Temperature dependence of pollen germination and tube growth in conifers relates to their distribution along an elevational gradient in Washington State, USA.

BACKGROUND AND AIMS: Pollen germination and tube growth are essential processes for successful fertilization. They are among the most temperature-vulnerable stages and subsequently affect seed production and determine population persistence and species distribution under climate change. Our study aims to investigate intra- and inter-specific variations in the temperature dependence of pollen germination and tube length growth and to explore how these variations differ for pollen from elevational gradients. METHODS: We focused on three conifer species, Pinus contorta, Picea engelmannii, and Pinus ponderosa, with pollen collected from 350 to 2200m elevation in Washington State, USA. We conducted pollen viability tests at temperatures from 5 to 40°C in 5°C intervals. After testing for four days, we took images of these samples under a microscope to monitor pollen germination percentage (GP) and tube length (TL). We applied the Gamma function to describe the temperature dependence of GP and TL and estimated key parameters, including the optimal temperature for GP (Topt_GP) and TL (Topt_TL). KEY RESULTS: Results showed that pollen from three species and different elevations within a species have different GP, TL, Topt_GP, and Topt_TL. The population with a higher Topt_GP would also have a higher Topt_TL, while Topt_TL was generally higher than Topt_GP, i.e., a positive but not one-to-one relationship. However, only Pinus contorta showed that populations from higher elevations have lower Topt_GP and Topt_TL and vice versa. The variability in GP increased at extreme temperatures, whereas the variability in TL was greatest near Topt_TL. CONCLUSIONS: Our study demonstrates the temperature dependences of three conifers across a wide range of temperatures. Pollen germination and tube growth are highly sensitive to temperature conditions and vary among species and elevations, affecting their reproduction success during warming. Our findings can provide valuable insights to advance our understanding of how conifer pollen responds to rising temperatures.

Long-term frequent fire and cattle grazing alter dry forest understory vegetation.

Understanding fire and large herbivore interactions in interior western forests is critical, owing to the extensive and widespread co-occurrence of these two disturbance types and multiple present and future implications for forest resilience, conservation and restoration. However, manipulative studies focused on interactions and outcomes associated with these two disturbances are rare in forested rangelands. We investigated understory vegetation response to 5-year spring and fall prescribed fire and domestic cattle grazing exclusion in ponderosa pine stands and reported long-term responses, almost two decades after the first entry fires. In fall burn areas open to cattle grazing, total understory cover prior to utilization was about 12% lower compared with fall burn areas where cattle were experimentally excluded. This response was not strongly driven by a particular palatable or unpalatable plant functional group. Fire and grazing are likely interacting in a numerically mediated process, as we found little evidence to support a functionally moderated pathway. Post-fire green-up may equalize forage to a certain extent and concentrate herbivores in the smaller burned areas within pastures, constraining a positive understory response to burning. Fall fire and grazing also increased annual forbs and resprouting shrubs. The effects of spring burning were relatively minor, and we found no interaction with grazing. The nonnative annual grass Bromus tectorum (cheatgrass) remains a problematic invader linked to fall burning but not grazing in stands that had higher propagule pressure when the experiment was initiated. At these sites, exotic grass was a major component of the vegetation by 2015, and invasion was also increasing in spring burn and unburned areas. Information from our study suggests that frequent fall fires and cattle grazing combined may reduce understory resilience in similar dry ponderosa pine forests. Consideration of longer fire return intervals, resting areas after fire, virtual fencing, or burning entire pastures may help to mitigate the effects noted in this study.

Long-term efficacy of fuel reduction and restoration treatments in Northern Rockies dry forests.

