Biology subjects

Holbrook, N. M.

Publications and source records attributed to Holbrook, N. M..

13 recordsLinked to original sources

bioRxiv2026

Pectin is a critical contributor to the mechanical properties of Chara cell walls

Cellulose has long been viewed as the major load-bearing component in plant primary cell walls. Recent studies, however, suggest that pectin plays a significant role in cell wall mechanics despite a kPa-scaled modulus. Here we quantify the mechanical properties of centimeter-sized internodal cell walls of Chara corallina following treatments to remove calcium crosslinks or digest pectin. We also characterize the flow of water and gas through the cell wall as a function of pressure. We found that removal of calcium crosslinks did not affect wall strength and stress-relaxation behavior, yet it decreased the elastic modulus and increased permeability to water. In contrast, pectin removal had a dramatic impact on all measurements. Samples from which pectin had been removed exhibited lower strength and stiffness and had greater stress relaxation under constant strain. Pectin removal increased the permeability of the cell wall to water above that observed when calcium was removed and reduced the pressure at which gas "tunnels" through the cell wall ~ 2-fold. Our findings support the idea that pectin locally restricts the movement of cellulose microfibrils, with implications for understanding functional properties of primary cell walls in vascular plants and developing high-performance double-network materials.

biophysics↗

bioRxiv2026

The origin of mechanical advantage in angiosperms

O_LIMechanical interaction between guard cells and epidermal pavement cells enables large stomatal apertures and high productivity in angiosperms. We do not know when this response evolved, but over the last 169 years we have found that mechanical advantage has been tested in at least 230 species from 85 families. To date no data on this trait exists among angiosperms outside magnoliids, monocots and eudicots. C_LIO_LITo resolve the evolutionary origins of this critical stomatal response we tested for mechanical advantage across 14 additional species including the earliest diverging lineages of angiosperms. C_LIO_LIWe find that mechanical advantage, while variable in magnitude, is present in all angiosperm species that have been measured, including Amborella trichopoda sister to all angiosperms. C_LIO_LIThis response likely evolved once in flowering plants, in the common ancestor of this clade, remaining widespread across angiosperms today. We hypothesize that angiosperms could not have realized the full potential of physiological innovations in water transport without the evolution of this key trait that increased operational stomatal aperture. C_LI

plant biology↗

bioRxiv2025

Secreting salt glands constrain cuticle fracture to enhance desalination efficiency

Plants responding to excessive soil salinity by discharging brine onto their leaf surface risk dehydration through the osmotic continuity between the living tissue and the surface brine, which further enriches with evaporation. Cuticle cracks have long been identified as essential for salt to reach the leaf surface but provide the potentially desiccating continuity between the brine and the gland interior. Using the secreting salt gland of Nolana mollis as a model system, we integrate mathematical modeling, imaging, and physiological measurements to examine the mechanical and biochemical processes required for efficient desalination. We find that the subcuticular space between the concentrated surface brine and the more dilute secreting cell eases the energetic limits of active desalination by reducing the concentration gradient of salt across the cell membrane. We show that crack size plays a critical role in balancing the osmotic and pressure gradients required for salt removal without runaway foliar desiccation.

plant biology↗

bioRxiv2025

Loss of plasma membrane conductance in outside-xylem zone explains non-stomatal control of transpiration

The conventional assumption is that stomatal conductance (gs) dominates the regulation of water and carbon dioxide fluxes between leaves and the atmosphere. Here, a nanoreporter of water status at the mesophyll cell surface and local xylem within intact maize leaves documents significant undersaturation of water vapor in the outside-xylem zone (OXZ) and a large loss of conductance of this zone (goxz) at moderate xylem water stress (no turgor loss). The ratio of the resistances (1/goxz)/(1/gs) serves as a predictive phenotype of undersaturation, non-stomatal regulation of transpiration, errors in standard gas exchange analysis, and an increase of intrinsic water use efficiency (iWUE). Cell-scale access to water status reveals symplasmic-apoplasmic disequilibrium and informs a biophysical model that can explain experimental observations quantitatively based on localization of variable conductance to the plasma membrane. This work opens new paths of inquiry into the molecular basis and functional consequences of non-stomatal regulation of transpiration.

