Responses of stomatal density and carbon isotope composition of sugar maple and yellow birch foliage to N, P and CaSiO3 fertilization.

Stomatal density, stomatal length and carbon isotope composition can all provide insights into environmental controls on photosynthesis and transpiration. Stomatal measurements can be time-consuming; it is therefore wise to consider efficient sampling schemes. Knowing the variance partitioning at different measurement levels (i.e., among stands, plots, trees, leaves and within leaves) can aid in making informed decisions around where to focus sampling effort. In this study, we explored the effects of nitrogen (N), phosphorus (P) and calcium silicate (CaSiO3) addition on stomatal density, length and carbon isotope composition (δ13C) of sugar maple (Acer saccharum Marsh.) and yellow birch (Betula alleghaniensis Britton). We observed a positive but small (8%) increase in stomatal density with P addition and an increase in δ13C with N and CaSiO3 addition in sugar maple, but we did not observe effects of nutrient addition on these characteristics in yellow birch. Variability was highest within leaves and among trees for stomatal density and highest among stomata for stomatal length. To reduce variability and increase chances of detecting treatment differences in stomatal density and length, future protocols should consider pretreatment and repeated measurements of trees over time or measure more trees per plot, increase the number of leaf impressions or standardize their locations, measure more stomata per image and ensure consistent light availability.

Sensitivity and threshold dynamics of Pinus strobus and Quercus spp. in response to experimental and naturally occurring severe droughts.

Increased drought frequency and severity are a pervasive global threat, yet the capacity of mesic temperate forests to maintain resilience in response to drought remains poorly understood. We deployed a throughfall removal experiment to simulate a once in a century drought in New Hampshire, USA, which coupled with the region-wide 2016 drought, intensified moisture stress beyond that experienced in the lifetimes of our study trees. To assess the sensitivity and threshold dynamics of two dominant northeastern tree genera (Quercus and Pinus), we monitored sap flux density (Js), leaf water potential and gas exchange, growth and intrinsic water-use efficiency (iWUE) for one pretreatment year (2015) and two treatment years (2016-17). Results showed that Js in pine (Pinus strobus L.) declined abruptly at a soil moisture threshold of 0.15 m3 m-3, whereas oak's (Quercus rubra L. and Quercus velutina Lam.) threshold was 0.11 m3 m-3-a finding consistent with pine's more isohydric strategy. Nevertheless, once oaks' moisture threshold was surpassed, Js declined abruptly, suggesting that while oaks are well adapted to moderate drought, they are highly susceptible to extreme drought. The radial growth reduction in response to the 2016 drought was more than twice as great for pine as for oaks (50 vs 18%, respectively). Despite relatively high precipitation in 2017, the oaks' growth continued to decline (low recovery), whereas pine showed neutral (treatment) or improved (control) growth. The iWUE increased in 2016 for both treatment and control pines, but only in treatment oaks. Notably, pines exhibited a significant linear relationship between iWUE and precipitation across years, whereas the oaks only showed a response during the driest conditions, further underscoring the different sensitivity thresholds for these species. Our results provide new insights into how interactions between temperate forest tree species' contrasting physiologies and soil moisture thresholds influence their responses and resilience to extreme drought.

Correcting tree-ring δ13C time series for tree-size effects in eight temperate tree species.

Stable carbon isotope ratios (δ13C) in tree rings have been widely used to study changes in intrinsic water-use efficiency (iWUE), sometimes with limited consideration of how C-isotope discrimination is affected by tree height and canopy position. Our goals were to quantify the relationships between tree size or tree microenvironment and wood δ13C for eight functionally diverse temperate tree species in northern New England and to better understand the physical and physiological mechanisms underlying these differences. We collected short increment cores in closed-canopy stands and analyzed δ13C in the most recent 5 years of growth. We also sampled saplings in both shaded and sun-exposed environments. In closed-canopy stands, we found strong tree-size effects on δ13C, with 3.7-7.2‰ of difference explained by linear regression vs height (0.11-0.28‰ m-1), which in some cases is substantially stronger than the effect reported in previous studies. However, open-grown saplings were often isotopically more similar to large codominant trees than to shade-grown saplings, indicating that light exposure contributes more to the physiological and isotopic differences between small and large trees than does height. We found that in closed-canopy forests, δ13C correlations with diameter at breast height were nonlinear but also strong, allowing a straightforward procedure to correct tree- or stand-scale δ13C-based iWUE chronologies for changing tree size. We demonstrate how to use such data to correct and interpret multi-decadal composite isotope chronologies in both shade-regenerated and open-grown tree cohorts, and we highlight the importance of understanding site history when interpreting δ13C time series.

Intensified vegetation water use under acid deposition.

