Publications

2016
Knoblauch, Michael, Jan Knoblauch, Daniel L Mullendore, Jessica A Savage, Benjamin A Babst, Sierra D Beecher, Adam C Dodgen, Kaare H Jensen, and Noel Michele Holbrook. 2016. “Testing the Münch hypothesis of long distance phloem transport in plants.” eLife 5.Abstract

Long distance transport in plants occurs in sieve tubes of the phloem. The pressure flow hypothesis introduced by Ernst Münch in 1930 describes a mechanism of osmotically generated pressure differentials that are supposed to drive the movement of sugars and other solutes in the phloem, but this hypothesis has long faced major challenges. The key issue is whether the conductance of sieve tubes, including sieve plate pores, is sufficient to allow pressure flow. We show that with increasing distance between source and sink, sieve tube conductivity and turgor increases dramatically inIpomoea nil. Our results provide strong support for the Münch hypothesis, while providing new tools for the investigation of one of the least understood plant tissues.

Munch paper
2015
Rolland, V., D.M. Bergstrom, T. Lenne, G. Bryant, H.Chen, J. Wolfe, M. N Holbrook, D. E. Stanton, and M.C. Ball. 2015. “Easy Come, Easy Go: Capillary Forces Enable Rapid Refilling of Embolized Primary Xylem Vessels.” Plant Physiology American Society of Plant Biologists , 1636-1647.Abstract

Protoxylem plays an important role in the hydraulic function of vascular systems of both herbaceous and woody plants, but relatively little is known about the processes underlying the maintenance of protoxylem function in long-lived tissues. In this study, embolism repair was investigated in relation to xylem structure in two cushion plant species, Azorella macquariensis and Colobanthus muscoides, in which vascular water transport depends on protoxylem. Their protoxylem vessels consisted of a primary wall with helical thickenings that effectively formed a pit channel, with the primary wall being the pit channel membrane. Stem protoxylem was organized such that the pit channel membranes connected vessels with paratracheal parenchyma or other protoxylem vessels and were not exposed directly to air spaces. Embolism was experimentally induced in excised vascular tissue and detached shoots by exposing them briefly to air. When water was resupplied, embolized vessels refilled within tens of seconds (excised tissue) to a few minutes (detached shoots) with water sourced from either adjacent parenchyma or water-filled vessels. Refilling occurred in two phases: (1) water refilled xylem pit channels, simplifying bubble shape to a rod with two menisci; and (2) the bubble contracted as the resorption front advanced, dissolving air along the way. Physical properties of the protoxylem vessels (namely pit channel membrane porosity, hydrophilic walls, vessel dimensions, and helical thickenings) promoted rapid refilling of embolized conduits independent of root pressure. These results have implications for the maintenance of vascular function in both herbaceous and woody species, because protoxylem plays a major role in the hydraulic systems of leaves, elongating stems, and roots.

Dietterich, Lee H., Antonella Zanobetti, Itai Kloog, Peter Huybers, Andrew D.B. Leakey, Arnold J. Bloom, Eli Carlisle, et al. 2015. “Impacts of elevated atmospheric CO2 on nutrient content of important food crops.” Scientific Data, no. 150036.
Stein, Ofer, Hila Damari-Weissler, Francesca Secchi, Shimon Rachamilevitch, Marcelo A. German, Yelena Yeselson, Rachel Amir, et al. 2015. “The tomato plastidic fructokinase SIFRK3 plays a role in xylem development.” New Phytologist , 1484-1495.
Watkins, James E., Amber C. Churchill, and Noel Michele Holbrook. 2015. “A site for sori: Ecophysiology of fertile–sterile leaf dimorphy in ferns.” American journal of botany 103 (5): 845-855.Abstract

PREMISE OF THE STUDY: Reproduction often requires significant investment and can move resources away from growth and maintenance; maintaining a balance between reproduction and growth can involve trade-offs. Extreme functional specialization has separated reproduction and photosynthesis in most seed plants, yet ferns use the laminar surface of their fronds for both reproduction and photosynthesis. This dual function selects for a variety of frond morphologies that range from no specialization (monomorphy) to extreme dimorphy between fertile and sterile fronds (holodimorphy). Here we examined the ecological and physiological consequences of variation in frond dimorphy in ferns, evaluated reproductive trade-offs across a dimorphy gradient, and speculate on factors controlling the occurrence of holodimorphy.

