N. M. Holbrook

Optimal vein density in artificial and real leaves

The long evolution of vascular plants has resulted in a tremendous variety of natural networks responsible for the evaporatively driven transport of water. Nevertheless, little is known about the physical principles that constrain vascular architecture. Inspired by plant leaves, we used microfluidic devices consisting of simple parallel channel networks in a polymeric material layer, permeable to water, to study the mechanisms of and the limits to evaporation-driven flow. We show that the flow rate through our biomimetic leaves increases linearly with channel density (1/d) until the distance between channels (d) is comparable with the thickness of the polymer layer (δ), above which the flow rate saturates. A comparison with the plant vascular networks shows that the same optimization criterion can be used to describe the placement of veins in leaves. These scaling relations for evaporatively driven flow through simple networks reveal basic design principles for the engineering of evaporation–permeation-driven devices, and highlight the role of physical constraints on the biological design of leaves.

Leaf palmate venation and vascular redundancy confer tolerance of hydraulic disruption.

Leaf venation is a showcase of plant diversity, ranging from the grid-like network in grasses, to a wide variety of dendritic systems in other angiosperms. A principal function of the venation is to deliver water; however, a hydraulic significance has never been demonstrated for contrasting major venation architectures, including the most basic dichotomy, “pinnate” and “palmate” systems. We hypothesized that vascular redundancy confers tolerance of vein breakage such as would occur during mechanical or insect damage. We subjected leaves of woody angiosperms of contrasting venation architecture to severing treatments in vivo, and, after wounds healed, made detailed measurements of physiological performance relative to control leaves. When the midrib was severed near the leaf base, the pinnately veined leaves declined strongly in leaf hydraulic conductance, stomatal conductance, and photosynthetic rate, whereas palmately veined leaves were minimally affected. Across all of the species examined, a higher density of primary veins predicted tolerance of midrib damage. This benefit for palmate venation is consistent with its repeated evolution and its biogeographic and habitat distribution. All leaves tested showed complete tolerance of damage to second- and higher-order veins, demonstrating that the parallel flow paths provided by the redundant, reticulate minor vein network protect the leaf from the impact of hydraulic disruption. These findings point to a hydraulic explanation for the diversification of low-order vein architecture and the commonness of reticulate, hierarchical leaf venation. These structures suggest roles for both economic constraints and risk tolerance in shaping leaf morphology during 130 million years of flowering plant evolution.

Optimality of the Münch Mechanism for Translocation of Sugars in Plants

Plants require effective vascular systems for the transport of water and dissolved molecules between distal regions. Their survival depends on the ability to transport sugars from the leaves where they are produced to sites of active growth; a flow driven, according to the Münch hypothesis, by osmotic gradients generated by differences in sugar concentration. The length scales over which sugars are produced (Lleaf) and over which they are transported (Lstem), as well as the radius r of the cylindrical phloem cells through which the transport takes place, vary among species over several orders of magnitude; a major unsettled question is whether the Münch transport mechanism is effective over this wide range of sizes. Optimization of translocation speed predicts a scaling relation between radius r and the characteristic lengths as r ∼ (Lleaf Lstem)1/3. Direct measurements using novel in vivo techniques and biomimicking microfluidic devices support this scaling relation and provide the first quantitative support for a unified mechanism of sugar translocation in plants spanning several orders of magnitude in size. The existence of a general scaling law for phloem dimensions provides a new framework for investigating the physical principles governing the morphological diversity of plants.