Fuel and restoration treatments seeking to mitigate the likelihood of uncharacteristic high-severity wildfires in forests with historically frequent, low-severity fire regimes are increasingly common, but long-term treatment effects on fuels, aboveground carbon, plant community structure, ecosystem resilience, and other ecosystem attributes are understudied. We present 20-year responses to thinning and prescribed burning treatments commonly used in dry, low-elevation forests of the western United States from a long-term study site in the Northern Rockies that is part of the National Fire and Fire Surrogate Study. We provide a comprehensive synthesis of short-term (<4 years) and mid-term (<14 years) results from previous findings. We then place these results in the context of a mountain pine beetle (MPB; Dendroctonus ponderosae) outbreak that impacted the site 5-10 years post-treatment and describe 20-year responses to assess the longevity of restoration and fuel reduction treatments in light of the MPB outbreak. Thinning treatments had persistently lower forest density and higher tree growth, but effects were more pronounced when thinning was combined with prescribed fire. The thinning+prescribed fire treatment had the additional benefit of maintaining the highest proportion of ponderosa pine (Pinus ponderosa) for overstory and regeneration. No differences in understory native plant cover and richness or exotic species cover remained after 20 years, but exotic species richness, while low relative to native species, was still higher in the thinning+prescribed fire treatment than the control. Aboveground live carbon stocks in thinning treatments recovered to near control and prescribed fire treatment levels by 20 years. The prescribed fire treatment and control had higher fuel loads than thinning treatments due to interactions with the MPB outbreak. The MPB-induced changes to forest structure and fuels increased the fire hazard 20 years post-treatment in the control and prescribed fire treatment. Should a wildfire occur now, the thinning+prescribed fire treatment would likely have the lowest intensity fire and highest tree survival and stable carbon stocks. Our findings show broad support that thinning and prescribed fire increase ponderosa pine forest resilience to both wildfire and bark beetles for up to 20 years, but efficacy is waning and additional fuel treatments are needed to maintain resilience.

Ability of seedlings to survive heat and drought portends future demographic challenges for five southwestern US conifers.

Climate change and disturbance are altering forests and the rates and locations of tree regeneration. In semi-arid forests of the southwestern USA, limitations imposed by hot and dry conditions are likely to influence seedling survival. We examined how the survival of 1-year seedlings of five southwestern US conifer species whose southwestern distributions range from warmer and drier woodlands and forests (Pinus edulis Engelm., Pinus ponderosa Douglas ex C. Lawson) to cooler and wetter subalpine forests (Pseudotsuga menziesii (Mirb.) Franco, Abies concolor (Gord. & Glend.) Lindl. Ex Hildebr. and Picea engelmannii Parry ex Engelm.) changed in response to low moisture availability, high temperatures and high vapor pressure deficit in incubators. We used a Bayesian framework to construct discrete-time proportional hazard models that explained 55-75% of the species-specific survival variability. We applied these to the recent climate (1980-2019) of the southwestern USA as well as 1980-2099 CMIP5 climate projections with the RCP8.5 emissions pathway. We found that the more mesic species (i.e., P. menziesii, A. concolor and P. engelmannii) were more susceptible to the effects of hot and dry periods. However, their existing ranges are not projected to experience the conditions we tested as early in the 21st century as the more xeric P. edulis and P. ponderosa, leading to lower percentages of their existing ranges predicted to experience seedling-killing conditions. By late-century, extensive areas of each species southwestern range could experience climate conditions that increase the likelihood of seedling mortality. These results demonstrate that empirically derived physiological limitations can be used to inform where species composition or vegetation type change are likely to occur in the southwestern USA.

Nonstructural carbohydrates explain post-fire tree mortality and recovery patterns.

Trees use nonstructural carbohydrates (NSCs) to support many functions, including recovery from disturbances. However, NSC's importance for recovery following fire and whether NSC depletion contributes to post-fire delayed mortality are largely unknown. We investigated how fire affects NSCs based on fire-caused injury from a prescribed fire in a young Pinus ponderosa (Lawson & C. Lawson) stand. We assessed crown injury (needle scorch and bud kill) and measured NSCs of needles and inner bark (i.e., secondary phloem) of branches and main stems of trees subject to fire and at an adjacent unburned site. We measured NSCs pre-fire and at six timesteps post-fire (4 days-16 months). While all trees initially survived the fire, NSC concentrations declined quickly in burned trees relative to unburned controls over the same post-fire period. This decline was strongest for trees that eventually died, but those that survived recovered to unburned levels within 14 months post-fire. Two months post-fire, the relationship between crown scorch and NSCs of the main stem inner bark was strongly negative (Adj-R2 = 0.83). Our results support the importance of NSCs for tree survival and recovery post-fire and suggest that post-fire NSC depletion is in part related to reduced photosynthetic leaf area that subsequently limits carbohydrate availability for maintaining tree function. Crown scorch is a commonly measured metric of tree-level fire severity and is often linked to post-fire tree outcome (i.e., recovery or mortality). Thus, our finding that NSC depletion may be the mechanistic link between the fire-caused injury and tree outcome will help improve models of post-fire tree mortality and forest recovery.

