No time to rest: seasonal dynamics of non-structural carbohydrates in twigs of three Mediterranean tree species suggest year-round activity.

Perennial plants in temperate climates evolved short and long-term strategies to store and manage reserves in the form of non-structural carbohydrates (NSC; soluble sugars (SC) and starch (St)). NSC storage allows plants to survive seasonal periods of photosynthetic inactivity (dormancy). To study year-to-year seasonal patterns of trees' NSC dynamics that control phenology and yields, we established a large scale, multi-year study called the "Carbohydrate Observatory" using a citizen science approach with ~ 590 sites throughout the Central Valley of California. Monthly sampling tracked seasonal trends of starch and sugar levels in both xylem and phloem of twigs in Prunus dulcis, Pistacia vera and Juglans regia. Presented is the initial technical analysis of the first 3 years. With no exception, levels of reserves changed continuously throughout the year suggesting that even during dormancy, the average concentration of NSC, starch and sugars varies seasonally. In general, carbohydrate reserves are highest entering dormancy. During winter, NSCs slowly decrease to depletion during bloom time and remain low during summer until recovery near harvest. Starch is the major reserve compound in the wood of P. dulcis and P. vera while soluble sugars are the major reserves in J. regia. NSC content fluctuates throughout a season and significantly varies between years suggesting intrinsic and climatic effects on trees' energy reserves.

Hydraulic conductivity of red oak (Quercus rubra L.) leaf tissue does not respond to light.

The permeability of leaf tissue to water has been reported to increase under illumination, a response reputed to involve aquaporins. We studied this 'light response' in red oak (Quercus rubra L.), the species in which the phenomenon was first detected during measurements of leaf hydraulic conductance with the high-pressure flow meter (HPFM). In our HPFM measurements, we found that pre-conditioning leaves in darkness was not sufficient to bring them to their minimum conductance, which was attained only after an hour of submersion and pressurization. However, pre-conditioning leaves under anoxic conditions resulted in an immediate reduction in conductance. Leaves light- and dark-acclimated while on the tree showed no differences in the time course of HPFM measurement under illumination. We also studied the effect of light level and anoxia on rehydration kinetics, finding that anoxia slowed rehydration, but light had no effect either in the lab (rehydration under low light, high humidity) or on the tree (acclimation under high light, 10 min of dark prior to rehydration). We conclude that the declines in conductance observed in the HPFM must involve a resistance downstream of the extracellular air space, and that in red oak the hydraulic conductivity of leaf tissue is insensitive to light.

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 (L(stem)), 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.

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.

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 (delta), 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.

In vivo observation of cavitation and embolism repair using magnetic resonance imaging.

Magnetic resonance imaging (MRI) was used to noninvasively monitor the status of individual xylem vessels in the stem of an intact, transpiring grape (Vitis vinifera) plant over a period of approximately 40 h. Proton density-weighted MRI was used to visualize the distribution of mobile water in the stem and individual xylem vessels were scored as either water or gas filled (i.e. embolized). The number of water-filled vessels decreased during the first 24 h of the experiment, indicating that approximately 10 vessels had cavitated during this time. Leaf water potentials decreased from -1.25 to -2.1 MPa during the same period. Watering increased leaf water potentials to -0.25 MPa and prevented any further cavitation. Refilling of xylem vessels occurred as soon as the lights were switched off, with the majority of vessels becoming refilled with water during the first 2 to 3 h in darkness. These measurements demonstrate that MRI can be used to monitor the functional status of individual xylem vessels, providing the first method to study the process of cavitation and embolism repair in intact plants.

Hydraulic properties of individual xylem vessels of Fraxinus americana.