plant biology↗

bioRxiv2024

Relieving the transfusion tissue traffic jam: a network model of radial transport in conifer needles

The linear geometry of conifer leaves (e.g., pine needles) imposes architectural constraints on solute transport. The needles structural solution to prevent axial stagnation, however, introduces an additional challenge to radial transport by restricting loading and unloading of sugar and water, respectively, to a narrow zone at the periphery of the vascular bundle. Moreover, a Casparian strip blocks apoplastic flow through the endodermis between the vasculature and photosynthetic tissue, forcing countercurrents of water and sugar to travel simultaneously through the cell lumen at this interface. In between these two potential bottlenecks is the transfusion tissue, a distinctive anatomical feature of conifer needles. Here we develop a network-based mathematical model to explore how the structure of the intervening transfusion tissue facilitates radial transport of sugar and water. To describe extravascular transport with cellular resolution, we construct networks from images of Pinus pinea needles obtained through X-ray CT, as well as fluorescence and electron microscopy. Our results show that the physical separation of sugar and water pathways within the transfusion tissue mitigates the consequences of constricting flow at both the vascular access points and the endodermis. SIGNIFICANCEThe efficiency with which plants transport water and sugar across many scales affects their survival and success. At the leaf scale, accommodating the opposing flows of water and sugar is a nontrivial challenge, especially when foliar geometry imposes architectural limitations (e.g., in linear leaves (needles) of conifers). The transfusion tissue is a characteristic component of the conifer needle which mediates the radial transport of sugar and water. Our network model shows that the transfusion tissue alleviates the sugar-water traffic jams that would otherwise develop between the vascular system and photosynthetic cells. Our mathematical model provides finer resolution than what is currently possible with experimental approaches.

plant biology↗

bioRxiv2023

Wind speed affects the rate and kinetics of stomatal conductance

Understanding the relationship between wind speed and gas exchange in plants is a longstanding challenge. Our aim was to investigate the impact of wind speed on maximum rates of gas exchange and the kinetics of stomatal responses. We conducted experiments using an infrared gas analyzer equipped with a controlled leaf fan, enabling precise control of the boundary layer conductance. We first showed that the chamber was adequately mixed even at extremely low fan speeds (down to 200 rpm, equivalent to a wind speed of 0.0005 m s-1) and evaluated the link between fan speed, wind speed, and boundary layer conductance. We observed that higher wind speeds led to increased gas exchange of both water vapor and CO2 in Arabidopsis, presumably due to its effect on transpiration and the consequential reduction in epidermal pressure that led to stomatal opening. We documented that stomatal opening in response to light was three times faster at a fan speed of 10000 rpm (wind speed of 2 m s-1) compared with 500 rpm (0.25 m s-1) in Vicia faba, while the latter exhibited an opening rate that was similar to those of epidermal peels. The increase of stomatal conductance under high wind was observed in four species under field conditions. Our findings demonstrate the importance of the size of the boundary layer on determining maximum rates of gas exchange and the kinetics of gas exchange responses to environmental changes.

plant biology↗

bioRxiv2023

Localized measurements of water potential reveal large loss of conductance in living tissues of maize leaves

The water status of the living tissue in leaves between the xylem and stomata (outside xylem zone - OXZ) play a critical role for plant function and global mass and energy balance but has remained largely inaccessible. We resolve the local water relations of OXZ tissue using a nanogel reporter of water potential ({psi}), AquaDust, that enables an in-situ, non-destructive measurement of both{psi} of xylem and highly localized{psi} at the terminus of transpiration in the OXZ. Working in maize, these localized measurements reveal gradients in the OXZ that are several fold larger than those based on conventional methods, and values of{psi} in the mesophyll apoplast well below the macroscopic turgor loss potential. We find a strong loss of hydraulic conductance in both the bundle sheath and the mesophyll with decreasing xylem potential but not with evaporative demand. Our measurements suggest an active role played by the OXZ in regulating the transpiration path and our methods provide novel means to study this phenomenon.

physiology↗

bioRxiv2021

Sieve tube structural variation in Austrobaileya scandens and its significance for lianescence

