Season of drought affects growth, but not nonstructural carbohydrates dynamics, in Pinus taeda saplings.

In temperate evergreen conifers, growth occurs mostly in summer but photosynthesis precedes year-round; thus, nonstructural carbohydrates (NSC) increase in winter but decrease in summer. Given that mild drought reduces growth but not photosynthesis, a drought in summer should increase NSCs more than one in winter. However, the active regulation hypothesis suggests that to increase future drought resilience, plants might downregulate growth to increase NSCs after a winter drought even if NSCs do not increase during the drought. To test if so, potted Pinus taeda saplings (age $< 1$ yr) were subjected to six-month droughts in a greenhouse with one treatment receiving drought during winter (Sep-Mar), and another during summer (Mar-Sep). Both treatments were compared to a control. To measure dry biomass and NSCs, we harvested plants monthly following each drought, while to assess changes in growth rates, we measured height and diameter monthly. While we observed seasonal variation and an overall increase during the study, we found no drought-related changes in NSC dynamics; however, drought did reduce growth. Furthermore, drought in winter did reduce growth during the following summer, but the reduction was less than for a drought in summer. We conclude that the effect of drought on NSCs was too small to detect in our plants. While better control of soil water would have reduced a major source of uncertainty, plants with larger NSC reserves or more intense stress would also yield easier-to-detect effects. Although not definitive, our results suggest that water stress does not lead to dramatic changes in seasonal NSC dynamics in its aftermath, despite what one might expect under the active regulation hypothesis.

Modeling starch dynamics from seasonal variations of photosynthesis, growth, and respiration.

Nonstructural carbohydrates (NSCs) buffer differences in plant carbon supply (photosynthesis) and demand (respiration, growth, etc.) but the regulation of their dynamics remains unresolved. Seasonal variations in NSCs are well-documented, but differences in the time-average, amplitude, phase, and other characteristics across ecosystems and functional types lack explanation; furthermore, observed dynamics do not always match expectations. The failure to match observed and expected dynamics has stimulated debate on whether carbon supply or demand drives NSC dynamics. To gain insight into how carbon supply and demand drive seasonal NSC dynamics, we derive a simple model of NSC dynamics based on carbon mass balance and linearizing the NSC demand to determine how supply-driven and demand-driven seasonal NSC dynamics differ. We find that supply-driven and demand-driven dynamics yield distinct timings of seasonal extrema, and supply overrides demand when carbon supply is low in winter (e.g., at high latitudes). Our results also suggest that NSC dynamics often lag changes carbon mass balance. We also predict differences in NSC dynamics across mass, suggesting saplings are more dynamics and respond faster to the environment than mature trees. Our findings suggest substrate-dependent regulation with environmental variation is sufficient to generate complex NSC dynamics.

Tree crown injury from wildland fires: causes, measurement and ecological and physiological consequences.

The dead foliage of scorched crowns is one of the most conspicuous signatures of wildland fires. Globally, crown scorch from fires in savannas, woodlands and forests causes tree stress and death across diverse taxa. The term crown scorch, however, is inconsistently and ambiguously defined in the literature, causing confusion and conflicting interpretation of results. Furthermore, the underlying mechanisms causing foliage death from fire are poorly understood. The consequences of crown scorch - alterations in physiological, biogeochemical and ecological processes and ecosystem recovery pathways - remain largely unexamined. Most research on the topic assumes the mechanism of leaf and bud death is exposure to lethal air temperatures, with few direct measurements of lethal heating thresholds. Notable information gaps include how energy transfer injures and kills leaves and buds, how nutrients, carbohydrates, and hormones respond, and what physiological consequences lead to mortality. We clarify definitions to encourage use of unified terminology for foliage and bud necrosis resulting from fire. We review the current understanding of the physical mechanisms driving foliar injury, discuss the physiological responses, and explore novel ecological consequences of crown injury from fire. From these elements, we propose research needs for the increasingly interdisciplinary study of fire effects.

Xylem transport of root-derived CO2 caused a substantial underestimation of belowground respiration during a growing season.