Studies of the hydraulic properties of xylem vessels have been limited to measurements of whole plant or whole stem segments. This approach allows the longitudinal transport properties of the ensemble of vessels within a stem to be determined, but provides little information on radial transport. Here the xylem of Fraxinus americana L. has been examined using a new method that allows the transport properties of individual vessels to be examined. One goal of this study was to quantify transport parameters relevant to embolism repair. The longitudinal conductivity of vessel segments open at both ends (i.e. no end walls) agreed with values predicted by the Poiseuille equation. Radial specific conductance (conductance per unit area) was approximately six orders of magnitude lower than the longitudinal conductance of the vessel segment normalized by the cross-sectional area of the vessel lumen. There was a step increase in the radial specific conductance of previously gas-filled vessels when the delivery pressure exceeded 0.4 MPa. This is consistent with the idea that positive pressure, required for embolism repair, can be compartmentalized within a vessel if the bordered pit chambers are gas-filled. The diffusion coefficient for the movement of gas from a pressurized air-filled vessel was of the same order of magnitude as that for air diffusing through water (1.95 e(-9) m(2) s(-1)). Estimates of the time needed to displace all of the gas from an air-filled vessel were in the order of 20 min, suggesting that gas removal may not be a major limitation in embolism repair.

Hydrogel control of xylem hydraulic resistance in plants.

Increasing concentrations of ions flowing through the xylem of plants produce rapid, substantial, and reversible decreases in hydraulic resistance. Changes in hydraulic resistance in response to solution ion concentration, pH, and nonpolar solvents are consistent with this process being mediated by hydrogels. The effect is localized to intervessel bordered pits, suggesting that microchannels in the pit membranes are altered by the swelling and deswelling of pectins, which are known hydrogels. The existence of an ion-mediated response breaks the long-held paradigm of the xylem as a system of inert pipes and suggests a mechanism by which plants may regulate their internal flow regime.

Bordered pit structure and vessel wall surface properties. Implications for embolism repair.

The idea that embolized xylem vessels can be refilled while adjacent vessels remain under tension is difficult to accept if the cavitated vessels remain hydraulically connected to vessels under tension. A mechanism by which embolized conduits could be hydraulically isolated from adjacent conduits requires the existence of a non-zero contact angle and a flared opening into the bordered pit chamber such that a convex air-water interface forms at the entrance into the pit chamber. We measured the contact angle and pit chamber geometry for six species. The contact angle measured in the vessel lumen ranged between 42 degrees to 55 degrees, whereas the opening into the pit chamber ranged between 144 degrees and 157 degrees. If the surface properties within the pit chamber are similar to those in the lumen, a convex meniscus will form at the flared opening into the pit chamber. The maximum pressure difference between water in the lumen and gas in the pit chamber that could be stabilized by this interface was calculated to be within the range of 0.07 to 0.30 MPa.

Stomatal plugs of Drimys winteri (Winteraceae) protect leaves from mist but not drought.

Two outstanding features of the flowering plant family Winteraceae are the occlusion of their stomatal pores by cutin plugs and the absence of water-conducting xylem vessels. An adaptive relationship between these two unusual features has been suggested whereby stomatal plugs restrict gas exchange to compensate for the presumed poor conductivity of their vesselless wood. This hypothesized connection fueled evolutionary arguments that the vesselless condition is ancestral in angiosperms. Here we show that in Drimys winteri, a tree common to wet forests, these stomatal occlusions pose only a small fixed resistance to water loss. In addition, they modify the humidity response of guard cells such that under high evaporative demand, leaves with plugs lose water at a faster rate than leaves from which the plugs have been experimentally removed. Instead of being adaptations for drought, we present evidence that these cuticular structures function to maintain photosynthetic activity under conditions of excess water on the leaf surface. Stomatal plugs decrease leaf wettability by preventing the formation of a continuous water film that would impede diffusion of CO2 into the leaf. Misting of leaves had no effect on photosynthetic rate of leaves with plugs, but resulted in a marked decrease ( approximately 40%) in leaves from which the plugs had been removed. These findings do not support a functional association between stomatal plugs and hydraulic competence and provide a new perspective on debates surrounding the evolution of vessels in angiosperms.