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Conifers dominated wet lowland tropical forests 100 million years ago (MYA). With a few exceptions in the Podocarpaceae and Araucariaceae, conifers are now absent from this biome. This shift to angiosperm dominance also coincided with a large decline in atmospheric CO concentration (c ). We compared growth and physiological performance of two lowland tropical angiosperms and conifers at c levels representing pre-industrial (280 ppm), ambient (400 ppm) and Eocene (800 ppm) conditions to explore how differences in c affect the growth and water-use efficiency (WUE) of seedlings from these groups. Two conifers (Araucaria heterophylla and Podocarpus guatemalensis) and two angiosperm trees (Tabebuia rosea and Chrysophyllum cainito) were grown in climate-controlled glasshouses in Panama. Growth, photosynthetic rates, nutrient uptake, and nutrient use and water-use efficiencies were measured. Podocarpus seedlings showed a stronger (66 %) increase in relative growth rate with increasing c relative to Araucaria (19 %) and the angiosperms (no growth enhancement). The response of Podocarpus is consistent with expectations for species with conservative growth traits and low mesophyll diffusion conductance. While previous work has shown limited stomatal response of conifers to c , we found that the two conifers had significantly greater increases in leaf and whole-plant WUE than the angiosperms, reflecting increased photosynthetic rate and reduced stomatal conductance. Foliar nitrogen isotope ratios (δ N) and soil nitrate concentrations indicated a preference in Podocarpus for ammonium over nitrate, which may impact nitrogen uptake relative to nitrate assimilators under high c SIGNIFICANCE: Podocarps colonized tropical forests after angiosperms achieved dominance and are now restricted to infertile soils. Although limited to a single species, our data suggest that higher c may have been favourable for podocarp colonization of tropical South America 60 MYA, while plasticity in photosynthetic capacity and WUE may help account for their continued persistence under large changes in c since the Eocene.

Background and AimsDespite the importance of vessels in angiosperm roots for plant water transport, there is little research on the microanatomy of woody plant roots. Vessels in roots can be interconnected networks or nearly solitary, with few vessel–vessel connections. Species with few connections are common in arid habitats, presumably to isolate embolisms. In this study, measurements were made of root vessel pit sizes, vessel air-seeding pressures, pit membrane thicknesses and the degree of vessel interconnectedness in deep (approx. 20 m) and shallow (<10 cm) roots of two co-occurring species, Sideroxylon lanuginosum and Quercus fusiformis.MethodsScanning electron microscopy was used to image pit dimensions and to measure the distance between connected vessels. The number of connected vessels in larger samples was determined by using high-resolution computed tomography and three-dimensional (3-D) image analysis. Individual vessel air-seeding pressures were measured using a microcapillary method. The thickness of pit membranes was measured using transmission electron microscopy.Key ResultsVessel pit size varied across both species and rooting depths. Deep Q. fusiformis roots had the largest pits overall (>500 µm) and more large pits than either shallow Q. fusiformis roots or S. lanuginosum roots. Vessel air-seeding pressures were approximately four times greater in Q. fusiformis than in S. lanuginosum and 1·3–1·9 times greater in shallow roots than in deep roots. Sideroxylon lanuginosum had 34–44 % of its vessels interconnected, whereas Q. fusiformis only had 1–6 % of its vessels connected. Vessel air-seeding pressures were unrelated to pit membrane thickness but showed a positive relationship with vessel interconnectedness.ConclusionsThese data support the hypothesis that species with more vessel–vessel integration are often less resistant to embolism than species with isolated vessels. This study also highlights the usefulness of tomography for vessel network analysis and the important role of 3-D xylem organization in plant hydraulic function.

Leaf-Cutting ants (Atta spp.) have become a topical issue in Neotropical ecology, particularly because they are reaching hyper-abundance due to escalating levels of fragmentation in recent years. Yet, despite intensive research on their role as dominant herbivores, there is still insufficient documentation on the impacts of their large, long-lived nests on plant assemblage structure and ecosystem functioning. Our study aimed at investigating the magnitude, nature, and spatial extent of nest influence by assessing 11 attributes of ant nest, canopy structure, light environment and sapling assemblage for 20 colonies in four plots along nest-understorey gradients in a large remnant of Atlantic forest. We also monitored the performance of seeds and seedlings of Chrysophyllum viride, an abundant shade-tolerant species. Previously unrecognized canopy gaps above ant nests (0.04-87.9 m²) occurred in 95% of all colonies surveyed. Overall, canopy openness and light availability at least doubled in ant nest plots compared with distant understorey plots. These drastic changes in the light environment paralleled those in plant assemblage: sapling density almost tripled (mean ± SE: 0.42 ± 0.1 saplings m⁻²) and sapling species richness doubled (0.16 ± 0.02 species m⁻²) in distant plots, as did shade-tolerant species. After a 1-year period, only 33 ± 15.6% of the seeds germinated and all seedlings died on nests, whereas seed germination reached 68 ± 5.1% in distant plots and 66.4 ± 7.6% of their seedlings survived after 12 months. Therefore, plot location was the most significant explanatory variable for predictable and conspicuous changes in the light environment and structure of sapling assemblages. Our findings greatly extend knowledge on the role played by leaf-cutting ants as ecosystem engineers by demonstrating that ant nest-mediated disturbance promotes environmental modifications in tens of meters around nests and is thus, strong enough to drive plant recruitment and consequently alter both the floristic and functional signature of plant assemblages.

