Is desiccation tolerance and avoidance reflected in xylem and phloem anatomy of two coexisting arid-zone coniferous trees?

Plants close their stomata during drought to avoid excessive water loss, but species differ in respect to the drought severity at which stomata close. The stomatal closure point is related to xylem anatomy and vulnerability to embolism, but it also has implications for phloem transport and possibly phloem anatomy to allow sugar transport at low water potentials. Desiccation-tolerant plants that close their stomata at severe drought should have smaller xylem conduits and/or fewer and smaller interconduit pits to reduce vulnerability to embolism but more phloem tissue and larger phloem conduits compared with plants that avoid desiccation. These anatomical differences could be expected to increase in response to long-term reduction in precipitation. To test these hypotheses, we used tridimensional synchroton X-ray microtomograph and light microscope imaging of combined xylem and phloem tissues of 2 coniferous species: one-seed juniper (Juniperus monosperma) and piñon pine (Pinus edulis) subjected to precipitation manipulation treatments. These species show different xylem vulnerability to embolism, contrasting desiccation tolerance, and stomatal closure points. Our results support the hypothesis that desiccation tolerant plants require higher phloem transport capacity than desiccation avoiding plants, but this can be gained through various anatomical adaptations in addition to changing conduit or tissue size.

A multi-species synthesis of physiological mechanisms in drought-induced tree mortality.

Widespread tree mortality associated with drought has been observed on all forested continents and global change is expected to exacerbate vegetation vulnerability. Forest mortality has implications for future biosphere-atmosphere interactions of carbon, water and energy balance, and is poorly represented in dynamic vegetation models. Reducing uncertainty requires improved mortality projections founded on robust physiological processes. However, the proposed mechanisms of drought-induced mortality, including hydraulic failure and carbon starvation, are unresolved. A growing number of empirical studies have investigated these mechanisms, but data have not been consistently analysed across species and biomes using a standardized physiological framework. Here, we show that xylem hydraulic failure was ubiquitous across multiple tree taxa at drought-induced mortality. All species assessed had 60% or higher loss of xylem hydraulic conductivity, consistent with proposed theoretical and modelled survival thresholds. We found diverse responses in non-structural carbohydrate reserves at mortality, indicating that evidence supporting carbon starvation was not universal. Reduced non-structural carbohydrates were more common for gymnosperms than angiosperms, associated with xylem hydraulic vulnerability, and may have a role in reducing hydraulic function. Our finding that hydraulic failure at drought-induced mortality was persistent across species indicates that substantial improvement in vegetation modelling can be achieved using thresholds in hydraulic function.

Prolonged experimental drought reduces plant hydraulic conductance and transpiration and increases mortality in a piñon-juniper woodland.

Plant hydraulic conductance (k s) is a critical control on whole-plant water use and carbon uptake and, during drought, influences whether plants survive or die. To assess long-term physiological and hydraulic responses of mature trees to water availability, we manipulated ecosystem-scale water availability from 2007 to 2013 in a piñon pine (Pinus edulis) and juniper (Juniperus monosperma) woodland. We examined the relationship between k s and subsequent mortality using more than 5 years of physiological observations, and the subsequent impact of reduced hydraulic function and mortality on total woody canopy transpiration (E C) and conductance (G C). For both species, we observed significant reductions in plant transpiration (E) and k s under experimentally imposed drought. Conversely, supplemental water additions increased E and k s in both species. Interestingly, both species exhibited similar declines in k s under the imposed drought conditions, despite their differing stomatal responses and mortality patterns during drought. Reduced whole-plant k s also reduced carbon assimilation in both species, as leaf-level stomatal conductance (g s) and net photosynthesis (A n) declined strongly with decreasing k s. Finally, we observed that chronically low whole-plant k s was associated with greater canopy dieback and mortality for both piñon and juniper and that subsequent reductions in woody canopy biomass due to mortality had a significant impact on both daily and annual canopy E C and G C. Our data indicate that significant reductions in k s precede drought-related tree mortality events in this system, and the consequence is a significant reduction in canopy gas exchange and carbon fixation. Our results suggest that reductions in productivity and woody plant cover in piñon-juniper woodlands can be expected due to reduced plant hydraulic conductance and increased mortality of both piñon pine and juniper under anticipated future conditions of more frequent and persistent regional drought in the southwestern United States.