METHODS: Ecophysiological measurements of photosynthetic rate, water potential, hydraulic conductivity, and gross morphological comparisons of frond area and angle were used to evaluate differences between fertile and sterile fronds. We examined three temperate and three tropical fern species that vary in degree of fertile–sterile dimorphy.

KEY RESULTS: Holodimorphic species produced fewer fertile fronds, which had significantly higher respiratory rates than in sterile fronds on the same plant or in any frond produced on monomorphic species; hemidimorphic species were frequently intermediate. We found no differences in vulnerability to cavitation between fertile and sterile fronds. In dimorphic species, fertile fronds had higher (less negative) water potential and lower stipe hydraulic conductivity relative than in sterile fronds.

CONCLUSIONS: Fertile–sterile dimorphy in ferns appears to come at considerable carbon cost in holodimorohic species. It is possible that the relative costs of this reproductive system are offset by increased spore dispersal, yet such trade-offs require further exploration.

Nathaniel D. Mueller,, Ethan E. Butler, Karen A. McKinnon, Andrew Rhines, Marin Tingley, N. Michele Holbrook, and Peter Huybers. 2015. “Cooling of US Midwest summer temperature extremes from cropland intensification.” Nature Climate Change . Publisher's Version
Ronellenfitsch, H, J Liesche, KH Jensen, NM Holbrook, A Schulz, and E Katifori. 2015. “Scaling of phloem structure and optimality of photoassimilate transport in conifer needles.” Proc Biol Sci 282: 20141863.Abstract

The phloem vascular system facilitates transport of energy-rich sugar and signalling molecules in plants, thus permitting long-range communication within the organism and growth of non-photosynthesizing organs such as roots and fruits. The flow is driven by osmotic pressure, generated by differences in sugar concentration between distal parts of the plant. The phloem is an intricate distribution system, and many questions about its regulation and structural diversity remain unanswered. Here, we investigate the phloem structure in the simplest possible geometry: a linear leaf, found, for example, in the needles of conifer trees. We measure the phloem structure in four tree species representing a diverse set of habitats and needle sizes, from 1 (Picea omorika) to 35 cm (Pinus palustris). We show that the phloem shares common traits across these four species and find that the size of its conductive elements obeys a power law. We present a minimal model that accounts for these common traits and takes into account the transport strategy and natural constraints. This minimal model predicts a power law phloem distribution consistent with transport energy minimization, suggesting that energetics are more important than translocation speed at the leaf level.

2014
Rockwell, Fulton E., N. Michele Holbrook, and Abraham D. Stroock. 2014. “Leaf hydraulics I: Scaling transport properties from single cells to tissue.” Journal of Theoretical Biology 340: 251-266.Abstract

 

In leaf tissues, water may move through the symplast or apoplast as a liquid, or through the airspace as vapor, but the dominant path remains in dispute. This is due, in part, to a lack of models that describe these three pathways in terms of experimental variables. We show that, in plant water relations theory, the use of a hydraulic capacity in a manner analogous to a thermal capacity, though it ignores mechanical interactions between cells, is consistent with a special case of the more general continuum mechanical theory of linear poroelasticity. The resulting heat equation form affords a great deal of analytical simplicity at a minimal cost: we estimate an expected error of less than 12%, compared to the full set of equations governing linear poroelastic behavior. We next consider the case for local equilibrium between protoplasts, their cell walls, and adjacent air spaces during isothermal hydration transients to determine how accurately simple volume averaging of material properties (a compositemodel) describes the hydraulic properties of leaf tissue. Based on typical hydraulic parameters for individual cells, we find that a composite description for tissues composed of thin walled cells with air spaces of similar size to the cells, as in photosynthetic tissues, is a reasonable preliminary assumption. We also expect isothermal transport in such cells to be dominated by the aquaporin-mediated cell-to-cell path. In the non- isothermal case, information on the magnitude of the thermal gradients is required to assess the dominant phase of water transport, liquid or vapor.

Stroock, A. D., Vinay V. Pagay, M. A. Zwieniecki, and N. M. Holbrook. 2014. “The Physicochemical Hydrodynamics of Vascular Plants.” Annual Review of Fluid Mechanics 46: 615-642.Abstract

 

Plants live dangerously, but gracefully. To remain hydrated, they exploit liquid water in the thermodynamically metastable state of negative pressure, similar to a rope under tension. This tension allows them to pull water out of the soil and up to their leaves. When this liquid rope breaks, owing to cavitation, they catch the ends to keep it from unraveling and then bind it back together. In parallel, they operate a second vascular system for the circulation of metabolites though their tissues, this time with positive pressures and flow that passes from leaf to root. In this article, we review the current state of understanding of water management in plants with an emphasis on the rich coupling of transport phenomena, thermodynamics, and active biological processes. We discuss efforts to replicate plant function in synthetic systems and point to opportunities for physical scientists and engineers to benefit from and contribute to the study of plants.