Chrysophyllum albidum Linn (African star apple) is a fruit with extensive nutritional and medicinal benefits. The fruit and kernel in the seed are both edible. Strains of lactic acid bacteria (LAB) were isolated from fermented seeds and assessed for probiotic characteristics. The extracts in both the unfermented and the fermented aqueous extracts from the kernels obtained from the seeds of C. albidum were subjected to analysis using the gas chromatography/mass spectrometry (GC-MS) method. This analysis identified the bioactive compounds present as possible substrate(s) for the associated organisms inducing the fermentation and the resultant biotransformed products formed. Three potential probiotic LAB strains identified as Lactococcus raffinolactis (ProbtA1), Lactococcus lactis (ProbtA2a), and Pediococcus pentosaceus (ProbtA2b) were isolated from the fermented C. albidum seeds. All strains were non hemolytic, which indicated their safety, Probt (A1, A2a, and A2b) grew in an acidic environment (pH 3.5) during the 48-h incubation time, and all three strains grew in 1% bile, and exhibited good hydrophobicity and auto-aggregation properties. Mucin binding proteins was not detected in any strain, and bile salt hydrolase was detected in all the strains. l-lactic acid (28.57%), norharman (5.07%), formyl 7E-hexadecenoate (1.73%), and indole (1.51%) were the four major constituents of the fermented kernel of the C. albidum, while 2,5-dimethylpyrazine (C1, 1.27%), 3,5-dihydroxy-6-methyl-2,3-dihydropyran-4-one (C2, 2.90%), indole (C3, 1.31%), norharman (C4, 3.01%), and methyl petroselinate (C5, 4.33%) were the five major constituents of the unfermented kernels. The isolated LAB are safe for consumption. The fermenting process metabolized C1, C2, and C5, which are possible starter cultures for the growth of probiotics. Fermentation is an essential tool for bioengineering molecules in foods into safe and health beneficial products.

Ethnobotanical studies have reported the traditional medicinal uses of Acacia senegal (L.) Willd. and Argania spinosa (L.) Skeels against kidney stone formation and other chronic kidney diseases. The present work is undertaken to study the litholytic activity and the inhibiting activity of calcium oxalate crystallization by bioactive compounds identified in Argania spinosa (L.) Skeels press-cake (residue of Argan oil) and in Acacia senegal (L.) Willd. The litholytic activity was studied in vitro on cystine and uric acid stones using a porous bag and an Erlenmeyer glass. The study of the inhibiting activity of calcium oxalate crystallization, was based on temporal measurements of the optical density, registered at a 620 nm wavelength for 30 min using an ultraviolet–visible spectrophotometer. The silylation method was performed to identify phytochemicals, followed by gas chromatography coupled with mass spectrophotometry (GC/MS) analysis. The results show significant litholytic activity of Argania Spinosa press-cake hydro-ethanolic extract on uric acid and cystine stones, respectively, with dissolution rates (DR) of 86.38% and 60.42% versus 3.23% and 9.48% for the hydro-ethanolic extract of Acacia senegal exudate. Furthermore, the percentages of nucleation inhibition are 83.78% and 43.77% (p ˂ 0.05) for Argania spinosa and Acacia senegal, respectively. The results point to the detection of 17 phytochemicals in Argania spinosa press-cake extract, the majority of which are phenolic acids and have potent anti-urolithiatic action.

Previous studies have determined that Chloroluma gonocarpa (Sapotaceae), is a species that has cryptic dioecy. This type of sexual system is characterized by flowers that are morphologically perfect (both sexual whorls are present) but functionally pistillate or staminate (in each type of flower one of the sexual whorls is non-functional). In C. gonocarpa the pistillate flowers present well-developed stigma, functional ovules, and staminodes, while the staminate flowers present a poorly developed stigma, collapsed ovules, and pollen-producing anthers. In angiosperms, the abortion of sexual organs can occur at different stages of development (from pre-meiosis to post-meiosis), that is why we conducted an anatomical analysis of both flower types at various developmental stages. Using light microscopy, we described the processes of sporogenesis and gametogenesis to establish when the staminate flowers lose their pistillate function. To achieve this, we collected, fixed, and processed the flowers following conventional anatomical techniques for observation under a light microscope. Our findings reveal that pollen development occurs only in staminate flowers, while ovule development begins in both types of flowers but ceases in staminate flowers due to post-meiosis abortion. In contrast, normal development continues in pistillate flowers. These results suggest that dioecy in C. gonocarpa may have arisen from a gynodioecious pathway.