Integrating ecophysiology and forest landscape models to improve projections of drought effects under climate change.

Fundamental drivers of ecosystem processes such as temperature and precipitation are rapidly changing and creating novel environmental conditions. Forest landscape models (FLM) are used by managers and policy-makers to make projections of future ecosystem dynamics under alternative management or policy options, but the links between the fundamental drivers and projected responses are weak and indirect, limiting their reliability for projecting the impacts of climate change. We developed and tested a relatively mechanistic method to simulate the effects of changing precipitation on species competition within the LANDIS-II FLM. Using data from a field precipitation manipulation experiment in a piñon pine (Pinus edulis) and juniper (Juniperus monosperma) ecosystem in New Mexico (USA), we calibrated our model to measurements from ambient control plots and tested predictions under the drought and irrigation treatments against empirical measurements. The model successfully predicted behavior of physiological variables under the treatments. Discrepancies between model output and empirical data occurred when the monthly time step of the model failed to capture the short-term dynamics of the ecosystem as recorded by instantaneous field measurements. We applied the model to heuristically assess the effect of alternative climate scenarios on the piñon-juniper ecosystem and found that warmer and drier climate reduced productivity and increased the risk of drought-induced mortality, especially for piñon. We concluded that the direct links between fundamental drivers and growth rates in our model hold great promise to improve our understanding of ecosystem processes under climate change and improve management decisions because of its greater reliance on first principles.

Carbohydrate dynamics and mortality in a piñon-juniper woodland under three future precipitation scenarios.

Drought-induced forest mortality is an increasing global problem with wide-ranging consequences, yet mortality mechanisms remain poorly understood. Depletion of non-structural carbohydrate (NSC) stores has been implicated as an important mechanism in drought-induced mortality, but experimental field tests are rare. We used an ecosystem-scale precipitation manipulation experiment to evaluate leaf and twig NSC dynamics of two co-occurring conifers that differ in patterns of stomatal regulation of water loss and recent mortality: the relatively desiccation-avoiding piñon pine (Pinus edulis) and the relatively desiccation-tolerant one-seed juniper (Juniperus monosperma). Piñon pine experienced 72% mortality after 13-25 months of experimental drought and juniper experienced 20% mortality after 32-47 months. Juniper maintained three times more NSC in the foliage than twigs, and converted NSC to glucose and fructose under drought, consistent with osmoregulation requirements to maintain higher stomatal conductance during drought than piñon. Despite these species differences, experimental drought caused decreased leaf starch content in dying trees of both species (P < 0.001). Average dry-season leaf starch content was also a good predictor of drought-survival time for both species (R(2) = 0.93). These results, along with observations of drought-induced reductions to photosynthesis and growth, support carbon limitation as an important process during mortality of these two conifer species.

Constraining 3-PG with a new δ13C submodel: a test using the δ13C of tree rings.

A semi-mechanistic forest growth model, 3-PG (Physiological Principles Predicting Growth), was extended to calculate δ(13)C in tree rings. The δ(13)C estimates were based on the model's existing description of carbon assimilation and canopy conductance. The model was tested in two ~80-year-old natural stands of Abies grandis (grand fir) in northern Idaho. We used as many independent measurements as possible to parameterize the model. Measured parameters included quantum yield, specific leaf area, soil water content and litterfall rate. Predictions were compared with measurements of transpiration by sap flux, stem biomass, tree diameter growth, leaf area index and δ(13)C. Sensitivity analysis showed that the model's predictions of δ(13)C were sensitive to key parameters controlling carbon assimilation and canopy conductance, which would have allowed it to fail had the model been parameterized or programmed incorrectly. Instead, the simulated δ(13)C of tree rings was no different from measurements (P > 0.05). The δ(13)C submodel provides a convenient means of constraining parameter space and avoiding model artefacts. This δ(13)C test may be applied to any forest growth model that includes realistic simulations of carbon assimilation and transpiration.

How do trees die? A test of the hydraulic failure and carbon starvation hypotheses.