Increased aridity is associated with stronger tradeoffs in ponderosa pine vital functions.

Trees must allocate resources to core functions like growth, defense, and reproduction. These allocation patterns have profound effects on forest health, yet little is known about how core functions trade off over time, and even less is known about how a changing climate will impact tradeoffs. We conducted a 21-year survey of growth, defense, and reproduction in 80 ponderosa pine individuals spanning eight populations across environmental gradients along the Colorado Front Range, USA. We used linear mixed models to describe tradeoffs among these functions and to characterize variability among and within individuals over time. Growth and defense were lower in years of high cone production, and local drought conditions amplified year-to-year tradeoffs between reproduction and growth, where trees located at sites with hotter and drier climates showed stronger tradeoffs between reproduction and growth. Our results support the environmental stress hypothesis of masting, which predicts that greater interannual variation in tree functions will be associated with more marginal environments, such as those that are prone to drought. With warming temperatures and increased exposure to drought stress, trees will be faced with stronger interannual tradeoffs, which could lead to further decreases in growth and defensive efforts, ultimately increasing risks of mortality.

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.

Drivers of population differentiation in phenotypic plasticity in a temperate conifer: A 27-year study.

Phenotypic plasticity is a main mechanism for organisms to cope with changing environments and broaden their ecological range. Plasticity is genetically based and can evolve under natural selection, such that populations within a species show distinct phenotypic responses to the environment if evolved under different conditions. Understanding how intraspecific variation in phenotypic plasticity arises is critical to assess potential adaptation to ongoing climate change. Theory predicts that plasticity is favored in more favorable but variable environments. Yet, many theoretical predictions about benefits, costs, and selection on plasticity remain untested. To test these predictions, we took advantage of three genetic trials in the northern Rocky Mountains, USA, which assessed 23 closely located Pinus ponderosa populations over 27 years. Mean environmental conditions and their spatial patterns of variation at the seed source populations were characterized based on six basic climate parameters. Despite the small area of origin, there was significant genetic variation in phenotypic plasticity for tree growth among populations. We found a significant negative correlation between phenotypic plasticity and the patch size of environmental heterogeneity at the seed source populations, but not with total environmental spatial variance. These results show that populations exposed to high microhabitat heterogeneity have evolved higher phenotypic plasticity and that the trigger was the grain rather than the total magnitude of spatial heterogeneity. Contrary to theoretical predictions, we also found a positive relationship between population plasticity and summer drought at the seed source, indicating that drought can act as a trigger of plasticity. Finally, we found a negative correlation between the quantitative genetic variance within populations and their phenotypic plasticity, suggesting compensatory adaptive mechanisms for the lack of genetic diversity. These results improve our understanding of the microevolutionary drivers of phenotypic plasticity, a critical process for resilience of long-lived species under climate change, and support decision-making in tree genetic improvement programs and seed transfer strategies.

Forest restoration treatments increased growth and did not change survival of ponderosa pines in severe drought, Arizona.

We report on survival and growth of ponderosa pines (Pinus ponderosa Douglas ex P. Lawson & C. Lawson) 2 decades after forest restoration treatments in the G. A. Pearson Natural Area, northern Arizona. Despite protection from harvest that conserved old trees, a dense forest susceptible to uncharacteristically severe disturbance had developed during more than a century of exclusion of the previous frequent surface-fire regime that ceased upon Euro-American settlement in approximately 1876. Trees were thinned in 1993 to emulate prefire-exclusion forest conditions, accumulated forest floor was removed, and surface fire was re-introduced at 4-years intervals (full restoration). There was also a partial restoration treatment consisting of thinning alone. Compared with untreated controls, mortality of old trees (mean age 243 years, maximum 462 years) differed by <1 tree ha-1 and old-tree survival was statistically indistinguishable between treatments (90.5% control, 92.3% full, 82.6% partial). Post-treatment growth as measured by basal area increment of both old (pre-1876) and young (post-1876) pines was significantly higher in both treatments than counterpart control trees for more than 2 decades following thinning. Drought meeting the definition of megadrought affected the region almost all the time since the onset of the experiment, including 3 years that were severely dry. Growth of all trees declined in the driest 3 years, but old and young treated trees had significantly less decline. Association of tree growth with temperature (negative correlation) and precipitation (positive correlation) was much weaker in treated trees, indicating that they may experience less growth decline from warmer, drier conditions predicted in future decades. Overall, tree responses after the first 2 decades following treatment suggest that forest restoration treatments have led to substantial, sustained improvement in the growth of old and young ponderosa pines without affecting old-tree survival, thereby improving resilience to a warming climate.