Previous research has indicated that a potentially large portion of root-respired CO2 can move internally through tree xylem, but these reports are relatively scarce and have generally been limited to short observations. Our main objective was to provide a continuous estimate of the quantity and variability of root-respired CO2 that moves either internally through the xylem (FT ) or externally through the soil to the atmosphere (FS ) over most of a growing season. Nine trees were measured in a Populus deltoides stand for 129 days from early June to mid-October. We calculated FT as the product of sap flow and dissolved [CO2 ] in the xylem (i.e., [CO2 *]) and calculated FS using the [CO2 ] gradient method. During the study, stem and soil CO2 concentrations, temperature, and sap flow were measured continuously. We determined that FT accounted for 33% of daily total belowground CO2 flux (i.e., FS  + FT ; FB ) during our observation period that spanned most of a growing season. Cumulative daily FT was lower than FS 74% of the time, equivalent to FS 26% of the time, and never exceeded FS . One-third of the total CO2 released by belowground respiration over most of the growing season in this forest stand followed the FT pathway rather than diffusing into the soil. The magnitude of FT indicates that measurements of FS alone substantially underestimate total belowground respiration in some forest ecosystems by systematically underestimating belowground autotrophic respiration. The variability in FT observed during the growing season demonstrated the importance of making long-term, high-frequency measurements of different flux pathways to better understand physiological and ecological processes and their implications to global change.

Integration of ecosystem science into radioecology: A consensus perspective.

In the Fall of 2016 a workshop was held which brought together over 50 scientists from the ecological and radiological fields to discuss feasibility and challenges of reintegrating ecosystem science into radioecology. There is a growing desire to incorporate attributes of ecosystem science into radiological risk assessment and radioecological research more generally, fueled by recent advances in quantification of emergent ecosystem attributes and the desire to accurately reflect impacts of radiological stressors upon ecosystem function. This paper is a synthesis of the discussions and consensus of the workshop participant's responses to three primary questions, which were: 1) How can ecosystem science support radiological risk assessment? 2) What ecosystem level endpoints potentially could be used for radiological risk assessment? and 3) What inference strategies and associated methods would be most appropriate to assess the effects of radionuclides on ecosystem structure and function? The consensus of the participants was that ecosystem science can and should support radiological risk assessment through the incorporation of quantitative metrics that reflect ecosystem functions which are sensitive to radiological contaminants. The participants also agreed that many such endpoints exit or are thought to exit and while many are used in ecological risk assessment currently, additional data need to be collected that link the causal mechanisms of radiological exposure to these endpoints. Finally, the participants agreed that radiological risk assessments must be designed and informed by rigorous statistical frameworks capable of revealing the causal inference tying radiological exposure to the endpoints selected for measurement.

Stand development and other intrinsic factors largely control fine-root dynamics with only subtle modifications from resource availability.

Forest productivity depends on resource acquisition by ephemeral roots and leaves. A combination of intrinsic and environmental factors influences ephemeral organs; however, difficulties in studying belowground organs impede mechanistic understanding of fine-root production and turnover. To quantify factors controlling fine-root dynamics, we grew a deciduous hardwood (Populus deltoides Bartr.) and an evergreen conifer (Pinus taeda L.) with distinct soil moisture and nutrient availability treatments. We monitored fine-root dynamics with minirhizotrons for 6 years during early stand development and expressed results on a root length, biomass and mortality-risk basis. Stand development and other intrinsic factors consistently influenced both species in the same direction and by similar magnitude. Live-root length increased to a peak during establishment and slowly declined after roots of neighboring trees overlapped. Root longevity was highest during establishment and decreased thereafter. Root longevity consistently increased with depth of appearance and initial root diameter. Season of appearance affected root longevity in the following order: spring > summer > fall > winter. The influence of soil resource availability on fine-root dynamics was inconsistent between species, and ranked below that of rooting depth, initial diameter, stand development and phenology. Fine-root biomass either increased or was unaffected by greater resource availability. Fine-root production and live root length decreased with irrigation for both species, and increased with fertilization only for poplar. Fine-root mortality risk both increased and decreased depending on species and amendment treatment. Differing responses to soil moisture and nutrient availability between species suggests we should carefully evaluate generalizations about the response of fine-root dynamics to resource availability. While attempting to describe and explain carbon allocation to fine-root production and turnover, modelers and physiologists should first consider consistent patterns of allocation caused by different depth, diameter, stand development, phenology and species before considering allocation due to soil resource availability.

Stored root carbohydrates can maintain root respiration for extended periods.

Tight coupling between below-ground autotrophic respiration and the availability of recently assimilated carbon (C) has become a paradigm in the ecophysiological literature. Here, we show that stored carbohydrates can decouple respiration from assimilation for prolonged periods by mobilizing reserves from transport roots to absorptive roots. We permanently disrupted the below-ground transfer of recently assimilated C using stem girdling and root trenching and measured soil CO2 efflux for over 1 yr in longleaf pine (Pinus palustris), a species that has large reserves of stored carbohydrates in roots. Soil CO2 efflux was not influenced by girdling or trenching through the 14-month observation period. Stored carbohydrate concentrations in absorptive roots were not affected by the disrupted supply of current photosynthate for over 1 yr; however, carbohydrate concentrations in transport roots decreased. Our results indicate that root respiration can be decoupled from recent canopy assimilation and that stored carbohydrates can be mobilized from transport roots to absorptive roots to maintain respiration for over 1 yr. This refines the current paradigm that canopy assimilation and below-ground respiration are tightly coupled and provides evidence of the mechanism and dynamics responsible for decoupling the above- and below-ground processes.