Mello, F.C., C.E.P. Cerri, C.A. Davies, N. M. Holbrook, K. Paustian, S.M.F. Maia, M.V. Galdos, M. Bernoux, and C.C. Cerri. 2014. “Payback time for soil carbon and sugar-cane ethanol.” nature climate change, 605-609.
Rockwell, FE, JK Wheeler, and NM Holbrook. 2014. “Cavitation and its discontents: opportunities for resolving current controversies.” Plant Physiol 164: 1649-60.Abstract

Cavitation has long been recognized as a key constraint on the structure and functional integrity of the xylem. Yet, recent results call into question how well we understand cavitation in plants. Here, we consider embolism formation in angiosperms at two scales. The first focuses on how air-seeding occurs at the level of pit membranes, raising the question of whether capillary failure is an appropriate physical model. The second addresses methodological uncertainties that affect our ability to infer the formation of embolism and its reversal in plant stems. Overall, our goal is to open up fresh perspectives on the structure-function relationships of xylem.

Rockwell, FE, NM Holbrook, and AD Stroock. 2014. “The competition between liquid and vapor transport in transpiring leaves.” Plant Physiol 164: 1741-58.Abstract

In leaves, the transpirational flux of water exits the veins as liquid and travels toward the stomata in both the vapor and liquid phases before exiting the leaf as vapor. Yet, whether most of the evaporation occurs from the vascular bundles (perivascular), from the photosynthetic mesophyll cells, or within the vicinity of the stomatal pore (peristomatal) remains in dispute. Here, a one-dimensional model of the competition between liquid and vapor transport is developed from the perspective of nonisothermal coupled heat and water molecule transport in a composite medium of airspace and cells. An analytical solution to the model is found in terms of the energy and transpirational fluxes from the leaf surfaces and the absorbed solar energy load, leading to mathematical expressions for the proportions of evaporation accounted for by the vascular, mesophyll, and epidermal regions. The distribution of evaporation in a given leaf is predicted to be variable, changing with the local environment, and to range from dominantly perivascular to dominantly peristomatal depending on internal leaf architecture, with mesophyll evaporation a subordinate component. Using mature red oak (Quercus rubra) trees, we show that the model can be solved for a specific instance of a transpiring leaf by combining gas-exchange data, anatomical measurements, and hydraulic experiments. We also investigate the effect of radiation load on the control of transpiration, the potential for condensation on the inside of an epidermis, and the impact of vapor transport on the hydraulic efficiency of leaf tissue outside the xylem.

Myers, SS, A Zanobetti, I Kloog, P Huybers, AD Leakey, AJ Bloom, E Carlisle, et al. 2014. “Increasing CO2 threatens human nutrition.” Nature 510: 139-42.Abstract

Dietary deficiencies of zinc and iron are a substantial global public health problem. An estimated two billion people suffer these deficiencies, causing a loss of 63 million life-years annually. Most of these people depend on C3 grains and legumes as their primary dietary source of zinc and iron. Here we report that C3 grains and legumes have lower concentrations of zinc and iron when grown under field conditions at the elevated atmospheric CO2 concentration predicted for the middle of this century. C3 crops other than legumes also have lower concentrations of protein, whereas C4 crops seem to be less affected. Differences between cultivars of a single crop suggest that breeding for decreased sensitivity to atmospheric CO2 concentration could partly address these new challenges to global health.

Rockwell, FE, NM Holbrook, and AD Stroock. 2014. “Leaf hydraulics II: vascularized tissues.” J Theor Biol 340: 267-84.Abstract

Current models of leaf hydration employ an Ohm's law analogy of the leaf as an ideal capacitor, neglecting the resistance to flow between cells, or treat the leaf as a plane sheet with a source of water at fixed potential filling the mid-plane, neglecting the discrete placement of veins as well as their resistance. We develop a model of leaf hydration that considers the average conductance of the vascular network to a representative areole (region bounded by the vascular network), and represent the volume of tissue within the areole as a poroelastic composite of cells and air spaces. Solutions to the 3D flow problem are found by numerical simulation, and these results are then compared to 1D models with exact solutions for a range of leaf geometries, based on a survey of temperate woody plants. We then show that the hydration times given by these solutions are well approximated by a sum of the ideal capacitor and plane sheet times, representing the time for transport through the vasculature and tissue respectively. We then develop scaling factors relating this approximate solution to the 3D model, and examine the dependence of these scaling factors on leaf geometry. Finally, we apply a similar strategy to reduce the dimensions of the steady state problem, in the context of peristomatal transpiration, and consider the relation of transpirational gradients to equilibrium leaf water potential measurements.