Tree growth sensitivity to climate varies across a seasonal precipitation gradient.

Spatial patterns of precipitation in the southwestern United States result in a complex gradient from winter-to-summer moisture dominance that influences tree growth. In response, tree growth exhibits seasonal-to-annual variability that is evident in the growth of whole tree rings, and in sub-annual sections such as earlywood and latewood. We evaluated the influence of precipitation and temperature on the growth of Pinus ponderosa trees in 11 sites in the southwestern US. Precipitation during the year of growth and the prior year accounted for about half of the climate influence on annual growth, with the other half reflecting conditions 2-4 years prior to growth, indicating that individual trees do indeed exhibit multi-year "memory" of climate. Trees in wetter sites exhibited weaker influence of past precipitation inputs, but longer memory of climatic variability. Conversely, trees in dry sites exhibited shorter memory of long-term climatic variability, but greater sensitivity to past precipitation effects. These results are consistent with the existence of complex interactions between endogenous (phenotype) effects and exogenous (climate) effects in controlling climate memory in trees. After accounting for climate, residual variability in latewood growth was negatively correlated with earlywood growth, indicating a potential tradeoff between latewood versus earlywood growth. This study provides new insights that will assist the accurate prediction of woody biomass growth and forest carbon sequestration across a southwestern US precipitation gradient.

Low- and moderate-severity fire offers key insights for landscape restoration in ponderosa pine forests.

Restoration goals in fire-prone conifer forests include mitigating fire hazard while restoring forest structural components linked to disturbance resilience and ecological function. Restoration of overstory spatial pattern in forests often falls short of management objectives due to complexities in implementation, regulation, and available data. When historical data is available, it is often collected at plots too small to inform coarse-scale metrics like gap size and structure of tree patches (e.g., 1 ha). Principles of ecological forestry typically emphasize overstory removal patterns that emulate those of natural disturbances. So, low- and moderate-severity portions of contemporary wildfires may serve as a guide to restoration treatments where mixed-severity fires occur. Here, we compare forest spatial pattern and configuration in 15 mechanical restoration treatments and low- and moderate-severity portions of three wildfires in ponderosa pine-dominated forests to determine how they differ in spatial pattern. We obtained satellite imagery of restoration treatments and wildfires and used supervised classification to differentiate canopy and openings. We assessed elements of landscape structure including canopy and gap cover, gap attributes, and landscape heterogeneity for each disturbance type. We found that both mechanical restoration treatments and low- and moderate-severity portions of wildfires reduced forest cover, increased gap cover, and altered pattern and arrangement of gaps relative to undisturbed areas, though the magnitude of changes were greatest in the burned sites. Low- and moderate-severity wildfire consistently increased landscape heterogeneity, but mechanical treatments did not. This suggests that a greater emphasis on increasing gap and patch spatial structure may make mechanical treatments more congruent with natural disturbances. Outcomes of low- and moderate-severity portions of wildfires may provide important information upon which to base management prescriptions where reference data on landscape patterns is unavailable.

Combined drought and bark beetle attacks deplete non-structural carbohydrates and promote death of mature pine trees.

How carbohydrate reserves in conifers respond to drought and bark beetle attacks are poorly understood. We investigated changes in carbohydrate reserves and carbon-dependent diterpene defences in ponderosa pine trees that were experimentally subjected to two levels of drought stress (via root trenching) and two types of biotic challenge treatments (pheromone-induced bark beetle attacks or inoculations with crushed beetles that include beetle-associated fungi) for two consecutive years. Our results showed that trenching did not influence carbohydrates, whereas both biotic challenges reduced amounts of starch and sugars of trees. However, only the combined trenched-bark beetle attacked trees depleted carbohydrates and died during the first year of attacks. While live trees contained higher carbohydrates than dying trees, amounts of constitutive and induced diterpenes produced did not vary between live and beetle-attacked dying trees, respectively. Based on these results we propose that reallocation of carbohydrates to diterpenes during the early stages of beetle attacks is limited in drought-stricken trees, and that the combination of biotic and abiotic stress leads to tree death. The process of tree death is subsequently aggravated by beetle girdling of phloem, occlusion of vascular tissue by bark beetle-vectored fungi, and potential exploitation of host carbohydrates by bark beetle symbionts as nutrients.