The mycorrhizal fungi of cultivated Vanilla spp. have mainly been studied in America, while a recent study has investigated them on Réunion Island (Indian Ocean). However, there are many different types of cultivation on Réunion, from shade-house crops to forest farms of endemic or exotic trees. Here we fill a gap in the study of the root mycobiome of Vanilla by sampling vines in forest plantations on recent lava flows in the southeast of Réunion. Specifically, we aimed to characterize the fungal communities between terrestrial and epiphytic roots, between forest farms that differ mainly in the species of trees, and between Vanilla roots and ECM-like roots of nearby trees. By sequencing fungal ITS2, we showed that the Vanilla root mycobiome is diverse and differed between the root types and forest farms. Epiphytic and terrestrial roots host endophytic fungi, while a putative rust with visible urediniospores was abundant in terrestrial roots mainly. Other pathogens were detected in epiphytic roots (Colletotrichum) with no sign of disease. Following sequencing and electron microscopy, Tulasnellaceae, characterized by imperforate parenthesomes and cell wall expansion with an amorphous matrix, were shown to be the main mycorrhizal fungi in both vanilla root types. Interestingly, the dominant Tulasnellaceae OTU was found in ECM-type roots of trees belonging to the ectomycorrhizal family Sapotaceae. Further observations are needed to confirm the ectomycorrhizal association of endemic trees with Tulasnella. Moreover, labeling experiments will be instrumental in investigating the transfer of nutrients between the trees and the Vanilla through the network of mycorrhizal associations in the soil.

Indigenous fruit tree species (IFTs) can play a crucial role in poverty alleviation and as a source of food in South Africa. Although these species contain nutrient-rich edible fruits that are also locally processed to food products such as jams and jellies, they are still underutilized and also harvested from the wild. This study aimed to prioritize in northern KwaZulu-Natal, IFTs with domestication and commercialization potential. A literature survey and focus groups were used to list tree species that produce edible fruits with this potential. Plant experts, through focus groups and reference group meetings, further ranked fruit trees using a scorecard method based on various attributes. A total of 29 prioritized IFTs was obtained, of which the Sapotaceae was the most represented family with a maximum of four species. Based on scorecard assessment, the most preferred species in descending order were Strychnos spinosa Lam., Garcinia livingstonei T.Anderson, Englerophytum magalismontanum (Sond.) T.D.Penn, Sclerocarya birrea (A.Rich) Hochst., Dovyalis caffra (Hook.f. & Harv) Hook.f., Vangueria infaustia Burch. and Berchemia zeyheri (Sond.) Grubov. Strychnos spinosa was ranked as the first most important IFTs with a final score of 151, whereas Berchemia zeyheri was ranked the least with the final score of 146.1. However, the final scores of all most preferred species were relatively similar to one another. The top seven species identified in this study should be considered as a baseline for future IFTs development programmes such as domestication, whereby these species are introduced to small-holder farmers and food processing industries.

Aims This study aimed to establish elemental profiles and to spatially resolve the elemental distribution in five New Caledonian woody Ni hyperaccumulator plant species ( Geissois pruinosa var . pruinosa , Homalium francii , Hybanthus austrocaledonicus , Psychotria gabriellae , and Pycnandra acuminata ) originating from the Cunoniaceae, Salicaceae, Violaceae, Rubiaceae, and Sapotaceae families respectively. Methods Using synchrotron-based micro-X-ray Fluorescence (μXRF) imaging of different plant tissues, from the roots to the shoots and reproductive organs, this study aimed to clarify how distribution patterns of nickel, and other physiologically relevant elements, differ between these species. Results The results show that the tissue-level and cellular-level distribution of nickel in P. gabriellae, H. austrocaledonicus, G. pruinosa var . pruinosa, and H. francii conform with the majority of studied Ni hyperaccumulator plant species globally, including (temperate) herbaceous species, with localization mainly in epidermal cells and phloem bundles. However, P. acuminata has nickel-rich laticifers, which constitute an independent network of cells that is parallel to the vascular bundles and are the main sink for nickel. Conclusions Synchrotron-based micro-X-ray Fluorescence (μXRF) is a powerful method for investigating how metal hyperaccumulation influences acquisition and spatial distribution of a wide range of elements. This non-invasive method enables investigation into the in vivo distribution of multiple elements and the structure and organisation of cells (e.g. laticifers).

The tree-shrub species Pouteria glomerata (Miq.) Radlk. (Sapotaceae) occurs in periodically flooded areas marked by periods of drought. It has the potential for use in the ecological restoration of riparian forests; however, seed germination strategies were not fully understood. In this study, we aimed to evaluate the morphology, anatomy, and physiological aspects of the seed to identify possible factors related to low germination. The morphoanatomical seed analysis was assessed according to the routine procedures. For physiological assessment, seed germination was estimated under different conditions; seeds were scarified and soaked in water to overcome dormancy. Seeds store starch, lipids, and proteins, and are covered by a hard and thick seed coat. The embryo was underdeveloped consisting of a mass of meristematic cells. Numerous laticifers and phenolic idioblasts were found, mostly on the periphery of the cotyledons. Germination was hypogeal and the seedling was cryptocotylar type. The treatments to overcome seed dormancy were not efficient to break dormancy. The morphoanatomical analysis suggested that the dormancy of P. glomerata seeds may be related to an impermeable seed coat and undifferentiated or immature embryo indicating a morphophysiological dormancy.