A quantitative method for analyzing glycome profiles of plant cell walls.

Glycome profiling allows for the characterization of plant cell wall ultrastructure via sequential extractions and subsequent detection of specific epitopes with a suite of glycan-specific monoclonal antibodies (mAbs). The data are often viewed as the amount of materials recovered and coinciding colored heatmaps of mAb binding are generated. Interpretation of these data can be considered qualitative in nature as it depends on detecting subtle visual differences in antibody binding strength. Here, we report a mixed model-based quantitative approach for glycome profile analyses, which accounts for the amount of materials recovered and displays the normalized values in revised heatmaps and statistical heatmaps depicting significant differences. The utility of this methodology was demonstrated on a previously published dataset investigating the effects of moisture stress on the roots and needles of Pinus taeda. An annotated R script for the quantitative methodology is included to allow future studies to utilize the same approach.

Cell Wall Ultrastructure of Stem Wood, Roots, and Needles of a Conifer Varies in Response to Moisture Availability.

The composition, integrity, and architecture of the macromolecular matrix of cell walls, collectively referred to as cell wall ultrastructure, exhibits variation across species and organs and among cell types within organs. Indirect approaches have suggested that modifications to cell wall ultrastructure occur in response to abiotic stress; however, modifications have not been directly observed. Glycome profiling was used to study cell wall ultrastructure by examining variation in composition and extractability of non-cellulosic glycans in cell walls of stem wood, roots, and needles of loblolly pine saplings exposed to high and low soil moisture. Soil moisture influenced physiological processes and the overall composition and extractability of cell wall components differed as a function of soil moisture treatments. The strongest response of cell wall ultrastructure to soil moisture was increased extractability of pectic backbone epitopes in the low soil moisture treatment. The higher abundance of these pectic backbone epitopes in the oxalate extract indicate that the loosening of cell wall pectic components could be associated with the release of pectic signals as a stress response. The increased extractability of pectic backbone epitopes in response to low soil moisture availability was more pronounced in stem wood than in roots or needles. Additional responses to low soil moisture availability were observed in lignin-associated carbohydrates released in chlorite extracts of stem wood, including an increased abundance of pectic arabinogalactan epitopes. Overall, these results indicate that cell walls of loblolly pine organs undergo changes in their ultrastructural composition and extractability as a response to soil moisture availability and that cell walls of the stem wood are more responsive to low soil moisture availability compared to cell walls of roots and needles. To our knowledge, this is the first direct evidence, delineated by glycomic analyses, that abiotic stress affects cell wall ultrastructure. This study is also unique in that glycome profiling of pine needles has never before been reported.

Temporal and spatial patterns of internal and external stem CO2 fluxes in a sub-Mediterranean oak.

To accurately estimate stem respiration (RS), measurements of both carbon dioxide (CO2) efflux to the atmosphere (EA) and internal CO2 flux through xylem (FT) are needed because xylem sap transports respired CO2 upward. However, reports of seasonal dynamics of FT and EA are scarce and no studies exist in Mediterranean species under drought stress conditions. Internal and external CO2 fluxes at three stem heights, together with radial stem growth, temperature, sap flow and shoot water potential, were measured in Quercus pyrenaica Willd. in four measurement campaigns during one growing season. Substantial daytime depressions in temperature-normalized EA were observed throughout the experiment, including prior to budburst, indicating that diel hysteresis between stem temperature and EA cannot be uniquely ascribed to diversion of CO2 in the transpiration stream. Low internal [CO2] (<0.5%) resulted in low contributions of FT to RS throughout the growing season, and RS was mainly explained by EA (>90%). Internal [CO2] was found to vary vertically along the stems. Seasonality in resistance to radial CO2 diffusion was related to shoot water potential. The low internal [CO2] and FT observed in our study may result from the downregulation of xylem respiration in response to a legacy of coppicing as well as high radial diffusion of CO2 through cambium, phloem and bark tissues, which was related to low water content of stems. Long-term studies analyzing temporal and spatial variation in internal and external CO2 fluxes and their interactions are needed to mechanistically understand and model respiration of woody tissues.

Global patterns and predictors of stem CO2 efflux in forest ecosystems.