The effects of ENSO and the North American monsoon on mast seeding in two Rocky Mountain conifer species.

We aimed to disentangle the patterns of synchronous and variable cone production (i.e. masting) and its relationship to climate in two conifer species native to dry forests of western North America. We used cone abscission scars to reconstruct ca 15 years of recent cone production in Pinus edulis and Pinus ponderosa, and used redundancy analysis to relate time series of annual cone production to climate indices describing the North American monsoon and the El Niño Southern Oscillation (ENSO). We show that the sensitivity to climate and resulting synchrony in cone production varies substantially between species. Cone production among populations of P. edulis was much more spatially synchronous and more closely related to large-scale modes of climate variability than among populations of P. ponderosa. Large-scale synchrony in P. edulis cone production was associated with the North American monsoon and we identified a dipole pattern of regional cone production associated with ENSO phase. In P. ponderosa, these climate indices were not strongly associated with cone production, resulting in asynchronous masting patterns among populations. This study helps frame our understanding of mast seeding as a life-history strategy and has implications for our ability to forecast mast years in these species. This article is part of the theme issue 'The ecology and evolution of synchronized seed production in plants'.

Understanding introduction history: Genetic structure and diversity of the edible ectomycorrhizal fungus, Suillus luteus, in Patagonia (Argentina).

Suillus luteus is a common ectomycorrhizal (EM) fungus associated with several Pinus species. It is distributed throughout the Northern Hemisphere and has been introduced into South America and New Zealand. We examined the genetic structure and population biology of S.luteus, which was introduced into Patagonian plantations with Pinus species in Argentina. Overall, 106 samples were collected at 11 geographically separated sites (i.e., Pinus plantations) along a latitudinal gradient in Patagonia (ca. 38°-46° south latitude). Phylogenetic analyses confirmed placement in S.luteus. Genetic analysis demonstrated moderate within-site genetic diversity, but low differentiation between sites. No clear clusters were detected geographically or in relation to host species of Pinus. Our results suggest that the weak genetic structure of the species reflects the short time that has elapsed since the introduction of S.luteus into Patagonia, and its expansion with exotic afforestation there. Moreover, the lack of structure is consistent with a founder effect, suggesting the introduction of a small number of genets that spread throughout all the plantations. Therefore, the high level of gene flow and weak genetic structure observed are probably related to the anthropogenic movement of inoculum associated with forestry practices.

Bimodal cambial activity and false-ring formation in conifers under a monsoon climate.

Tracking wood formation in semiarid regions during the seasonal march of precipitation extremes has two important applications. It can provide (i) insight into the adaptive capacities of trees to drought and (ii) a basis for a richer interpretation of tree-ring data, assisting in a deeper understanding of past and current climate. In the southwestern USA, the anatomical signature of seasonally bimodal precipitation is the 'false ring'-a band of latewood-like cells in the earlywood. These occur when a particularly deep drought during the early growing season ends abruptly with timely, mid-growing season monsoonal rains. Such conditions presented in southern Arizona in 2014, enabling us to explore false-ring formation in ponderosa pine (Pinus ponderosa Lawson and C. Lawson) and Douglas-fir (Pseudotsuga menziesii Mirb. Franco) in mixed-conifer forest at 2573 m above sea level. We ask: what were the cell-by-cell timings and durations in the phases of wood cell development in 2014? How do these seasonal patterns relate to strongly fluctuating environmental conditions during the growing season? We took weekly microcores from March through November from six ponderosa pine and seven Douglas-fir trees at a well-instrumented flux tower site. Thin sections were prepared, and we counted cells in cambial, expansion, cell wall thickening and mature phases. For ponderosa pine trees forming a false ring, the first impact of intensifying seasonal drought was seen in the enlarging phase and then, almost a month later, in cambial activity. In this species, recovery from drought was associated with recovery first in cambial activity, followed by cell enlargement. This timing raised the possibility that cell division may be affected by atmospheric moisture increases before soil recharge. In both species, the last false-ring cells matured during the summer rainy season. Bimodal cambial activity coincident with moisture availability was observed in both species, whether or not they formed a false ring. This deeper knowledge of the precise timing of both developmental and environmental events should help define mechanistic connections among these factors in creating bimodal growth patterns.

