Grace P. John

How Does Leaf Anatomy Influence Water Transport outside the Xylem

Anatomical data from diverse species, applied to a novel integrative model, elucidate the mechanistic basis of differences in water transport outside the xylem in leaves. Leaves are arguably the most complex and important physicobiological systems in the ecosphere. Yet, water transport outside the leaf xylem remains poorly understood, despite its impacts on stomatal function and photosynthesis. We applied anatomical measurements from 14 diverse species to a novel model of water flow in an areole (the smallest region bounded by minor veins) to predict the impact of anatomical variation across species on outside-xylem hydraulic conductance (Kox). Several predictions verified previous correlational studies: (1) vein length per unit area is the strongest anatomical determinant of Kox, due to effects on hydraulic pathlength and bundle sheath (BS) surface area; (2) palisade mesophyll remains well hydrated in hypostomatous species, which may benefit photosynthesis, (3) BS extensions enhance Kox; and (4) the upper and lower epidermis are hydraulically sequestered from one another despite their proximity. Our findings also provided novel insights: (5) the BS contributes a minority of outside-xylem resistance; (6) vapor transport contributes up to two-thirds of Kox; (7) Kox is strongly enhanced by the proximity of veins to lower epidermis; and (8) Kox is strongly influenced by spongy mesophyll anatomy, decreasing with protoplast size and increasing with airspace fraction and cell wall thickness. Correlations between anatomy and Kox across species sometimes diverged from predicted causal effects, demonstrating the need for integrative models to resolve causation. For example, (9) Kox was enhanced far more in heterobaric species than predicted by their having BS extensions. Our approach provides detailed insights into the role of anatomical variation in leaf function.

Allometry of cells and tissues within leaves

UNLABELLED PREMISE OF THE STUDY Allometric relationships among the dimensions of leaf cells, cell walls, and tissues, and whole-leaf thickness and area are likely to have key implications for leaf construction and function, but have remained virtually untested, despite the explosion of interest in allometric analysis of numerous plant properties at larger scales. • METHODS Using leaf transverse cross sections and light microscopy, we measured leaf dimensions, tissue thicknesses, mesophyll and xylem cell sizes, and cell wall thicknesses for 14 diverse angiosperm species of wet and dry habitats and tested hypothesized allometric relationships based on geometric scaling due to development and/or function. • KEY RESULTS We found strong novel allometries relating the dimensions of cells, cell walls, tissues, and gross leaf form. Cell sizes and cell wall thicknesses tended to scale isometrically across mesophyll tissues within the leaf, such that species with large cells or thick cell walls in one tissue had these also in the other tissues; however, leaf vein xylem conduit sizes were independent of those of other cell types. We also found strong geometric scaling of cell wall thicknesses with cell sizes throughout the mesophyll, but not in the leaf vein xylem. Further, leaf thickness scaled with cell sizes, cell wall thicknesses and the thicknesses of component mesophyll tissues, but leaf area was independent of anatomical traits across species. • CONCLUSIONS These novel allometries suggest design rules operating at the smallest scales of leaf construction and the possibility of applying these relationships to better characterizing the basis for differences among species in leaf form and functional traits.

Outside-xylem vulnerability, not xylem embolism, controls leaf hydraulic decline during dehydration

Changes in leaf outside-xylem properties drive leaf and whole-plant hydraulic decline with dehydration, protecting plants from catastrophic embolism in xylem conduits. Leaf hydraulic supply is crucial to maintaining open stomata for CO2 capture and plant growth. During drought-induced dehydration, the leaf hydraulic conductance (Kleaf) declines, which contributes to stomatal closure and, eventually, to leaf death. Previous studies have tended to attribute the decline of Kleaf to embolism in the leaf vein xylem. We visualized at high resolution and quantified experimentally the hydraulic vulnerability of xylem and outside-xylem pathways and modeled their respective influences on plant water transport. Evidence from all approaches indicated that the decline of Kleaf during dehydration arose first and foremost due to the vulnerability of outside-xylem tissues. In vivo x-ray microcomputed tomography of dehydrating leaves of four diverse angiosperm species showed that, at the turgor loss point, only small fractions of leaf vein xylem conduits were embolized, and substantial xylem embolism arose only under severe dehydration. Experiments on an expanded set of eight angiosperm species showed that outside-xylem hydraulic vulnerability explained 75% to 100% of Kleaf decline across the range of dehydration from mild water stress to beyond turgor loss point. Spatially explicit modeling of leaf water transport pointed to a role for reduced membrane conductivity consistent with published data for cells and tissues. Plant-scale modeling suggested that outside-xylem hydraulic vulnerability can protect the xylem from tensions that would induce embolism and disruption of water transport under mild to moderate soil and atmospheric droughts. These findings pinpoint outside-xylem tissues as a central locus for the control of leaf and plant water transport during progressive drought.

