Pit membranes in bordered pits of tracheary elements of angiosperm xylem represent primary cell walls that undergo structural and chemical modifications, not only during cell death but also during and after their role as safety valves for water transport between conduits. Cellulose microfibrils, which are typically grouped in aggregates with a diameter between 20 to 30 nm, make up their main component. While it is clear that pectins and hemicellulose are removed from immature pit membranes during hydrolysis, recent observations of amphiphilic lipids and proteins associated with pit membranes raise important questions about drought-induced embolism formation and spread via air-seeding from gas-filled conduits. Indeed, mechanisms behind air-seeding remain poorly understood, which is due in part to little attention paid to the three-dimensional structure of pit membranes in earlier studies. Based on perfusion experiments and modelling, pore constrictions in fibrous pit membranes are estimated to be well below 50 nm, and typically smaller than 20 nm. Together with the low dynamic surface tensions of amphiphilic lipids at air-water interfaces in pit membranes, 5 to 20 nm pore constrictions are in line with the observed xylem water potentials values that generally induce spread of embolism. Moreover, pit membranes appear to show ideal porous medium properties for sap flow to promote hydraulic efficiency and safety due to their very high porosity (pore volume fraction), with highly interconnected, non-tortuous pore pathways, and the occurrence of multiple pore constrictions within a single pore. This three-dimensional view of pit membranes as mesoporous media may explain the relationship between pit membrane thickness and embolism resistance, but is largely incompatible with earlier, two-dimensional views on air-seeding. It is hypothesised that pit membranes enable water transport under negative pressure by producing stable, surfactant coated nanobubbles while preventing the entry of large bubbles that would cause embolism.

Pit membranes between xylem vessels play a major role in angiosperm water transport. Yet, their three‐dimensional (3D) structure as fibrous porous media remains unknown, largely due to technical challenges and sample preparation artefacts. Here, we applied a modelling approach based on thickness measurements of fresh and fully shrunken pit membranes of seven species. Pore constrictions were also investigated visually by perfusing fresh material with colloidal gold particles of known sizes. Based on a shrinkage model, fresh pit membranes showed tiny pore constrictions of ca. 20 nm, but a very high porosity (i.e. pore volume fraction) of on average 0.81. Perfusion experiments showed similar pore constrictions in fresh samples, well below 50 nm based on transmission electron microscopy. Drying caused a 50% shrinkage of pit membranes, resulting in much smaller pore constrictions. These findings suggest that pit membranes represent a mesoporous medium, with the pore space characterized by multiple constrictions. Constrictions are much smaller than previously assumed, but the pore volume is large and highly interconnected. Pores do not form highly tortuous, bent, or zigzagging pathways. These insights provide a novel view on pit membranes, which is essential to develop a mechanistic, 3D understanding of air‐seeding through this porous medium.

Bordered pit membranes of angiosperm xylem are anisotropic, mesoporous media between neighbouring conduits, with a key role in long distance water transport. Yet, their mechanical properties are poorly understood. Here, we aim to quantify the stiffness of intervessel pit membranes over various growing seasons. By applying an AFM-based indentation technique “Quantitative Imaging” we measured the effective elastic modulus (Eeffective) of intervessel pit membranes of Clematis vitalba in dependence of size, age, and hydration state. The indentation-deformation behaviour was analysed with a non-linear membrane model, and paired with magnetic resonance imaging to visualise sap-filled and embolised vessels, while geometrical data of bordered pits were obtained using electron microscopy. Eeffective was transformed to the geometrically independent apparent elastic modulus Eapparent and to aspiration pressure Pb. The material stiffness (Eapparent) of fresh pit membranes was with 57 MPa considerably lower than previously suggested. The estimated pressure for pit membrane aspiration was 2.20+28 MPa. Pit membranes from older growth rings were shrunken, had a higher material stiffness and a lower aspiration pressure than current year ones, suggesting an irreversible, mechanical ageing process. This study provides an experimental-stiffness analysis of hydrated intervessel pit membranes in their native state. The estimated aspiration pressure suggests that membranes are not deflected under normal field conditions. Although absolute values should be interpreted carefully, our data suggest that pit membrane shrinkage implies increasing material stiffness, and highlight the dynamic changes of pit membrane mechanics and their complex, functional behaviour for fluid transport.

Premise: Among the sophisticated trap types in carnivorous plants, the underground eel traps of corkskrew plants (Genlisea spp., Lentibulariaceae) are probably the least understood in terms of their functional principle. Here, we provide a detailed analysis of structural and hydraulic features of G. hispidula traps, contributing to the ongoing debate on whether these traps can actively generate water streams to promote prey capture. Methods: Anatomical and hydraulic traits of detached traps, including inner trap diameters, chamber line element, hair length, glandular pattern, and hydraulic conductivity, were investigated quantitatively using light and electron microscopy, x-ray microtomography, and hydraulic measurements. Results: Hydraulic resistivity in the neck of the trap, from the trap mouth toward the vesicle (digestive chamber) was 10 times lower than in the opposite direction. The comparison of measured and theoretical flow rates suggests that the retrorse hairs inside trap necks also provide considerable resistance against movement of matter toward the vesicle. Hairs showed a gradient in length along the neck, with the shortest hairs near the vesicle. Co-occurrence of quadrifid and bifid glands was limited to a small part of the neck, with quadrifids near the vesicle and bifids toward the trap mouth. Conclusions: The combination of structural gradients with hydraulic anisotropy suggests the trap is a highly fine-tuned system based on likely trade-offs between efficient prey movement in the trap interior toward the vesicle, prey retention, and spatial digestion capacities and is not counter to the generation of water streams.

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