Death from hunger or thirst? Phloem death, rather than xylem hydraulic failure, as a driver of fire-induced conifer mortality.

Disruption of photosynthesis and carbon transport due to damage to the tree crown and stem cambial cells, respectively, can cause tree mortality. It has recently been proposed that fire-induced dysfunction of xylem plays an important role in tree mortality. Here, we simultaneously tested the impact of a lethal fire dose on nonstructural carbohydrates (NSCs) and xylem hydraulics in Pinus ponderosa saplings. Saplings were burned with a known lethal fire dose. Nonstructural carbohydrates were assessed in needles, main stems, roots and whole plants, and xylem hydraulic conductivity was measured in the main stems up to 29 d postfire. Photosynthesis and whole plant NSCs declined postfire. Additionally, all burned saplings showed 100% phloem/cambium necrosis, and roots of burned saplings had reduced NSCs compared to unburned and defoliated saplings. We further show that, contrary to patterns observed with NSCs, water transport was unchanged by fire and there was no evidence of xylem deformation in saplings that experienced a lethal dose of heat from fire. We conclude that phloem and cambium mortality, and not hydraulic failure, were probably the causes of death in these saplings. These findings advance our understanding of the physiological response to fire-induced injuries in conifer trees.

Carbon starvation following a decade of experimental drought consumes old reserves in Pinus edulis.

Shifts in the age or turnover time of non-structural carbohydrates (NSC) may underlie changes in tree growth under long-term increases in drought stress associated with climate change. But NSC responses to drought are challenging to quantify, due in part to large NSC stores in trees and subsequently long response times of NSC to climate variation. We measured NSC age (Δ14 C) along with a suite of ecophysiological metrics in Pinus edulis trees experiencing either extreme short-term drought (-90% ambient precipitation plot, 2020-2021) or a decade of severe drought (-45% plot, 2010-2021). We tested the hypothesis that carbon starvation - consumption exceeding synthesis and storage - increases the age of sapwood NSC. One year of extreme drought had no impact on NSC pool size or age, despite significant reductions in predawn water potential, photosynthetic rates/capacity, and twig and needle growth. By contrast, long-term drought halved the age of the sapwood NSC pool, coupled with reductions in sapwood starch concentrations (-75%), basal area increment (-39%), and bole respiration rates (-28%). Our results suggest carbon starvation takes time, as tree carbon reserves appear resilient to extreme disturbance in the short term. However, after a decade of drought, trees apparently consumed old stored NSC to support metabolism.

The osmotic balancing act: When sugars matter for more than metabolism in woody plants.

Sugars and other non-structural carbohydrates are known to serve as currency in plants, to either fuel metabolic activities or as storage for later use. They can also serve non-metabolic purposes of osmoregulation and cryoprotection, especially in perennial woody plants.

Seawater exposure causes hydraulic damage in dying Sitka-spruce trees.

Sea-level rise is one of the most critical challenges facing coastal ecosystems under climate change. Observations of elevated tree mortality in global coastal forests are increasing, but important knowledge gaps persist concerning the mechanism of salinity stress-induced nonhalophytic tree mortality. We monitored progressive mortality and associated gas exchange and hydraulic shifts in Sitka-spruce (Picea sitchensis) trees located within a salinity gradient under an ecosystem-scale change of seawater exposure in Washington State, USA. Percentage of live foliated crown (PLFC) decreased and tree mortality increased with increasing soil salinity during the study period. A strong reduction in gas exchange and xylem hydraulic conductivity (Ks) occurred during tree death, with an increase in the percentage loss of conductivity (PLC) and turgor loss point (πtlp). Hydraulic and osmotic shifts reflected that hydraulic function declined from seawater exposure, and dying trees were unable to support osmotic adjustment. Constrained gas exchange was strongly related to hydraulic damage at both stem and leaf levels. Significant correlations between foliar sodium (Na+) concentration and gas exchange and key hydraulic parameters (Ks, PLC, and πtlp) suggest that cellular injury related to the toxic effects of ion accumulation impacted the physiology of these dying trees. This study provides evidence of toxic effects on the cellular function that manifests in all aspects of plant functioning, leading to unfavourable osmotic and hydraulic conditions.