Savage, JA, DF Haines, and NM Holbrook. 2014. “The making of giant pumpkins: how selective breeding changed the phloem of Cucurbita maxima from source to sink.” Plant Cell Environ.Abstract

Despite the success of breeding programmes focused on increasing fruit size, relatively little is known about the anatomical and physiological changes required to increase reproductive allocation. To address this gap in knowledge, we compared fruit/ovary anatomy, vascular structure and phloem transport of two varieties of giant pumpkins, and their smaller fruited progenitor under controlled environmental conditions. We also modelled carbon transport into the fruit of competitively grown plants using data collected in the field. There was no evidence that changes in leaf area or photosynthetic capacity impacted fruit size. Instead, giant varieties differed in their ovary morphology and contained more phloem on a cross-sectional area basis in their petioles and pedicels than the ancestral variety. These results suggest that sink activity is important in determining fruit size and that giant pumpkins have an enhanced capacity to transport carbon. The strong connection observed between carbon fixation, phloem structure and fruit growth in field-grown plants indicates that breeding for large fruit has led to changes throughout the carbon transport system that could have important implications for how we think about phloem transport velocity and carbon allocation.

Carroll, NJ, KH Jensen, S Parsa, NM Holbrook, and DA Weitz. 2014. “Measurement of flow velocity and inference of liquid viscosity in a microfluidic channel by fluorescence photobleaching.” Langmuir 30: 4868-74.Abstract

We present a simple, noninvasive method for simultaneous measurement of flow velocity and inference of liquid viscosity in a microfluidic channel. We track the dynamics of a sharp front of photobleached fluorescent dye using a confocal microscope and measure the intensity at a single point downstream of the initial front position. We fit an exact solution of the advection diffusion equation to the fluorescence intensity recovery curve to determine the average flow velocity and the diffusion coefficient of the tracer dye. The dye diffusivity is correlated to solute concentration to infer rheological properties of the liquid. This technique provides a simple method for simultaneous elucidation of flow velocity and liquid viscosity in microchannels.

Neumann, RB, ZG Cardon, J Teshera-Levye, FE Rockwell, MA Zwieniecki, and NM Holbrook. 2014. “Modelled hydraulic redistribution by sunflower (Helianthus annuus L.) matches observed data only after including night-time transpiration.” Plant Cell Environ 37: 899-910.Abstract

The movement of water from moist to dry soil layers through the root systems of plants, referred to as hydraulic redistribution (HR), occurs throughout the world and is thought to influence carbon and water budgets and ecosystem functioning. The realized hydrologic, biogeochemical and ecological consequences of HR depend on the amount of redistributed water, whereas the ability to assess these impacts requires models that correctly capture HR magnitude and timing. Using several soil types and two ecotypes of sunflower (Helianthus annuus L.) in split-pot experiments, we examined how well the widely used HR modelling formulation developed by Ryel et al. matched experimental determination of HR across a range of water potential driving gradients. H. annuus carries out extensive night-time transpiration, and although over the last decade it has become more widely recognized that night-time transpiration occurs in multiple species and many ecosystems, the original Ryel et al. formulation does not include the effect of night-time transpiration on HR. We developed and added a representation of night-time transpiration into the formulation, and only then was the model able to capture the dynamics and magnitude of HR we observed as soils dried and night-time stomatal behaviour changed, both influencing HR.

Kelly, G, N Sade, Z Attia, F Secchi, M Zwieniecki, NM Holbrook, A Levi, V Alchanatis, M Moshelion, and D Granot. 2014. “Relationship between hexokinase and the aquaporin PIP1 in the regulation of photosynthesis and plant growth.” PLoS One 9: e87888.Abstract