Lianas are characterized by large leaf areas and slender stems, a combination of features that require an efficient vascular system. The only extant member of the Austrobaileyaceae, is an endemic twining liana of the tropical Australian forests with well-known xylem hydraulic traits. However, the vascular phloem continuum through aerial organs remains understudied. We analyzed the structure of phloem conduits across leaf veins and stems of A. scandens, combining topological data obtained through light and electron microscopy, with current models of phloem transport. Leaves displayed a low xylem to phloem ratio compared with leaves of other angiosperms, with vascular elements invariant in diameter along the midrib, but tapered across vein hierarchies. Sieve plate pore radii were extremely small: 0.08 {micro}m in minor veins, increasing to 0.12{micro}m in the petiole and only to 0.20{micro}m at the base of the stem, tens of meters away. Searcher branches contained tube shaped phloem conduits with a pectin-rich wall, whereas twining stems displayed sieve elements with tangential connections that displayed a greater fraction of the tubes populated with an astonishing number of sieve plates. Hydraulic segmentation of the leaves in Austrobaileyaceae correlate with vesseless leaves that benefit photoassimilate export through volumetric scaling of the sieve tube elements. Yet, compared with canopy dominant trees, the geometrical properties of the sieve tube in twining stems, restrict considerably energy distribution in the sub-canopy layers, potentially favoring the allocation of assimilates toward the elongating branches. Thus, the conductive xylem of twining stems contrasts with a poorly conductive phloem that meets the mechanical constraints of lianescence.

plant biology↗

bioRxiv2021

Changes In Ploidy Affect Vascular Allometry And Hydraulic Function In Trees

O_LIThe enucleated vascular elements of the xylem and the phloem offer an excellent system to test the effect of ploidy on plant function because variation in vascular geometry has a direct influence on transport efficiency. However, evaluations of conduit sizes in polyploid plants have remained elusive, most remarkably in woody species. C_LIO_LIWe used a combination of molecular, physiological, and microscopy techniques to model the hydraulic resistance between source and sinks in tetraploid and diploid mango trees. C_LIO_LITetraploids exhibited larger chloroplasts, mesophyll cells, and stomatal guard cells, resulting in higher leaf elastic modulus and lower dehydration rates despite the high water potentials of both ploidies in the field. Both the xylem and the phloem displayed a scaling of conduits with ploidy, revealing attenuated hydraulic resistance in tetraploids. C_LIO_LIConspicuous wall hygroscopic moieties in the cells involved in processes of transpiration and transport advocates a role in volumetric adjustments due to turgor change in polyploids, which, together with the enlargement of organelles, cells, and tissues that are critical for water and photo assimilate transport at long distances, imply major physiological novelties of polyploidy. C_LI

plant biology↗

bioRxiv2021

Intergenerational niche differentiation in filmy ferns (Hymenophyllaceae)

Desiccation tolerance was a key trait that allowed plants to colonize land. However, little is known about the transition from desiccation tolerant non-vascular plants to desiccation sensitive vascular ones. Filmy ferns (Hymenophyllaceae) represent a useful system to investigate how water-stress strategies differ between non-vascular and vascular stages within a single organism because they have vascularized sporophytes and nonvascular gametophytes that are each capable of varying degrees of desiccation tolerance. To explore this, we surveyed sporophytes and gametophytes of 19 species (22 taxa including varieties) of filmy ferns on Moorea (French Polynesia) and used chlorophyll fluorescence to measure desiccation tolerance and light responses. We conducted phylogenetically informed analyses to identify differences in physiology between life stages and growth habits. Gametophytes had similar or less desiccation tolerance (ability to recover from 2 d desiccation at -86 MPa) and lower photosynthetic optima (maximum electron transport rate of photosystem II and light level at 95% of that rate) than sporophytes. Epiphytes were more tolerant of desiccation than terrestrial species in both life stages. Despite their lack of greater physiological tolerances, gametophytes of several species occurred over a wider elevational range than conspecific sporophytes. Our results demonstrate that filmy fern gametophytes and sporophytes differ in their physiology and niche requirements, and point to the importance of microhabitat in shaping the evolution of water-use strategies in vascular plants.