Short- and long-term effects of fire on stem hydraulics in Pinus ponderosa saplings.

Understanding tree physiological responses to fire is needed to accurately model post-fire carbon processes and inform management decisions. Given trees can die immediately or at extended time periods after fire, we combined two experiments to assess the short- (one-day) and long-term (21-months) fire effects on Pinus ponderosa sapling water transport. Native percentage loss of conductivity (nPLC), vulnerability to cavitation and xylem anatomy were assessed in unburned and burned saplings at lethal and non-lethal fire intensities. Fire did not cause any impact on nPLC and xylem cell wall structure in either experiment. However, surviving saplings evaluated 21-months post-fire were more vulnerable to cavitation. Our anatomical analysis in the long-term experiment showed that new xylem growth adjacent to fire scars had irregular-shaped tracheids and many parenchyma cells. Given conduit cell wall deformation was not observed in the long-term experiment, we suggest that the irregularity of newly grown xylem cells nearby fire wounds may be responsible for decreasing resistance to embolism in burned plants. Our findings suggest that hydraulic failure is not the main short-term physiological driver of mortality for Pinus ponderosa saplings. However, the decrease in embolism resistance in fire-wounded saplings could contribute to sapling mortality in the years following fire.

Foliar respiration is related to photosynthetic, growth and carbohydrate response to experimental drought and elevated temperature.

Short-term plant respiration (R) increases exponentially with rising temperature, but drought could reduce respiration by reducing growth and metabolism. Acclimation may alter these responses. We examined if species with different drought responses would differ in foliar R response to +4.8°C temperature and -45% precipitation in a field experiment with mature piñon and juniper trees, and if any differences between species were related to differences in photosynthesis rates, shoot growth and nonstructural carbohydrates (NSCs). Short-term foliar R had a Q10 of 1.6 for piñon and 2.6 for juniper. Piñon foliar R did not respond to the +4.8°C temperatures, but R increased 1.4× for juniper. Across treatments, piñon foliage had higher growth, lower NSC content, 29% lower photosynthesis rates, and 44% lower R than juniper. Removing 45% precipitation had little impact on R for either species. Species differences in the response of R under elevated temperature were related to substrate availability and stomatal response to leaf water potential. Despite not acclimating to the higher temperature and having higher R than piñon, greater substrate availability in juniper suggests it could supply respiratory demand for much longer than piñon. Species responses will be critical in ecosystem response to a warmer climate.

Hotter droughts alter resource allocation to chemical defenses in piñon pine.

Heat and drought affect plant chemical defenses and thereby plant susceptibility to pests and pathogens. Monoterpenes are of particular importance for conifers as they play critical roles in defense against bark beetles. To date, work seeking to understand the impacts of heat and drought on monoterpenes has primarily focused on young potted seedlings, leaving it unclear how older age classes that are more vulnerable to bark beetles might respond to stress. Furthermore, we lack a clear picture of what carbon resources might be prioritized to support monoterpene synthesis under drought stress. To address this, we measured needle and woody tissue monoterpene concentrations and physiological variables simultaneously from mature piñon pines (Pinus edulis) from a unique temperature and drought manipulation field experiment. While heat had no effect on total monoterpene concentrations, trees under combined heat and drought stress exhibited ~ 85% and 35% increases in needle and woody tissue, respectively, over multiple years. Plant physiological variables like maximum photosynthesis each explained less than 10% of the variation in total monoterpenes for both tissue types while starch and glucose + fructose measured 1-month prior explained ~ 45% and 60% of the variation in woody tissue total monoterpene concentrations. Although total monoterpenes increased under combined stress, some key monoterpenes with known roles in bark beetle ecology decreased. These shifts may make trees more favorable for bark beetle attack rather than well defended, which one might conclude if only considering total monoterpene concentrations. Our results point to cumulative and synergistic effects of heat and drought that may reprioritize carbon allocation of specific non-structural carbohydrates toward defense.