Leaf vein xylem conduit diameter influences susceptibility to embolism and hydraulic decline

Ecosystems worldwide are facing increasingly severe and prolonged droughts during which hydraulic failure from drought-induced embolism can lead to organ or whole plant death. Understanding the determinants of xylem failure across species is especially critical in leaves, the engines of plant growth. If the vulnerability segmentation hypothesis holds within leaves, higher order veins that are most terminal in the plant hydraulic system should be more susceptible to embolism to protect the rest of the water transport system. Increased vulnerability in the higher order veins would also be consistent with these experiencing the greatest tensions in the plant xylem network. To test this hypothesis, we combined X-ray micro-computed tomography imaging, hydraulic experiments, cross-sectional anatomy and 3D physiological modelling to investigate how embolisms spread throughout petioles and vein orders during leaf dehydration in relation to conduit dimensions. Decline of leaf xylem hydraulic conductance (Kx ) during dehydration was driven by embolism initiating in petioles and midribs across all species, and Kx vulnerability was strongly correlated with petiole and midrib conduit dimensions. Our simulations showed no significant impact of conduit collapse on Kx decline. We found xylem conduit dimensions play a major role in determining the susceptibility of the leaf water transport system during strong leaf dehydration.

The Sites of Evaporation within Leaves

Vapor transport within leaves influences tissue water potentials, requiring the reassessment of hypotheses related to the sites of evaporation, including that measured hydraulic and stomatal conductances are directly influenced by where evaporation occurs. The sites of evaporation within leaves are unknown, but they have drawn attention for decades due to their perceived implications for many factors, including patterns of leaf isotopic enrichment, the maintenance of mesophyll water status, stomatal regulation, and the interpretation of measured stomatal and leaf hydraulic conductances. We used a spatially explicit model of coupled water and heat transport outside the xylem, MOFLO 2.0, to map the distribution of net evaporation across leaf tissues in relation to anatomy and environmental parameters. Our results corroborate earlier predictions that most evaporation occurs from the epidermis at low light and moderate humidity but that the mesophyll contributes substantially when the leaf center is warmed by light absorption, and more so under high humidity. We also found that the bundle sheath provides a significant minority of evaporation (15% in darkness and 18% in high light), that the vertical center of amphistomatous leaves supports net condensation, and that vertical temperature gradients caused by light absorption vary over 10-fold across species, reaching 0.3°C. We show that several hypotheses that depend on the evaporating sites require revision in light of our findings, including that experimental measurements of stomatal and hydraulic conductances should be affected directly by changes in the location of the evaporating sites. We propose a new conceptual model that accounts for mixed-phase water transport outside the xylem. These conclusions have far-reaching implications for inferences in leaf hydraulics, gas exchange, water use, and isotope physiology.

Leaf mass per area is independent of vein length per area: avoiding pitfalls when modelling phenotypic integration (reply to Blonder et al. 2014)

Highlight text In a previous paper we clarified the roles of veins in determining leaf function. Here we contribute further data and address the concerns of Blonder et al. (2014), and provide guidance for phenotypic modelling.

ABA Accumulation in Dehydrating Leaves Is Associated with Decline in Cell Volume, Not Turgor Pressure

Reanalysis of published experimental data shows that in dehydrating leaves ABA accumulation is linked with reduction of cell volume rather than turgor, providing clues toward signaling mechanisms.

Embracing 3D Complexity in Leaf Carbon–Water Exchange

Leaves are a nexus for the exchange of water, carbon, and energy between terrestrial plants and the atmosphere. Research in recent decades has highlighted the critical importance of the underlying biophysical and anatomical determinants of CO2 and H2O transport, but a quantitative understanding of how detailed 3D leaf anatomy mediates within-leaf transport has been hindered by the lack of a consensus framework for analyzing or simulating transport and its spatial and temporal dynamics realistically, and by the difficulty of measuring within-leaf transport at the appropriate scales. We discuss how recent technological advancements now make a spatially explicit 3D leaf analysis possible, through new imaging and modeling tools that will allow us to address long-standing questions related to plant carbon-water exchange.