination percentage in relation to the controls. Matrix-priming (8 days) reduced the spore germination percentage in three species. Our results provide evidence that fern spores possess a hydration memory that probably evolved in the soil bank and suggests that hydration-dehydration cycles within the natural soil might provide advantages for successful germination.

A Review of Galls on Ferns and Lycophytes.

Fern-insect interactions have not received the same attention as angiosperm-insect interactions have. It has even been stated that ferns may have very few interactions with animals because of their lack of flowers, fruits, and seeds. Consequently, for many decades fern-insect interactions have been overlooked and underestimated, especially for highly developed interactions such as those with gall-formers. The present work aims to review the galls of ferns and lycophytes worldwide, to provide an updated checklist including unpublished data and to estimate the global gall diversity of ferns and lycophytes. We recorded 93 host species, belonging to 41 genera. Galls were found in 20 fern families and one lycophyte family (Selaginellaceae). Most galls occur within the more derived ferns of the order Polypodiales, especially the fern families Polypodiaceae (21 host species), Dryopteridaceae (14 host species) and Athyriaceae (11 host species). Thirty-eight of the 133 gall morphotypes were induced by mites and 95 by insects of six orders (Coleoptera, Diptera, Hymenoptera, Lepidoptera, Thysanoptera, and Hemiptera). Among the insects, Cecidomyiidae (Diptera) caused most of the galls (35 morphotypes). So far, most galls have been reported from the Neotropical region (40 spp.) and Oriental region (28 spp.).

Geometry, Allometry and Biomechanics of Fern Leaf Petioles: Their Significance for the Evolution of Functional and Ecological Diversity Within the Pteridaceae.

Herbaceous plants rely on a combination of turgor, ground tissues and geometry for mechanical support of leaves and stems. Unlike most angiosperms however, ferns employ a sub-dermal layer of fibers, known as a hypodermal sterome, for support of their leaves. The sterome is nearly ubiquitous in ferns, but nothing is known about its role in leaf biomechanics. The goal of this research was to characterize sterome attributes in ferns that experience a broad range of mechanical stresses, as imposed by their aquatic, xeric, epiphytic, and terrestrial niches. Members of the Pteridaceae meet this criteria well. The anatomical and functional morphometrics along with published values of tissue moduli were used to model petiole flexural rigidity and susceptibility to buckling in 20 species of the Pteridaceae. Strong allometric relationships were observed between sterome thickness and leaf size, with the sterome contributing over 97% to petiole flexural rigidity. Surprisingly, the small-statured cheilanthoid ferns allocated the highest fraction of their petiole to the sterome, while large leaves exploited aspects of geometry (second moment of area) to achieve bending resistance. This pattern also revealed an economy of function in which increasing sterome thickness was associated with decreasing fiber cell reinforcement, and fiber wall fraction. Lastly, strong petioles were associated with durable leaves, as approximated by specific leaf area. This study reveals meaningful patterns in fern leaf biomechanics that align with species leaf size, sterome attributes and life-history strategy.

The evolution of aluminum accumulation in ferns and lycophytes.

PREMISE OF THE STUDY: This paper investigates the occurrence and evolution of aluminum (Al) accumulation within ferns and lycophytes, which is characterized by Al concentrations above 1000 mg·kg-1 in aboveground plant tissues. We hypothesize that this feature is more common in ferns than in angiosperms, and potentially correlated with growth form and other chemical elements. METHODS: Aluminum concentrations were obtained from novel analyses and literature for a total of 354 specimens and 307 species. Moreover, a semiquantitative aluminon test was applied for a subset of 105 species and validated against exact Al measurements. KEY RESULTS: Molecular phylogenetic analyses showed that the major Al-accumulating groups were primarily found in the Gleicheniales and Cyatheales, and largely absent in the Polypodiales. At the species and generic level, Al accumulation was typically either absent or present, and mixed results within a single species and genus were limited to less than 30% of the species and genera tested. Epiphytic ferns had significantly lower Al levels than terrestrial ferns, although this finding was not significant after phylogenetic correction. In addition, a significant, positive correlation was found between Al and iron, while Al was negatively correlated with phosphorus and potassium concentrations. CONCLUSIONS: Aluminum accumulation is most common outside of the Polypodiales and occurs in 38% of the species studied, indicating that this trait is indeed common within subtropical and tropical ferns, a finding that could be in line with their role as pioneer species on landslides and soils with high levels of soluble Al.