Conifers depend on established roots during drought: results from a coupled model of carbon allocation and hydraulics.

Trees may survive prolonged droughts by shifting water uptake to reliable water sources, but it is unknown if the dominant mechanism involves activating existing roots or growing new roots during drought, or some combination of the two. To gain mechanistic insights on this unknown, a dynamic root-hydraulic modeling framework was developed that set up a feedback between hydraulic controls over carbon allocation and the role of root growth on soil-plant hydraulics. The new model was tested using a 5 yr drought/heat field experiment on an established piñon-juniper stand with root access to bedrock groundwater. Owing to the high carbon cost per unit root area, modeled trees initialized without adequate bedrock groundwater access experienced potentially lethal declines in water potential, while all of the experimental trees maintained nonlethal water potentials. Simulated trees were unable to grow roots rapidly enough to mediate the hydraulic stress, particularly during warm droughts. Alternatively, modeled trees initiated with root access to bedrock groundwater matched the hydraulics of the experimental trees by increasing their water uptake from bedrock groundwater when soil layers dried out. Therefore, the modeling framework identified a critical mechanism for drought response that required trees to shift water uptake among existing roots rather than growing new roots.

Tree defence and bark beetles in a drying world: carbon partitioning, functioning and modelling.

Drought has promoted large-scale, insect-induced tree mortality in recent years, with severe consequences for ecosystem function, atmospheric processes, sustainable resources and global biogeochemical cycles. However, the physiological linkages among drought, tree defences, and insect outbreaks are still uncertain, hindering our ability to accurately predict tree mortality under on-going climate change. Here we propose an interdisciplinary research agenda for addressing these crucial knowledge gaps. Our framework includes field manipulations, laboratory experiments, and modelling of insect and vegetation dynamics, and focuses on how drought affects interactions between conifer trees and bark beetles. We build upon existing theory and examine several key assumptions: (1) there is a trade-off in tree carbon investment between primary and secondary metabolites (e.g. growth vs defence); (2) secondary metabolites are one of the main component of tree defence against bark beetles and associated microbes; and (3) implementing conifer-bark beetle interactions in current models improves predictions of forest disturbance in a changing climate. Our framework provides guidance for addressing a major shortcoming in current implementations of large-scale vegetation models, the under-representation of insect-induced tree mortality.

Dead or dying? Quantifying the point of no return from hydraulic failure in drought-induced tree mortality.

Determining physiological mechanisms and thresholds for climate-driven tree die-off could help improve global predictions of future terrestrial carbon sinks. We directly tested for the lethal threshold in hydraulic failure - an inability to move water due to drought-induced xylem embolism - in a pine sapling experiment. In a glasshouse experiment, we exposed loblolly pine (Pinus taeda) saplings (n = 83) to drought-induced water stress ranging from mild to lethal. Before rewatering to relieve drought stress, we measured native hydraulic conductivity and foliar color change. We monitored all measured individuals for survival or mortality. We found a lethal threshold at 80% loss of hydraulic conductivity - a point of hydraulic failure beyond which it is more likely trees will die, than survive, and describe mortality risk across all levels of water stress. Foliar color changes lagged behind hydraulic failure - best predicting when trees had been dead for some time, rather than when they were dying. Our direct measurement of native conductivity, while monitoring the same individuals for survival or mortality, quantifies a continuous probability of mortality risk from hydraulic failure. Predicting tree die-off events and understanding the mechanism involved requires knowledge not only of when trees are dead, but when they begin dying - having passed the point of no return.

Research frontiers for improving our understanding of drought-induced tree and forest mortality.