Increased expression of the aquaporin NtAQP1, which is known to function as a plasmalemma channel for CO(2) and water, increases the rate of both photosynthesis and transpiration. In contrast, increased expression of Arabidopsis hexokinase1 (AtHXK1), a dual-function enzyme that mediates sugar sensing, decreases the expression of photosynthetic genes and the rate of transpiration and inhibits growth. Here, we show that AtHXK1 also decreases root and stem hydraulic conductivity and leaf mesophyll CO(2) conductance (g(m)). Due to their opposite effects on plant development and physiology, we examined the relationship between NtAQP1 and AtHXK1 at the whole-plant level using transgenic tomato plants expressing both genes simultaneously. NtAQP1 significantly improved growth and increased the transpiration rates of AtHXK1-expressing plants. Reciprocal grafting experiments indicated that this complementation occurs when both genes are expressed simultaneously in the shoot. Yet, NtAQP1 had only a marginal effect on the hydraulic conductivity of the double-transgenic plants, suggesting that the complementary effect of NtAQP1 is unrelated to shoot water transport. Rather, NtAQP1 significantly increased leaf mesophyll CO(2) conductance and enhanced the rate of photosynthesis, suggesting that NtAQP1 facilitated the growth of the double-transgenic plants by enhancing mesophyll conductance of CO(2).

Zhang, YJ, FE Rockwell, JK Wheeler, and NM Holbrook. 2014. “Reversible Deformation of Transfusion Tracheids in Taxus baccata Is Associated with a Reversible Decrease in Leaf Hydraulic Conductance.” Plant Physiol 165: 1557-1565.Abstract

Declines in leaf hydraulic conductance (Kleaf) with increasing water stress have been attributed to cavitation of the leaf xylem. However, in the leaves of conifers, the reversible collapse of transfusion tracheids may provide an alternative explanation. Using Taxus baccata, a conifer species without resin, we developed a modified rehydration technique that allows the separation of declines in Kleaf into two components: one reversible and one irreversible upon relaxation of water potential to -1 MPa. We surveyed leaves at a range of water potentials for evidence of cavitation using cryo-scanning electron microscopy and quantified dehydration-induced structural changes in transfusion tracheids by cryo-fluorescence microscopy. Irreversible declines in Kleaf did not occur until leaf water potentials were more negative than -3 MPa. Declines in Kleaf between -2 and -3 MPa were reversible and accompanied by the collapse of transfusion tracheids, as evidenced by cryo-fluorescence microscopy. Based on cryo-scanning electron microscopy, cavitation of either transfusion or xylem tracheids did not contribute to declines in Kleaf in the reversible range. Moreover, the deformation of transfusion tracheids was quickly reversible, thus acting as a circuit breaker regulating the flux of water through the leaf vasculature. As transfusion tissue is present in all gymnosperms, the reversible collapse of transfusion tracheids may be a general mechanism in this group for the protection of leaf xylem from excessive loads generated in the living leaf tissue.

Zhang, YJ, NM Holbrook, and KF Cao. 2014. “Seasonal dynamics in photosynthesis of woody plants at the northern limit of Asian tropics: potential role of fog in maintaining tropical rainforests and agriculture in Southwest China.” Tree Physiol 34: 1069-78.Abstract

The lowland tropical rainforests in Xishuangbanna, Southwest (SW) China, mark the northern limit of Asian tropics. Fog has been hypothesized to play a role in maintaining rainforests and tropical crop production in this region, but the physiological mechanism has not been studied. The goals of this study were to characterize the seasonal dynamics in photosynthesis and to assess the potential for fog to mitigate chilling-induced photodamage for tropical trees and crops in Xishuangbanna. We measured seasonal dynamics in light-saturated net photosynthetic rate (Aa), stomatal conductance (gs), intercellular CO2 concentration, quantum yield of Photosystem II (Fv/Fm) and maximum P700 changes (Pm; indicates the amount of active PSI complex), as well as chilling resistance and fog (light/shading) effects on low temperature-induced decline in Fv/Fm and Pm for native tree and introduced lower latitude tree or woody shrub species grown in a tropical botanical garden. Despite significant decreases in Aa, gs, Pm and Fv/Fm, most species maintained considerably high Aa during the cool season (2.51-14.6 mumol m(-2) s(-1)). Shaded leaves exposed to seasonal low temperatures had higher Fv/Fm than sun-exposed leaves in the cool season. All species could tolerate 1.4 degrees C in the dark, whereas a combined treatment of low temperature and high light caused a distinctly faster decline in Pm and Fv/Fm compared with low temperature treatment alone. Because fog persistence avoids or shortens the duration of high light condition in the morning when the temperatures are still low, our results provide support for the hypothesis that fog reduces chilling damage to tropical plants in this region and thus plays a role in maintaining tropical rainforests and agriculture in SW China.

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