Despite the important role vegetation plays in the global water cycle, the exact controls of vegetation water use, especially the role of soil biogeochemistry, remain elusive. In this study, we reveal a new mechanism of soil biogeochemical control of large-scale vegetation water use. Nitrate and sulfate deposition from fossil fuel burning have caused substantial soil acidification, leading to the leaching of soil base cations. Of these, calcium has a unique role in plant cells by regulating stomatal aperture, thus affecting vegetation water use. We hypothesized that the leaching of the soil calcium supply, induced by acid deposition, would increase large-scale vegetation water use. We present evidence from a long-term whole watershed acidification experiment demonstrating that the alteration of the soil calcium supply by acid deposition can significantly intensify vegetation water use (~10% increase in evapotranspiration) and deplete available soil water. These results are critical to understanding future water availability, biogeochemical cycles, and surface energy flux and to help reduce uncertainties in terrestrial biosphere models.

Phosphorus limitation of aboveground production in northern hardwood forests.

Forest productivity on glacially derived soils with weatherable phosphorus (P) is expected to be limited by nitrogen (N), according to theories of long-term ecosystem development. However, recent studies and model simulations based on resource optimization theory indicate that productivity can be co-limited by N and P. We conducted a full factorial N × P fertilization experiment in 13 northern hardwood forest stands of three age classes in central New Hampshire, USA, to test the hypothesis that forest productivity is co-limited by N and P. We also asked whether the response of productivity to N and P addition differs among species and whether differential species responses contribute to community-level co-limitation. Plots in each stand were fertilized with 30 kg N·ha-1 ·yr-1 , 10 kg P·ha-1 ·yr-1 , N + P, or neither nutrient (control) for four growing seasons. The productivity response to treatments was assessed using per-tree annual relative basal area increment (RBAI) as an index of growth. RBAI responded significantly to P (P = 0.02) but not to N (P = 0.73). However, evidence for P limitation was not uniform among stands. RBAI responded to P fertilization in mid-age (P = 0.02) and mature (P = 0.07) stands, each taken as a group, but was greatest in N-fertilized plots of two stands in these age classes, and there was no significant effect of P in the young stands. Both white birch (Betula papyrifera Marsh.) and beech (Fagus grandifolia Ehrh.) responded significantly to P; no species responded significantly to N. We did not find evidence for N and P co-limitation of tree growth. The response to N + P did not differ from that to P alone, and there was no significant N × P interaction (P = 0.68). Our P limitation results support neither the N limitation prediction of ecosystem theory nor the N and P co-limitation prediction of resource optimization theory, but could be a consequence of long-term anthropogenic N deposition in these forests. Inconsistencies in response to P suggest that successional status and variation in site conditions influence patterns of nutrient limitation and recycling across the northern hardwood forest landscape.

Temporal changes in soil C-N-P stoichiometry over the past 60 years across subtropical China.

Controlled experiments have shown that global changes decouple the biogeochemical cycles of carbon (C), nitrogen (N), and phosphorus (P), resulting in shifting stoichiometry that lies at the core of ecosystem functioning. However, the response of soil stoichiometry to global changes in natural ecosystems with different soil depths, vegetation types, and climate gradients remains poorly understood. Based on 2,736 observations along soil profiles of 0-150 cm depth from 1955 to 2016, we evaluated the temporal changes in soil C-N-P stoichiometry across subtropical China, where soils are P-impoverished, with diverse vegetation, soil, and parent material types and a wide range of climate gradients. We found a significant overall increase in soil total C concentration and a decrease in soil total P concentration, resulting in increasing soil C:P and N:P ratios during the past 60 years across all soil depths. Although average soil N concentration did not change, soil C:N increased in topsoil while decreasing in deeper soil. The temporal trends in soil C-N-P stoichiometry differed among vegetation, soil, parent material types, and spatial climate variations, with significantly increased C:P and N:P ratios for evergreen broadleaf forest and highly weathered Ultisols, and more pronounced temporal changes in soil C:N, N:P, and C:P ratios at low elevations. Our sensitivity analysis suggests that the temporal changes in soil stoichiometry resulted from elevated N deposition, rising atmospheric CO2 concentration and regional warming. Our findings revealed that the responses of soil C-N-P and stoichiometry to long-term global changes have occurred across the whole soil depth in subtropical China and the magnitudes of the changes in soil stoichiometry are dependent on vegetation types, soil types, and spatial climate variations.

Response of Quercus velutina growth and water use efficiency to climate variability and nitrogen fertilization in a temperate deciduous forest in the northeastern USA.

Nitrogen (N) deposition and changing climate patterns in the northeastern USA can influence forest productivity through effects on plant nutrient relations and water use. This study evaluates the combined effects of N fertilization, climate and rising atmospheric CO2on tree growth and ecophysiology in a temperate deciduous forest. Tree ring widths and stable carbon (δ(13)C) and oxygen (δ(18)O) isotopes were used to assess tree growth (basal area increment, BAI) and intrinsic water use efficiency (iWUE) ofQuercus velutinaLamb., the dominant tree species in a 20+ year N fertilization experiment at Harvard Forest (MA, USA). We found that fertilized trees exhibited a pronounced and sustained growth enhancement relative to control trees, with the low- and high-N treatments responding similarly. All treatments exhibited improved iWUE over the study period (1984-2011). Intrinsic water use efficiency trends in the control trees were primarily driven by changes in stomatal conductance, while a stimulation in photosynthesis, supported by an increase in foliar %N, contributed to enhancing iWUE in fertilized trees. All treatments were predominantly influenced by growing season vapor pressure deficit (VPD), with BAI responding most strongly to early season VPD and iWUE responding most strongly to late season VPD. Nitrogen fertilization increasedQ. velutinasensitivity to July temperature and precipitation. Combined, these results suggest that ambient N deposition in N-limited northeastern US forests has enhanced tree growth over the past 30 years, while rising ambient CO2has improved iWUE, with N fertilization and CO2having synergistic effects on iWUE.