Accumulating evidence highlights increased mortality risks for trees during severe drought, particularly under warmer temperatures and increasing vapour pressure deficit (VPD). Resulting forest die-off events have severe consequences for ecosystem services, biophysical and biogeochemical land-atmosphere processes. Despite advances in monitoring, modelling and experimental studies of the causes and consequences of tree death from individual tree to ecosystem and global scale, a general mechanistic understanding and realistic predictions of drought mortality under future climate conditions are still lacking. We update a global tree mortality map and present a roadmap to a more holistic understanding of forest mortality across scales. We highlight priority research frontiers that promote: (1) new avenues for research on key tree ecophysiological responses to drought; (2) scaling from the tree/plot level to the ecosystem and region; (3) improvements of mortality risk predictions based on both empirical and mechanistic insights; and (4) a global monitoring network of forest mortality. In light of recent and anticipated large forest die-off events such a research agenda is timely and needed to achieve scientific understanding for realistic predictions of drought-induced tree mortality. The implementation of a sustainable network will require support by stakeholders and political authorities at the international level.

Stem radial growth and water storage responses to heat and drought vary between conifers with differing hydraulic strategies.

We investigated stem radial growth and water storage dynamics of 2 conifer species differing in hydraulic carbon strategies, Juniperus monosperma and Pinus edulis, under conditions of ambient, drought (∼45% reduction in precipitation), heat (∼4.8 °C temperature increase), and the combination of drought + heat, in 2013 and 2014. Juniper maintained low growth across all treatments. Overall, the relatively isohydric piñon pine showed significantly greater growth and water storage recharge than the relatively anisohydric juniper across all treatments in the average climate year (2014) but no differences in the regionally dry year (2013). Piñon pine ceased growth at a constant predawn water potential across all treatments and at a less negative water potential threshold than juniper. Heat has a greater negative impact on piñon pines' growth and water storage than drought, whereas juniper was, in contrast, unaffected by heat but strongly impacted by drought. The whole-plant hydraulic carbon strategies, in this case captured using the isohydric/anisohydric concept, translate into alternative growth and water storage strategies under drought and heat conditions.

Belowground annual ring growth coordinates with aboveground phenology and timing of carbon storage in two tallgrass prairie forb species.

PREMISE OF THE STUDY: Herb chronology, the study of belowground annual growth rings in perennial forbs, has much potential as a tool for monitoring plant growth as a function of environment. To harness this potential, understanding of the coordination between ring ontogeny, aboveground phenology, and the temporal allocation of carbon products belowground in herbaceous forbs must be improved. METHODS: We investigated these relationships in two southern United States tallgrass prairie perennial forb species, Asclepias viridis and Lespedeza stuevei, making monthly excavations for a year. KEY RESULTS: Belowground xylogenesis began when starch reserves were at their seasonal low in the spring as shoots reached maximum height. The highest relative radial growth of the ring occurred concurrently with replenishment of root starch reserves in early summer. Xylogenesis concluded with leaf senescence in late summer and belowground starch reserves near saturation. CONCLUSIONS: By demonstrating that ring ontogeny is tied to early summer starch replenishment, our results illustrate the mechanisms behind previous findings where ring width was highly correlated with summer climatic conditions for these two species. This study provides a new physiological link between how ring chronologies in herbs often accord with growing-season environment; further dissecting this phenomenon is vital in unlocking the potential of herb chronology.

Warming combined with more extreme precipitation regimes modifies the water sources used by trees.

The persistence of vegetation under climate change will depend on a plant's capacity to exploit water resources. We analyzed water source dynamics in piñon pine and juniper trees subjected to precipitation reduction, atmospheric warming, and to both simultaneously. Piñon and juniper exhibited different and opposite shifts in water uptake depth in response to experimental stress and background climate over 3 yr. During a dry summer, juniper responded to warming with a shift to shallow water sources, whereas piñon pine responded to precipitation reduction with a shift to deeper sources in autumn. In normal and wet summers, both species responded to precipitation reduction, but juniper increased deep water uptake and piñon increased shallow water uptake. Shifts in the utilization of water sources were associated with reduced stomatal conductance and photosynthesis, suggesting that belowground compensation in response to warming and water reduction did not alleviate stress impacts for gas exchange. We have demonstrated that predicted climate change could modify water sources of trees. Warming impairs juniper uptake of deep sources during extended dry periods. Precipitation reduction alters the uptake of shallow sources following extended droughts for piñon. Shifts in water sources may not compensate for climate change impacts on tree physiology.