Despite decades of research on plant drought tolerance, the physiological mechanisms by which trees succumb to drought are still under debate. We report results from an experiment designed to separate and test the current leading hypotheses of tree mortality. We show that piñon pine (Pinus edulis) trees can die of both hydraulic failure and carbon starvation, and that during drought, the loss of conductivity and carbohydrate reserves can also co-occur. Hydraulic constraints on plant carbohydrate use determined survival time: turgor loss in the phloem limited access to carbohydrate reserves, but hydraulic control of respiration prolonged survival. Our data also demonstrate that hydraulic failure may be associated with loss of adequate tissue carbohydrate content required for osmoregulation, which then promotes failure to maintain hydraulic integrity.

Evaluating theories of drought-induced vegetation mortality using a multimodel-experiment framework.

SUMMARY: Model-data comparisons of plant physiological processes provide an understanding of mechanisms underlying vegetation responses to climate. We simulated the physiology of a piñon pine-juniper woodland (Pinus edulis-Juniperus monosperma) that experienced mortality during a 5 yr precipitation-reduction experiment, allowing a framework with which to examine our knowledge of drought-induced tree mortality. We used six models designed for scales ranging from individual plants to a global level, all containing state-of-the-art representations of the internal hydraulic and carbohydrate dynamics of woody plants. Despite the large range of model structures, tuning, and parameterization employed, all simulations predicted hydraulic failure and carbon starvation processes co-occurring in dying trees of both species, with the time spent with severe hydraulic failure and carbon starvation, rather than absolute thresholds per se, being a better predictor of impending mortality. Model and empirical data suggest that limited carbon and water exchanges at stomatal, phloem, and below-ground interfaces were associated with mortality of both species. The model-data comparison suggests that the introduction of a mechanistic process into physiology-based models provides equal or improved predictive power over traditional process-model or empirical thresholds. Both biophysical and empirical modeling approaches are useful in understanding processes, particularly when the models fail, because they reveal mechanisms that are likely to underlie mortality. We suggest that for some ecosystems, integration of mechanistic pathogen models into current vegetation models, and evaluation against observations, could result in a breakthrough capability to simulate vegetation dynamics.

Reduced transpiration response to precipitation pulses precedes mortality in a piñon-juniper woodland subject to prolonged drought.

Global climate change is predicted to alter the intensity and duration of droughts, but the effects of changing precipitation patterns on vegetation mortality are difficult to predict. Our objective was to determine whether prolonged drought or above-average precipitation altered the capacity to respond to the individual precipitation pulses that drive productivity and survival. We analyzed 5 yr of data from a rainfall manipulation experiment in piñon-juniper (Pinus edulis-Juniperus monosperma) woodland using mixed effects models of transpiration response to event size, antecedent soil moisture, and post-event vapor pressure deficit. Replicated treatments included irrigation, drought, ambient control and infrastructure control. Mortality was highest under drought, and the reduced post-pulse transpiration in the droughted trees that died was attributable to treatment effects beyond drier antecedent conditions and reduced event size. In particular, trees that died were nearly unresponsive to antecedent shallow soil moisture, suggesting reduced shallow absorbing root area. Irrigated trees showed an enhanced response to precipitation pulses. Prolonged drought initiates a downward spiral whereby trees are increasingly unable to utilize pulsed soil moisture. Thus, the additive effects of future, more frequent droughts may increase drought-related mortality.

Regulation and acclimation of leaf gas exchange in a piñon-juniper woodland exposed to three different precipitation regimes.

Leaf gas-exchange regulation plays a central role in the ability of trees to survive drought, but forecasting the future response of gas exchange to prolonged drought is hampered by our lack of knowledge regarding potential acclimation. To investigate whether leaf gas-exchange rates and sensitivity to drought acclimate to precipitation regimes, we measured the seasonal variations of leaf gas exchange in a mature piñon-juniper Pinus edulis-Juniperus monosperma woodland after 3 years of precipitation manipulation. We compared trees receiving ambient precipitation with those in an irrigated treatment (+30% of ambient precipitation) and a partial rainfall exclusion (-45%). Treatments significantly affected leaf water potential, stomatal conductance and photosynthesis for both isohydric piñon and anisohydric juniper. Leaf gas exchange acclimated to the precipitation regimes in both species. Maximum gas-exchange rates under well-watered conditions, leaf-specific hydraulic conductance and leaf water potential at zero photosynthetic assimilation all decreased with decreasing precipitation. Despite their distinct drought resistance and stomatal regulation strategies, both species experienced hydraulic limitation on leaf gas exchange when precipitation decreased, leading to an intraspecific trade-off between maximum photosynthetic assimilation and resistance of photosynthesis to drought. This response will be most detrimental to the carbon balance of piñon under predicted increases in aridity in the southwestern USA.