Soil nitrogen affects phosphorus recycling: foliar resorption and plant-soil feedbacks in a northern hardwood forest.

Previous studies have attempted to link foliar resorption of nitrogen and phosphorus to their. respective availabilities in soil, with mixed results. Based on resource optimization theory, we hypothesized that the foliar resorption of one element could be driven by the availability of another element. We tested various measures of soil N and P as predictors of N and P resorption in six tree species in 18 plots across six stands at the Bartlett Experimental Forest, New Hampshire, USA. Phosphorus resorption efficiency (P < 0.01) and proficiency (P = 0.01) increased with soil N content. to 30 cm depth, suggesting that trees conserve P based on the availability of soil N. Phosphorus resorption also increased with soil P content, which is difficult to explain basdd on single-element limitation, butfollows from the correlation between soil N and soil P. The expected single-element relationships were evident only in the 0 horizon: P resorption was high where resin-available P was low in the Oe (P < 0.01 for efficiency, P < 0.001 for proficiency) and N resorption was high where potential N mineralization in the Oa was low (P < 0.01 for efficiency and 0.11 for proficiency). Since leaf litter is a principal source of N and P to the 0 horizon, low nutrient availability there could be a result rather than a cause of high resorption. The striking effect of soil N content on foliar P resorption is the first evidence of multiple-element control on nutrient resorption to be reported from an unmanipulated ecosystem.

The promise and peril of intensive-site-based ecological research: insights from the Hubbard Brook ecosystem study.

Ecological research is increasingly concentrated at particular locations or sites. This trend reflects a variety of advantages of intensive, site-based research, but also raises important questions about the nature of such spatially delimited research: how well does site based research represent broader areas, and does it constrain scientific discovery? We provide an overview of these issues with a particular focus on one prominent intensive research site: the Hubbard Brook Experimental Forest (HBEF), New Hampshire, USA. Among the key features of intensive sites are: long-term, archived data sets that provide a context for new discoveries and the elucidation of ecological mechanisms; the capacity to constrain inputs and parameters, and to validate models of complex ecological processes; and the intellectual cross-fertilization among disciplines in ecological and environmental sciences. The feasibility of scaling up ecological observations from intensive sites depends upon both the phenomenon of interest and the characteristics of the site. An evaluation of deviation metrics for the HBEF illustrates that, in some respects, including sensitivity and recovery of streams and trees from acid deposition, this site is representative of the Northern Forest region, of which HBEF is a part. However, the mountainous terrain and lack of significant agricultural legacy make the HBEF among the least disturbed sites in the Northern Forest region. Its relatively cool, wet climate contributes to high stream flow compared to other sites. These similarities and differences between the HBEF and the region can profoundly influence ecological patterns and processes and potentially limit the generality of observations at this and other intensive sites. Indeed, the difficulty of scaling up may be greatest for ecological phenomena that are sensitive to historical disturbance and that exhibit the greatest spatiotemporal variation, such as denitrification in soils and the dynamics of bird communities. Our research shows that end member sites for some processes often provide important insights into the behavior of inherently heterogeneous ecological processes. In the current era of rapid environmental and biological change, key ecological responses at intensive sites will reflect both specific local drivers and regional trends.

From missing source to missing sink: long-term changes in the nitrogen budget of a northern hardwood forest.

Biogeochemical monitoring for 45 years at the Hubbard Brook Experimental Forest in New Hampshire has revealed multiple surprises, seeming contradictions, and unresolved questions in the long-term record of ecosystem nitrogen dynamics. From 1965 to 1977, more N was accumulating in living biomass than was deposited from the atmosphere; the "missing" N source was attributed to biological fixation. Since 1992, biomass accumulation has been negligible or even negative, and streamwater export of dissolved inorganic N has decreased from ~4 to ~1 kg of N ha(-1) year(-1), despite chronically elevated atmospheric N deposition (~7 kg of N ha(-1) year(-1)) and predictions of N saturation. Here we show that the ecosystem has shifted to a net N sink, either storing or denitrifying ~8 kg of N ha(-1) year(-1). Repeated sampling over 25 years shows that the forest floor is not detectably accumulating N, but the C:N ratio is increasing. Mineral soil N has decreased nonsignificantly in recent decades, but the variability of these measurements prevents detection of a change of <700 kg of N ha(-1). Whether the excess N is accumulating in the ecosystem or lost through denitrification will be difficult to determine, but the distinction has important implications for the local ecosystem and global climate.