Responses of two semiarid conifer tree species to reduced precipitation and warming reveal new perspectives for stomatal regulation.

Relatively anisohydric species are predicted to be more predisposed to hydraulic failure than relatively isohydric species, as they operate with narrower hydraulic safety margins. We subjected co-occurring anisohydric Juniperus monosperma and isohydric Pinus edulis trees to warming, reduced precipitation, or both, and measured their gas exchange and hydraulic responses. We found that reductions in stomatal conductance and assimilation by heat and drought were more frequent during relatively moist periods, but these effects were not exacerbated in the combined heat and drought treatment. Counter to expectations, both species exhibited similar gs temporal dynamics in response to drought. Further, whereas P. edulis exhibited chronic embolism, J. monosperma showed very little embolism due to its conservative stomatal regulation and maintenance of xylem water potential above the embolism entry point. This tight stomatal control and low levels of embolism experienced by juniper refuted the notion that very low water potentials during drought are associated with loose stomatal control and with the hypothesis that anisohydric species are more prone to hydraulic failure than isohydric species. Because direct association of stomatal behaviour with embolism resistance can be misleading, we advocate consideration of stomatal behaviour relative to embolism resistance for classifying species drought response strategies.

Drought and resprouting plants.

Many species have the ability to resprout vegetatively after a substantial loss of biomass induced by environmental stress, including drought. Many of the regions characterised by ecosystems where resprouting is common are projected to experience more frequent and intense drought during the 21st Century. However, in assessments of ecosystem response to drought disturbance there has been scant consideration of the resilience and post-drought recovery of resprouting species. Systematic differences in hydraulic and allocation traits suggest that resprouting species are more resilient to drought-stress than nonresprouting species. Evidence suggests that ecosystems dominated by resprouters recover from disturbance more quickly than ecosystems dominated by nonresprouters. The ability of resprouters to avoid mortality and withstand drought, coupled with their ability to recover rapidly, suggests that the impact of increased drought stress in ecosystems dominated by these species may be small. The strategy of resprouting needs to be modelled explicitly to improve estimates of future climate-change impacts on the carbon cycle, but this will require several important knowledge gaps to be filled before resprouting can be properly implemented.

Experimental drought and heat can delay phenological development and reduce foliar and shoot growth in semiarid trees.

Higher temperatures associated with climate change are anticipated to trigger an earlier start to the growing season, which could increase the terrestrial C sink strength. Greater variability in the amount and timing of precipitation is also expected with higher temperatures, bringing increased drought stress to many ecosystems. We experimentally assessed the effects of higher temperature and drought on the foliar phenology and shoot growth of mature trees of two semiarid conifer species. We exposed field-grown trees to a ~45% reduction in precipitation with a rain-out structure ('drought'), a ~4.8 °C temperature increase with open-top chambers ('heat'), and a combination of both simultaneously ('drought + heat'). Over the 2013 growing season, drought, heat, and drought + heat treatments reduced shoot and needle growth in piñon pine (Pinus edulis) by ≥39%, while juniper (Juniperus monosperma) had low growth and little response to these treatments. Needle emergence on primary axis branches of piñon pine was delayed in heat, drought, and drought + heat treatments by 19-57 days, while secondary axis branches were less likely to produce needles in the heat treatment, and produced no needles at all in the drought + heat treatment. Growth of shoots and needles, and the timing of needle emergence correlated inversely with xylem water tension and positively with nonstructural carbohydrate concentrations. Our findings demonstrate the potential for delayed phenological development and reduced growth with higher temperatures and drought in tree species that are vulnerable to drought and reveal potential mechanistic links to physiological stress responses. Climate change projections of an earlier and longer growing season with higher temperatures, and consequent increases in terrestrial C sink strength, may be incorrect for regions where plants will face increased drought stress with climate change.

Nonstructural leaf carbohydrate dynamics of Pinus edulis during drought-induced tree mortality reveal role for carbon metabolism in mortality mechanism.