Stem CO2 efflux (ES) plays an important role in the carbon balance of forest ecosystems. However, its primary controls at the global scale are poorly understood and observation-based global estimates are lacking. We synthesized data from 121 published studies across global forest ecosystems and examined the relationships between annual ES and biotic and abiotic factors at individual, biome, and global scales, and developed a global gridded estimate of annual ES . We tested the following hypotheses: (1) Leaf area index (LAI) will be highly correlated with annual ES at biome and global scales; (2) there will be parallel patterns in stem and root CO2 effluxes (RA) in all forests; (3) annual ES will decline with forest age; and (4) LAI coupled with mean annual temperature (MAT) and mean annual precipitation (MAP) will be sufficient to predict annual ES across forests in different regions. Positive linear relationships were found between ES and LAI, as well as gross primary production (GPP), net primary production (NPP), wood NPP, soil CO2 efflux (RS), and RA . Annual ES was correlated with RA in temperate forests after controlling for GPP and MAT, suggesting other additional factors contributed to the relationship. Annual ES tended to decrease with stand age. Leaf area index, MAT and MAP, predicted 74% of variation in ES at global scales. Our statistical model estimated a global annual ES of 6.7 ± 1.1 Pg C yr(-1) over the period of 2000-2012 with little interannual variability. Modeled mean annual ES was 71 ± 43, 270 ± 103, and 420 ± 134 g C m(2) yr(-1) for boreal, temperate, and tropical forests, respectively. We recommend that future studies report ES at a standardized constant temperature, incorporate more manipulative treatments, such as fertilization and drought, and whenever possible, simultaneously measure both aboveground and belowground CO2 fluxes.

Poplar saplings exposed to recurring temperature shifts of different amplitude exhibit differences in leaf gas exchange and growth despite equal mean temperature.

Most investigations of plant responses to changes in temperature have focused on a constant increase in mean day/night temperature without considering how differences in temperature cycles can affect physiological processes and growth. To test the effects of changes in growth temperature on foliar carbon balance and plant growth, we repeatedly exposed poplar saplings (Populus deltoides × nigra) to temperature cycles consisting of 5 days of a moderate (M, +5 °C) or extreme (E, +10 °C) increase in temperature followed by 5 days of a moderate (M, -5 °C) or extreme (E, -10 °C) decrease in temperature, with respect to a control treatment (C, 23.4 °C). The temperature treatments had the same mean temperature over each warm and cool cycle and over the entire study. Our goal was to examine the influence of recurring temperature shifts on growth. Net photosynthesis (A) was relatively insensitive to changes in growth temperature (from 20 to 35 °C), suggesting a broad range of optimum temperature for photosynthesis. Leaf respiration (R) exhibited substantial acclimation to temperature, having nearly the same rate at 13 °C as at 33 °C. There was no evidence that preconditioning through temperature cycles affected the response of A or R to treatment temperature fluctuations. Averaged across the complete warm/cool temperature cycle, the A : R ratio did not differ among the temperature treatments. While foliar carbon balance was not affected, the temperature treatments significantly affected growth. Whole-plant biomass was 1.5 times greater in the M treatment relative to the C treatment. Carbon allocation was also affected with shoot volume and biomass greater in the M and E treatments than in the C treatment. Our findings indicate that temperature fluctuations can have important effects on growth, though there were few effects on leaf gas exchange, and can help explain differences in growth that are not correlated with mean growth temperature.

Stem girdling affects the quantity of CO2 transported in xylem as well as CO2 efflux from soil.

There is recent clear evidence that an important fraction of root-respired CO2 is transported upward in the transpiration stream in tree stems rather than fluxing to the soil. In this study, we aimed to quantify the contribution of root-respired CO2 to both soil CO2 efflux and xylem CO2 transport by manipulating the autotrophic component of belowground respiration. We compared soil CO2 efflux and the flux of root-respired CO2 transported in the transpiration stream in girdled and nongirdled 9-yr-old oak trees (Quercus robur) to assess the impact of a change in the autotrophic component of belowground respiration on both CO2 fluxes. Stem girdling decreased xylem CO2 concentration, indicating that belowground respiration contributes to the aboveground transport of internal CO2 . Girdling also decreased soil CO2 efflux. These results confirmed that root respiration contributes to xylem CO2 transport and that failure to account for this flux results in inaccurate estimates of belowground respiration when efflux-based methods are used. This research adds to the growing body of evidence that efflux-based measurements of belowground respiration underestimate autotrophic contributions.

Assimilation of xylem-transported CO2 is dependent on transpiration rate but is small relative to atmospheric fixation.