Seasonal and diurnal trends in progressive isotope enrichment along needles in two pine species.

The Craig-Gordon type (C-G) leaf water isotope enrichment models assume a homogeneous distribution of enriched water across the leaf surface, despite observations that Δ18 O can become increasingly enriched from leaf base to tip. Datasets of this 'progressive isotope enrichment' are limited, precluding a comprehensive understanding of (a) the magnitude and variability of progressive isotope enrichment, and (b) how progressive enrichment impacts the accuracy of C-G leaf water model predictions. Here, we present observations of progressive enrichment in two conifer species that capture seasonal and diurnal variability in environmental conditions. We further examine which leaf water isotope models best capture the influence of progressive enrichment on bulk needle water Δ18 O. Observed progressive enrichment was large and equal in magnitude across both species. The magnitude of this effect fluctuated seasonally in concert with vapour pressure deficit, but was static in the face of diurnal cycles in meteorological conditions. Despite large progressive enrichment, three variants of the C-G model reasonably successfully predicted bulk needle Δ18 O. Our results thus suggest that the presence of progressive enrichment does not impact the predictive success of C-G models, and instead yields new insight regarding the physiological and anatomical mechanisms that cause progressive isotope enrichment.

Landscape-scale restoration minimizes tree growth vulnerability to 21st century drought in a dry forest.

Increasing aridity is a challenge for forest managers and reducing stand density to minimize competition is a recognized strategy to mitigate drought impacts on growth. In many dry forests, the most widespread and common forest management programs currently being implemented focus on restoration of historical stand structures, primarily to minimize fire risk and enhance watershed function. The implications of these restoration projects for drought vulnerability are not well understood. Here, we examined how planned restoration treatments in the Four Forests Restoration Initiative, the largest forest restoration project in the United States, would alter landscape-scale patterns of forest growth and drought vulnerability throughout the 21st century. Using drought-growth relationships developed within the landscape, we considered a suite of climate and treatment scenarios and estimated average forest growth and the proportion of years with extremely low growth as a measure of vulnerability to long-term decline. Climatic shifts projected for this landscape include higher temperatures and shifting seasonal precipitation that promotes lower soil moisture availability in the early growing season and greater hot-dry stress, conditions negatively associated with tree growth. However, drought severity and the magnitude of future growth declines were moderated by the thinning treatments. Compared to historical conditions, proportional growth in mid-century declines by ~40% if thinning ceases or continues at the status quo pace. By comparison, proportional growth declines by only 20% if the Four Forest Restoration Initiative treatments are fully implemented, and <10% if stands are thinned even more intensively than currently planned. Furthermore, restoration treatments resulted in dramatically fewer years with extremely low growth in the future, a recognized precursor to forest decline and eventual tree mortality. Benefits from density reduction for mitigating drought-induced growth declines are more apparent in mid-century and under RCP4.5 than under RCP8.5 at the end of the century. Future climate is inherently uncertain, and our results only reflect the climate projections from the representative suite of models examined. Nevertheless, these results indicate that forest restoration projects designed for other objectives also have substantial benefits for minimizing future drought vulnerability in dry forests and provide additional incentive to accelerate the pace of restoration.

Plant carbohydrate depletion impairs water relations and spreads via ectomycorrhizal networks.

Under prolonged drought and reduced photosynthesis, plants consume stored nonstructural carbohydrates (NSCs). Stored NSC depletion may impair the regulation of plant water balance, but the underlying mechanisms are poorly understood, and whether such mechanisms are independent of plant water deficit is not known. If so, carbon costs of fungal symbionts could indirectly influence plant drought tolerance through stored NSC depletion. We connected well-watered Pinus ponderosa seedling pairs via ectomycorrhizal (EM) networks where one seedling was shaded (D) and the other kept illuminated (LD) and compared responses to seedling pairs in full light (L). We measured plant NSCs, osmotic and water potential, and transfer of 13 CO2 through EM to explore mechanisms linking stored NSCs to plant water balance regulation and identify potential tradeoffs between plant water retention and EM fungi under carbon-limiting conditions. NSCs decreased from L to LD to D seedlings. Even without drought, NSC depletion impaired osmoregulation and turgor maintenance, both of which are critical for drought tolerance. Importantly, EM networks propagated NSC depletion and its negative effects on water retention from carbon stressed to nonstressed hosts. We demonstrate that NSC storage depletion influences turgor maintenance independently of plant water deficit and reveal carbon allocation tradeoffs between supporting fungal symbionts and retaining water.