Drought predisposes piñon-juniper woodlands to insect attacks and mortality.

To test the hypothesis that drought predisposes trees to insect attacks, we quantified the effects of water availability on insect attacks, tree resistance mechanisms, and mortality of mature piñon pine (Pinus edulis) and one-seed juniper (Juniperus monosperma) using an experimental drought study in New Mexico, USA. The study had four replicated treatments (40 × 40 m plot/replicate): removal of 45% of ambient annual precipitation (H2 O-); irrigation to produce 125% of ambient annual precipitation (H2 O+); a drought control (C) to quantify the impact of the drought infrastructure; and ambient precipitation (A). Piñon began dying 1 yr after drought initiation, with higher mortality in the H2 O- treatment relative to other treatments. Beetles (bark/twig) were present in 92% of dead trees. Resin duct density and area were more strongly affected by treatments and more strongly associated with piñon mortality than direct measurements of resin flow. For juniper, treatments had no effect on insect resistance or attacks, but needle browning was highest in the H2 O- treatment. Our results provide strong evidence that ≥ 1 yr of severe drought predisposes piñon to insect attacks and increases mortality, whereas 3 yr of the same drought causes partial canopy loss in juniper.

Hydraulic limits preceding mortality in a piñon-juniper woodland under experimental drought.

Drought-related tree mortality occurs globally and may increase in the future, but we lack sufficient mechanistic understanding to accurately predict it. Here we present the first field assessment of the physiological mechanisms leading to mortality in an ecosystem-scale rainfall manipulation of a piñon-juniper (Pinus edulis-Juniperus monosperma) woodland. We measured transpiration (E) and modelled the transpiration rate initiating hydraulic failure (E(crit) ). We predicted that isohydric piñon would experience mortality after prolonged periods of severely limited gas exchange as required to avoid hydraulic failure; anisohydric juniper would also avoid hydraulic failure, but sustain gas exchange due to its greater cavitation resistance. After 1 year of treatment, 67% of droughted mature piñon died with concomitant infestation by bark beetles (Ips confusus) and bluestain fungus (Ophiostoma spp.); no mortality occurred in juniper or in control piñon. As predicted, both species avoided hydraulic failure, but safety margins from E(crit) were much smaller in piñon, especially droughted piñon, which also experienced chronically low hydraulic conductance. The defining characteristic of trees that died was a 7 month period of near-zero gas exchange, versus 2 months for surviving piñon. Hydraulic limits to gas exchange, not hydraulic failure per se, promoted drought-related mortality in piñon pine.

Storage and transpiration have negligible effects on delta13C of stem CO2 efflux in large conifer trees.

Stem respiration rates are often quantified by measuring the CO(2) efflux from stems into chambers. It has been suggested that these measurements underestimate respiration because some of the respired CO(2) can be either retained or transported upwards in the transpiration stream. If the stem CO(2) efflux does not represent all respired CO(2), then the interpretation of its isotopic signal may be compromised as well. The C-isotope composition of the respired CO(2) and the measured efflux could differ due to (i) the release of CO(2) produced elsewhere into the stem and transported upwards in xylem water (soil CO(2) or root respired CO(2)); (ii) the retention or release of CO(2) storage pools within the tree stem and (iii) the removal of CO(2) by the transpiration stream. We investigated the effects of these processes in large conifer trees using two manipulative experiments: a labelling experiment and a crown removal experiment. The labelling experiment used an extreme enrichment of dissolved CO(2) in soil water to assess the C uptake by the roots. In this experiment, we found no contamination of the stem CO(2) pool despite clear evidence that the water itself had been taken up. The crown removal experiment tested for vertical CO(2) flux in xylem water by eliminating transpiration. Here, we found no change in the delta(13)C of stem CO(2) efflux (delta(EA); P > 0.05). We concluded that for these large conifers, sap-flow influenced neither delta(13)C of stem efflux nor that of the stem CO(2) pool. By parameterizing Henry's Law for conditions inside the stem, we estimated the transport flux to represent 1-3% of the stem CO(2) efflux to the atmosphere. Finally, assuming a 2 per thousand difference between delta(13)C of root and stem respiration, we estimated that potential contamination of delta(EA) by root respired CO(2) would be < 0.1 per thousand. Thus, neither the release of soil or root CO(2), nor storage in the stem, nor vertical transport of CO(2) in the xylem sap had any detectable influence on delta(13)C of the CO(2) measured in stem efflux.