Ion induced changes in the structure of bordered pit membranes

Ion-mediated changes in xylem hydraulic resistance are hypothesized to result from hydrogel like properties of pectins located in the bordered pit membranes separating adjacent xylem vessels. Although the kinetics of the ion-mediated changes in hydraulic resistance are consistent with the swelling/deswelling behavior of pectins, there is no direct evidence of this activity. In this report we use atomic force microscopy (AFM) to investigate structural changes in bordered pit membranes associated with changes in the ionic concentration of the surrounding solution. When submerged in de-ionized water, AFM revealed bordered pit membranes as relatively smooth, soft, and lacking any sharp edges surface, in contrast to pictures from scanning electron microscope (SEM) or AFM performed on air-dry material. Exposure of the bordered pit membranes to 50 mM KCl solution resulted in significant changes in both surface physical properties and elevation features. Specifically, bordered pit membranes became harder and the fiber edges were clearly visible. In addition, the membrane contracted and appeared much rougher due to exposed microfibers. In neither solution was there any evidence of discrete pores through the membrane whose dimensions were altered in response to the ionic composition of the surrounding solution. Instead the variable hydraulic resistance appears to involve changes in the both the permeability and the thickness of the pit membrane.

Modeling the hydrodynamics of phloem sieve plates

Sieve plates have an enormous impact on the efficiency of the phloem vascular system of plants, responsible for the distribution of photosynthetic products. These thin plates, which separate neighboring phloem cells, are perforated by a large number of tiny sieve pores and are believed to play a crucial role in protecting the phloem sap from intruding animals by blocking flow when the phloem cell is damaged. The resistance to the flow of viscous sap in the phloem vascular system is strongly affected by the presence of the sieve plates, but the hydrodynamics of the flow through them remains poorly understood. We propose a theoretical model for quantifying the effect of sieve plates on the phloem in the plant, thus unifying and improving previous work in the field. Numerical simulations of the flow in real and idealized phloem channels verify our model, and anatomical data from 19 plant species are investigated. We find that the sieve plate resistance is correlated to the cell lumen resistance, and that the sieve plate and the lumen contribute almost equally to the total hydraulic resistance of the phloem translocation pathway.

Environmental and physiological influences on carbon isotope composition of gap and understory plants in a lowland tropical forest.

The stable carbon isotope composition of plant tissue is determined by the isotopic composition of the CO 2 source and discrimination against the heavier isotope 13 C during photosynthetic CO 2 fixation. In C 3 plants, this discrimination has two main components: one associated with diffusion of CO 2 through the stomata and one because of the discrimination against 13 C by the primary carboxylating enzyme RuBisCO. The balance between carboxylation and stomatal limitation of CO 2 diffusion is reflected in the ratio of intercellular to atmospheric partial pressure of CO 2 ( p i / p a ). In a study discussed in this chapter, the foliar δ 13 C and photosynthetic gas exchange of high light-requiring species growing in gaps and of shade-tolerant shrub species growing in the understory and in gaps in a lowland tropical forest were examined. The study compared the leaf gas exchange responses of shade-tolerant shrub species growing in the understory and the same species growing in gaps and comparing gas exchange responses of shade-tolerant and high light-requiring species both growing in gaps. The chapter determines the extent to which the variation in foliar δ 13 C observed could be attributed to differences in internal physiological features or to differences in the isotopic composition of the source CO 2 . The δ 13 C of the air was estimated from the ambient CO 2 concentration.

Leaf physiology does not predict leaf habit; examples from tropical dry forest

Leaf structure and physiology are thought to be closely linked to leaf longevity and leaf habit. Here we compare the seasonal variation in leaf hydraulic conductance (kleaf) and water potential of two evergreen tree species with contrasting leaf life spans, and two species with similar leaf longevity but contrasting leaf habit, one being deciduous and the other evergreen. One of the evergreen species, Simarouba glauca, produced relatively short-lived leaves that maintained high hydraulic conductance year round by periodic flushing. The other evergreen species, Quercus oleoides, produced longer-lived leaves with lower kleaf and as a result minimum leaf water potential was much lower than in S. glauca (−2.8 MPa vs −1.6 MPa). Associated with exposure to lower water potentials, Q. oleoides leaves were harder, had a higher modulus of elasticity, and were less vulnerable to cavitation than S. glauca leaves. Both species operate at water potentials capable of inducing 20 (S. glauca) to 50% (Q. oleoides) loss of kleaf during the dry season although no evidence of cumulative losses in kleaf were observed in either species suggesting regular repair of embolisms. Leaf longevity in the deciduous species Rhedera trinervis is similar to that of S. glauca, although maximum kleaf was lower. Furthermore, a decline in leaf water potential at the onset of the dry season led to cumulative losses in kleaf in R. trinervis that culminated in leaf shedding.