The effect of transpiration rate on internal assimilation of CO2 released from respiring cells has not previously been quantified. In this study, detached branches of Populus deltoides were allowed to take up (13)CO2-labelled solution at either high (high label, HL) or low (low label, LL) (13)CO2 concentrations. The uptake of the (13)CO2 label served as a proxy for the internal transport of respired CO2, whilst the transpiration rate was manipulated at the leaf level by altering the vapour pressure deficit (VPD) of the air. Simultaneously, leaf gas exchange was measured, allowing comparison of internal CO2 assimilation with that assimilated from the atmosphere. Subsequent (13)C analysis of branch and leaf tissues revealed that woody tissues assimilated more label under high VPD, corresponding to higher transpiration, than under low VPD. More (13)C was assimilated in leaf tissue than in woody tissue under the HL treatment, whereas more (13)C was assimilated in woody tissue than in leaf tissue under the LL treatment. The ratio of (13)CO2 assimilated from the internal source to CO2 assimilated from the atmosphere was highest for the branches under the HL and high VPD treatment, but was relatively small regardless of VPD×label treatment combination (up to 1.9%). These results showed that assimilation of internal CO2 is highly dependent on the rate of transpiration and xylem sap [CO2]. Therefore, it can be expected that the relative contribution of internal CO2 recycling to tree carbon gain is strongly dependent on factors controlling transpiration, respiration, and photosynthesis.

Internal recycling of respired CO 2 may be important for plant functioning under changing climate regimes.

Recent studies have provided evidence of a large flux of root-respired CO 2 in the transpiration stream of trees. In our study, we investigated the potential impact of this internal CO 2 transport on aboveground carbon assimilation and CO 2 efflux. To trace the transport of root-respired CO 2, we infused a (13)C label at the stem base of field-grown Populus deltoides Bartr. ex. Marsh trees. The (13)C label was transported to the top of the stem and throughout the crown via the transpiration stream. Up to 17% of the (13)C label was assimilated by chlorophyll-containing tissues. Our results provide evidence of a mechanism for recycling respired CO 2 within trees. Such a mechanism may have important implications for how plants cope with predicted increases in intensity and frequency of droughts. Here, we speculate on the potential significance of this recycling mechanism within the context of plant responses to climate change and plants currently inhabiting arid environments.

Transport of root-respired CO2 via the transpiration stream affects aboveground carbon assimilation and CO2 efflux in trees.

Upward transport of CO2 via the transpiration stream from belowground to aboveground tissues occurs in tree stems. Despite potentially important implications for our understanding of plant physiology, the fate of internally transported CO2 derived from autotrophic respiratory processes remains unclear. We infused a ¹³CO2-labeled aqueous solution into the base of 7-yr-old field-grown eastern cottonwood (Populus deltoides) trees to investigate the effect of xylem-transported CO2 derived from the root system on aboveground carbon assimilation and CO2 efflux. The ¹³C label was transported internally and detected throughout the tree. Up to 17% of the infused label was assimilated, while the remainder diffused to the atmosphere via stem and branch efflux. The largest amount of assimilated ¹³C was found in branch woody tissues, while only a small quantity was assimilated in the foliage. Petioles were more highly enriched in ¹³C than other leaf tissues. Our results confirm a recycling pathway for respired CO2 and indicate that internal transport of CO2 from the root system may confound the interpretation of efflux-based estimates of woody tissue respiration and patterns of carbohydrate allocation.

Root-derived CO(2) efflux via xylem stream rivals soil CO(2) efflux.

Respiration consumes a large portion of annual gross primary productivity in forest ecosystems and is dominated by belowground metabolism. Here, we present evidence of a previously unaccounted for internal CO(2) flux of large magnitude from tree roots through stems. If this pattern is shown to persist over time and in other forests, it suggests that belowground respiration has been grossly underestimated. Using an experimental Populus deltoides plantation as a model system, we tested the hypothesis that a substantial portion of the CO(2) released from belowground autotrophic respiration remains within tree root systems and is transported aboveground through the xylem stream rather than diffusing into the soil atmosphere. On a daily basis, the amount of CO(2) that moved upward from the root system into the stem via the xylem stream (0.26 mol CO(2) m(-2) d(-1)) rivalled that which diffused from the soil surface to the atmosphere (0.27 mol CO(2) m(-2) d(-1)). We estimated that twice the amount of CO(2) derived from belowground autotrophic respiration entered the xylem stream as diffused into the soil environment. Our observations indicate that belowground autotrophic respiration consumes substantially more carbohydrates than previously recognized and challenge the paradigm that all root-respired CO(2) diffuses into the soil atmosphere.