Nocturnal transpiration causing disequilibrium between soil and stem predawn water potential in mixed conifer forests of Idaho.

Soil water potential (Psi(s)) is often estimated by measuring leaf water potential before dawn (Psi(pd)), based on the assumption that the plant water status has come into equilibrium with that of the soil. However, it has been documented for a number of plant species that stomata do not close completely at night, allowing for nocturnal transpiration and thus preventing nocturnal soil-plant water potential equilibration. The potential for nighttime transpiration necessitates testing the assumption of nocturnal equilibration before accepting Psi(pd) as a valid estimate of Psi(s). We determined the magnitude of disequilibrium between Psi(pd) and Psi(s) in four temperate conifer species across three height classes through a replicated study in northern Idaho. Based on both stomatal conductance and sap flux measurements, we confirmed that the combination of open stomata and high nocturnal atmospheric vapor pressure deficit (D) resulted in nocturnal transpiration in all four species. Nocturnal stomatal conductance (g(s-noc)) averaged about 33% of mid-morning conductance values. We used species-specific estimates of g(s-noc) and leaf specific conductance to correct Psi(pd) values for nocturnal transpiration at the time the samples were collected. Compared with the unadjusted values, corrected values reflected a significantly higher Psi(pd) (when D > 0.12 kPa). These results demonstrate that comparisons of Psi(pd) among species, canopy height classes and sites, and across growing seasons can be influenced by differential amounts of nocturnal transpiration, leading to flawed results. Consequently, it is important to account for the presence of nocturnal transpiration, either through a properly parameterized model or by making Psi(pd) measurements when D is sufficiently low that it cannot drive nocturnal transpiration. Violating these conditions will likely result in underestimation of Psi(s).

Cold air drainage and modeled nocturnal leaf water potential in complex forested terrain.

Spatial variation in microclimate caused by air temperature inversions plays an important role in determining the timing and rate of many physical and biophysical processes. Such phenomena are of particular interest in mountainous regions where complex physiographic terrain can greatly complicate these processes. Recent work has demonstrated that, in some plants, stomata do not close completely at night, resulting in nocturnal transpiration. The following work was undertaken to develop a better understanding of nocturnal cold air drainage and its subsequent impact on the reliability of predawn leaf water potential (Psi(pd)) as a surrogate for soil water potential (Psi(s)). Eight temperature data loggers were installed on a transect spanning a vertical distance of 155 m along a north facing slope in the Mica Creek Experimental Watershed (MCEW) in northern Idaho during July and August 2004. Results indicated strong nocturnal temperature inversions occurring from the low- to upper-mid-slope, typically spanning the lower 88 m of the vertical distance. Based on mean temperatures for both months, inversions resulted in lapse rates of 29.0, 27.0 and 25.0 degrees C km(-1) at 0000, 0400 and 2000 h, respectively. At this scale (i.e., < 1 km), the observed lapse rates resulted in highly variable nighttime vapor pressure deficits (D) over the length of the slope, with variable impacts on modeled disequilibrium between soil and leaf water potential. As a result of cold air drainage, modeled Psi(pd) became consistently more negative (up to -0.3 MPa) at higher elevations during the night based on mean temperatures. Nocturnal inversions on the lower- and mid-slopes resulted in leaf water potentials that were at least 30 and 50% more negative over the lower 88 m of the inversion layer, based on mean and maximum temperatures, respectively. However, on a cloudy night, with low D, the maximum decrease in Psi(pd) was -0.04 MPa. Our results indicate that, given persistent cold air drainage and nighttime stomatal opening, serious errors will result if Psi(s) is estimated from Psi(pd).