Vegetation change is expected with global climate change, potentially altering ecosystem function and climate feedbacks. However, causes of plant mortality, which are central to vegetation change, are understudied, and physiological mechanisms remain unclear, particularly the roles of carbon metabolism and xylem function. We report analysis of foliar nonstructural carbohydrates (NSCs) and associated physiology from a previous experiment where earlier drought-induced mortality of Pinus edulis at elevated temperatures was associated with greater cumulative respiration. Here, we predicted faster NSC decline for warmed trees than for ambient-temperature trees. Foliar NSC in droughted trees declined by 30% through mortality and was lower than in watered controls. NSC decline resulted primarily from decreased sugar concentrations. Starch initially declined, and then increased above pre-drought concentrations before mortality. Although temperature did not affect NSC and sugar, starch concentrations ceased declining and increased earlier with higher temperatures. Reduced foliar NSC during lethal drought indicates a carbon metabolism role in mortality mechanism. Although carbohydrates were not completely exhausted at mortality, temperature differences in starch accumulation timing suggest that carbon metabolism changes are associated with time to death. Drought mortality appears to be related to temperature-dependent carbon dynamics concurrent with increasing hydraulic stress in P. edulis and potentially other similar species.

No carbon storage in growth-limited trees in a semi-arid woodland.

Plant survival depends on a balance between carbon supply and demand. When carbon supply becomes limited, plants buffer demand by using stored carbohydrates (sugar and starch). During drought, NSCs (non-structural carbohydrates) may accumulate if growth stops before photosynthesis. This expectation is pervasive, yet few studies have combined simultaneous measurements of drought, photosynthesis, growth, and carbon storage to test this. Using a field experiment with mature trees in a semi-arid woodland, we show that growth and photosynthesis slow in parallel as [Formula: see text] declines, preventing carbon storage in two species of conifer (J. monosperma and P. edulis). During experimental drought, growth and photosynthesis were frequently co-limited. Our results point to an alternative perspective on how plants use carbon that views growth and photosynthesis as independent processes both regulated by water availability.

Fire intensity impacts on physiological performance and mortality in Pinus monticola and Pseudotsuga menziesii saplings: a dose-response analysis.

Fire is a major cause of tree injury and mortality worldwide, yet our current understanding of fire effects is largely based on ocular estimates of stem charring and foliage discoloration, which are error prone and provide little information on underlying tree function. Accurate quantification of physiological performance is a research and forest management need, given that declining performance could help identify mechanisms of-and serve as an early warning sign for-mortality. Many previous efforts have been hampered by the inability to quantify the heat flux that a tree experiences during a fire, given its highly variable nature in space and time. In this study, we used a dose-response approach to elucidate fire impacts by subjecting Pinus monticola var. minima Lemmon and Pseudotsuga menziesii (Mirb.) Franco var. glauca (Beissn.) Franco saplings to surface fires of varying intensity doses and measuring short-term post-fire physiological performance in photosynthetic rate and chlorophyll fluorescence. We also evaluated the ability of spectral reflectance indices to quantify change in physiological performance at the individual tree crown and stand scales. Although physiological performance in both P. monticola and P. menziesii declined with increasing fire intensity, P. monticola maintained a greater photosynthetic rate and higher chlorophyll fluorescence at higher doses, for longer after the fire. Pinus monticola also had complete survival at lower fire intensity doses, whereas P. menziesii had some mortality at all doses, implying higher fire resistance for P. monticola at this life stage. Generally, individual-scale spectral indices were more accurate at quantifying physiological performance than those acquired at the stand-scale. The Photochemical Reflectance Index outperformed other indices at quantifying photosynthesis and chlorophyll fluorescence, highlighting its potential use to quantify crown scale physiological performance. Spectral indices that incorporated near-infrared and shortwave infrared reflectance, such as the Normalized Burn Ratio, were accurate at characterizing stand-scale mortality. The results from this study were included in a conifer cross-comparison using physiology and mortality data from other dose-response studies. The comparison highlights the close evolutionary relationship between fire and species within the Pinus genus, assessed to date, given the high survivorship of Pinus species at lower fire intensities versus other conifers.