Anatomical and hydraulic responses to desiccation in emergent conifer seedlings.

PREMISE: The young seedling life stage is critical for reforestation after disturbance and for species migration under climate change, yet little is known regarding their basic hydraulic function or vulnerability to drought. Here, we sought to characterize responses to desiccation including hydraulic vulnerability, xylem anatomical traits, and impacts on other stem tissues that contribute to hydraulic functioning. METHODS: Larix occidentalis, Pseudotsuga menziesii, and Pinus ponderosa (all ≤6 weeks old) were imaged using x-ray computed microtomography during desiccation to assess seedling biomechanical responses with concurrently measured hydraulic conductivity (ks ) and water potential (Ψ) to assess vulnerability to xylem embolism formation and other tissue damage. RESULTS: In non-stressed samples for all species, pith and cortical cells appeared circular and well hydrated, but they started to empty and deform with decreasing Ψ which resulted in cell tearing and eventual collapse. Despite the severity of this structural damage, the vascular cambium remained well hydrated even under the most severe drought. There were significant differences among species in vulnerability to xylem embolism formation, with 78% xylem embolism in L. occidentalis by Ψ of -2.1 MPa, but only 47.7% and 62.1% in P. ponderosa and P. menziesii at -4.27 and -6.73 MPa, respectively. CONCLUSIONS: Larix occidentalis seedlings appeared to be more susceptible to secondary xylem embolism compared to the other two species, but all three maintained hydration of the vascular cambium under severe stress, which could facilitate hydraulic recovery by regrowth of xylem when stress is relieved.

Variability and uncertainty in forest biomass estimates from the tree to landscape scale: the role of allometric equations.

BACKGROUND: Biomass maps are valuable tools for estimating forest carbon and forest planning. Individual-tree biomass estimates made using allometric equations are the foundation for these maps, yet the potentially-high uncertainty and bias associated with individual-tree estimates is commonly ignored in biomass map error. We developed allometric equations for lodgepole pine (Pinus contorta), ponderosa pine (P. ponderosa), and Douglas-fir (Pseudotsuga menziesii) in northern Colorado. Plot-level biomass estimates were combined with Landsat imagery and geomorphometric and climate layers to map aboveground tree biomass. We compared biomass estimates for individual trees, plots, and at the landscape-scale using our locally-developed allometric equations, nationwide equations applied across the U.S., and the Forest Inventory and Analysis Component Ratio Method (FIA-CRM). Total biomass map uncertainty was calculated by propagating errors from allometric equations and remote sensing model predictions. Two evaluation methods for the allometric equations were compared in the error propagation-errors calculated from the equation fit (equation-derived) and errors from an independent dataset of destructively-sampled trees (n = 285). RESULTS: Tree-scale error and bias of allometric equations varied dramatically between species, but local equations were generally most accurate. Depending on allometric equation and evaluation method, allometric uncertainty contributed 30-75% of total uncertainty, while remote sensing model prediction uncertainty contributed 25-70%. When using equation-derived allometric error, local equations had the lowest total uncertainty (root mean square error percent of the mean [% RMSE] = 50%). This is likely due to low-sample size (10-20 trees sampled per species) allometric equations and evaluation not representing true variability in tree growth forms. When independently evaluated, allometric uncertainty outsized remote sensing model prediction uncertainty. Biomass across the 1.56 million ha study area and uncertainties were similar for local (2.1 billion Mg; % RMSE = 97%) and nationwide (2.2 billion Mg;  % RMSE = 94%) equations, while FIA-CRM estimates were lower and more uncertain (1.5 billion Mg;  % RMSE = 165%). CONCLUSIONS: Allometric equations should be selected carefully since they drive substantial differences in bias and uncertainty. Biomass quantification efforts should consider contributions of allometric uncertainty to total uncertainty, at a minimum, and independently evaluate allometric equations when suitable data are available.