ecology↗

bioRxiv2019

Idioblasts as pathways for distributing water absorbed by leaf surfaces to the mesophyll in Capparis odoratissima

Capparis odoratissima is a tree species native to semi-arid environments of the northern coast of South America where low soil water availability coexists with frequent nighttime fog. Previous work with this species demonstrated that C. odoratissima is able to use water absorbed through its leaves at night to enhance leaf hydration, photosynthesis, and growth. Here, we combine detailed anatomical evaluations of the leaves of C. odoratissima, with water and dye uptake experiments in the laboratory. We used immunolocalization of pectin and arabinogalactan protein epitopes to characterize the chemistry of foliar water uptake pathways. The abaxial surfaces of C. odoratissima leaves are covered with overlapping, multicellular peltate hairs, while the adaxial surfaces are glabrous but with star-shaped "structures" at regular intervals. Despite these differences in anatomy, both surfaces are able to absorb condensed water, but this ability is most significant on the upper surface. Rates of evaporative water loss from the upper surface, however, are coincident with cuticle conductance. Numerous idioblasts connect the adaxial leaf surface and the adaxial peltate hairs, which contain hygroscopic substances such as arabinogalactan proteins and pectins. The highly specialized anatomy of the leaves of C odoratissima fulfills the dual function of avoiding excessive water loss due to evaporation, while maintaining the ability to absorb liquid water. Cell-wall related hygroscopic compounds present in the peltate hairs and idioblasts create a network of microchannels that maintain leaf hydration and promote the uptake of aerial water.

plant biology↗

bioRxiv2019

Wood capacitance is related to water content, wood density, and anatomy across 30 temperate tree species

Water released from wood tissue during transpiration (capacitance) can meaningfully affect daily water use and drought response. To provide context for better understanding of capacitance mechanisms, we investigated links between capacitance and wood anatomy. On twig wood of 30 temperate angiosperm tree species, we measured capacitance, water content, wood density, and anatomical traits, i.e., vessel dimensions, tissue fractions, and vessel-tissue contact fractions (fraction of vessel circumference in contact with other tissues). Across all species, the strongest predictors of capacitance were wood density (WD) and predawn lumen volumetric water content (VWCL-pd, r2adj=0.44, P<0.0001). Vessel-tissue contact fractions explained an additional [~]10% of the variation in capacitance. Regression models were not improved by including predawn relative water content (RWCpd) or tissue lumen fractions. Among diffuse-porous species, VWCL-pd and vessel-ray contact fraction were the best predictors of capacitance, whereas among ring/semi-ring-porous species, VWCL-pd, WD and vessel-fibre contact fraction were the best predictors. Mean RWCpd was 0.65{+/-}0.13 and uncorrelated with WD. VWCL-pd was weakly negatively correlated with WD. Our findings imply that capacitance depends on the amount of stored water, tissue connectivity and the bulk wood properties arising from WD (e.g., elasticity), rather than the fraction of any particular tissue.

plant biology↗

bioRxiv2018

Phloem structure and development in Illicium parviflorum, a basal angiosperm shrub

Recent studies in canopy-dominant trees revealed a structure-function scaling of the phloem. However, whether axial scaling is conserved in woody plants of the understory, the environments of most basal-grade angiosperms, remains mysterious. We used seedlings and adult plants of the shrub Illicium parviflorum to explore the anatomy and physiology of the phloem in their aerial parts, and possible changes through ontogeny. Adult plants maintain a similar proportion of phloem tissue across stem diameters, but scaling of conduit dimensions and number decreases the hydraulic resistance towards the base of the plant. Yet, the small sieve plate pores resulted in an overall higher sieve tube hydraulic resistance than has been reported in other woody angiosperms. Sieve elements scaled from minor to major leaf veins, but were shorter and narrower in petioles. The low carbon assimilation rates of seedlings and mature plants contrasted with a three-fold higher phloem sap velocity in seedlings, suggesting that phloem transport velocity is modulated through ontogeny. While the overall architecture of the phloem tissue in basal-angiosperm understory shrubs scales in a manner consistent with trees, modification of conduit connections may have allowed woody angiosperms to extend beyond their understory origins.

physiology↗