Freeze/Thaw-Induced Embolism: Probability of Critical Bubble Formation Depends on Speed of Ice Formation

Bubble formation in the conduits of woody plants sets a challenge for uninterrupted water transportation from the soil up to the canopy. Freezing and thawing of stems has been shown to increase the number of air-filled (embolized) conduits, especially in trees with large conduit diameters. Despite numerous experimental studies, the mechanisms leading to bubble formation during freezing have not been addressed theoretically. We used classical nucleation theory and fluid mechanics to show which mechanisms are most likely to be responsible for bubble formation during freezing and what parameters determine the likelihood of the process. Our results confirm the common assumption that bubble formation during freezing is most likely due to gas segregation by ice. If xylem conduit walls are not permeable to the salts expelled by ice during the freezing process, osmotic pressures high enough for air seeding could be created. The build-up rate of segregated solutes in front of the ice-water interface depends equally on conduit diameter and freezing velocity. Therefore, bubble formation probability depends on these variables. The dependence of bubble formation probability on freezing velocity means that the experimental results obtained for cavitation threshold conduit diameters during freeze/thaw cycles depend on the experimental setup; namely sample size and cooling rate. The velocity dependence also suggests that to avoid bubble formation during freezing trees should have narrow conduits where freezing is likely to be fast (e.g., branches or outermost layer of the xylem). Avoidance of bubble formation during freezing could thus be one piece of the explanation why xylem conduit size of temperate and boreal zone trees varies quite systematically.

Seedling growth in conifers and angiosperms: impacts of contrasting xylem structure

Competitive interaction between conifers and angiosperms has moulded the structure of global vegetation since the Cretaceous. Angiosperms appear to enjoy their greatest advantage in the lowland tropics, an advantage often attributed to the presence of vessels in their xylem tissue. By monitoring the seedling growth of three members of the pan-tropical conifer family Podocarpaceae and three tropical angiosperm tree species, our aim was to determine whether these conifer and angiosperm seedlings showed distinct patterns of growth and light adaptation that might be attributed to the presence/absence of vessels. Angiosperm seedlings were consistently more efficient in terms of leaf area carried per unit stem investment, as well as more responsive to light climate than the conifer seedlings. Apparently linked to this were larger growth rate, stem hydraulic conductivity and stomatal conductance in the angiosperm sample. Stem hydraulic conductivity and maximum stomatal conductance were highly correlated among species and light treatments explaining the association between highly conductive vessel-bearing wood and high rates of gas exchange. We conclude that xylem vessels contribute to higher rates of gas exchange and more efficient production of leaf area in our sample angiosperms than in conifers. However, this advantage is limited by shade.

Structural and hydraulic correlates of heterophylly in Ginkgo biloba

This study investigates the functional significance of heterophylly in Ginkgo biloba, where leaves borne on short shoots are ontogenetically distinct from those on long shoots. Short shoots are compact, with minimal internodal elongation; their leaves are supplied with water through mature branches. Long shoots extend the canopy and have significant internodal elongation; their expanding leaves receive water from a shoot that is itself maturing. Morphology, stomatal traits, hydraulic architecture, Huber values, water transport efficiency, in situ gas exchange and laboratory-based steady-state hydraulic conductance were examined for each leaf type. Both structure and physiology differed markedly between the two leaf types. Short-shoot leaves were thinner and had higher vein density, lower stomatal pore index, smaller bundle sheath extensions and lower hydraulic conductance than long-shoot leaves. Long shoots had lower xylem area:leaf area ratios than short shoots during leaf expansion, but this ratio was reversed at shoot maturity. Long-shoot leaves had higher rates of photosynthesis, stomatal conductance and transpiration than short-shoot leaves. We propose that structural differences between the two G. biloba leaf types reflect greater hydraulic limitation of long-shoot leaves during expansion. In turn, differences in physiological performance of short- and long-shoot leaves correspond to their distinct ontogeny and architecture.