Range-wide variations in common milkweed traits and their effect on monarch larvae.

PREMISE: Leaf economic spectrum (LES) theory has historically been employed to inform vegetation models of ecosystem processes, but largely neglects intraspecific variation and biotic interactions. We attempt to integrate across environment-plant trait-herbivore interactions within a species at a range-wide scale. METHODS: We measured traits in 53 populations spanning the range of common milkweed (Asclepias syriaca) and used a common garden to determine the role of environment in driving patterns of intraspecific variation. We used a feeding trial to determine the role of plant traits in monarch (Danaus plexippus) larval development. RESULTS: Trait-trait relationships largely followed interspecific patterns in LES theory and persisted in a common garden when individual traits change. Common milkweed showed intraspecific variation and biogeographic clines in traits. Clines did not persist in a common garden. Larvae ate more and grew larger when fed plants with more nitrogen. A longitudinal environmental gradient in precipitation corresponded to a resource gradient in plant nitrogen, which produces a gradient in larval performance. CONCLUSIONS: Biogeographic patterns in common milkweed traits can sometimes be predicted from LES, are largely driven by environmental conditions, and have consequences for monarch larval performance. Changes to nutrient dynamics of landscapes with common milkweed could potentially influence monarch population dynamics. We show how biogeographic trends in intraspecific variation can influence key ecological interactions, especially in common species with large distributions.

Remote spectral detection of biodiversity effects on forest biomass.

Quantifying how biodiversity affects ecosystem functions through time over large spatial extents is needed for meeting global biodiversity goals yet is infeasible with field-based approaches alone. Imaging spectroscopy is a tool with potential to help address this challenge. Here, we demonstrate a spectral approach to assess biodiversity effects in young forests that provides insight into its underlying drivers. Using airborne imaging of a tree-diversity experiment, spectral differences among stands enabled us to quantify net biodiversity effects on stem biomass and canopy nitrogen. By subsequently partitioning these effects, we reveal how distinct processes contribute to diversity-induced differences in stand-level spectra, chemistry and biomass. Across stands, biomass overyielding was best explained by species with greater leaf nitrogen dominating upper canopies in mixtures, rather than intraspecific shifts in canopy structure or chemistry. Remote imaging spectroscopy may help to detect the form and drivers of biodiversity-ecosystem function relationships across space and time, advancing the capacity to monitor and manage Earth's ecosystems.

Spectral Phenotyping of Physiological and Anatomical Leaf Traits Related with Maize Water Status.

Advancements in phenotyping techniques capable of rapidly and nondestructively detecting impacts of drought on crops are necessary to meet the 21st-century challenge of food security. Here, we describe the use of hyperspectral reflectance to predict variation in physiological and anatomical leaf traits related with water status under varying water availability in six maize (Zea mays) hybrids that differ in yield stability under drought. We also assessed relationships among traits and collections of traits with yield stability. Measurements were collected in both greenhouse and field environments, with plants exposed to different levels of water stress or to natural water availability, respectively. Leaf spectral measurements were paired with a number of physiological and anatomical reference measurements, and predictive spectral models were constructed using a partial least-squares regression approach. All traits were relatively well predicted by spectroscopic models, with external validation (i.e. by applying partial least-squares regression coefficients on a dataset distinct from the one used for calibration) goodness-of-fit (R 2 ) ranging from 0.37 to 0.89 and normalized error ranging from 12% to 21%. Correlations between reference and predicted data were statistically similar for both greenhouse and field data. Our findings highlight the capability of vegetation spectroscopy to rapidly and nondestructively identify a number of foliar functional traits affected by drought that can be used as indicators of plant water status. Although we did not detect trait coordination with yield stability in the hybrids used in this study, expanding the range of functional traits estimated by hyperspectral data can help improve trait-based breeding approaches.

Fidelity and Timing of Spotted Lanternfly (Hemiptera: Fulgoridae) Attack Patterns on Ornamental Trees in the Suburban Landscape.

Invasive herbivores can have dramatic impacts in new environments by altering landscape composition, displacing natives, and causing plant decline and mortality. One of the most recent invasive insects in the United States, the spotted lanternfly (Lycorma delicatula), has the potential to cause substantial economic and environmental impacts in agriculture and forestry. Spotted lanternfly exhibits a broad host range, yet reports of late-season movement from the surrounding landscapes onto select tree species in suburban environments have been reported. In this study, we aimed to evaluate the fidelity of spotted lanternfly attack on specific, individual trees within the same species during this movement period. In 2018 and 2019, we observed that individual red (Acer rubrum L. [Sapindales: Sapindaceae]) and silver maple (Acer saccharinum L. [Sapindales: Sapindaceae]) trees were preferentially attacked over other nearby trees of the same species. Foliar elemental composition was a good predictor of spotted lanternfly attack numbers, indicating that individual variation in nutrients may influence spotted lanternfly attraction to and/or retention on maple trees. Our data also confirm reports of late-season movement from surrounding landscapes throughout autumn. Collectively, our results show that spotted lanternfly exhibits some fidelity to particular trees in the landscape during this movement period. While other potential mechanisms also contribute to host plant selection by spotted lanternfly, our data show that host nutritional profiles influence spotted lanternfly infestation of suburban trees at the landscape scale. Our data establish that late-season infestations of suburban trees by spotted lanternfly occurred and that variation in host quality should be further considered in the management of this invasive insect pest.

Foliar functional traits from imaging spectroscopy across biomes in eastern North America.

Foliar functional traits are widely used to characterize leaf and canopy properties that drive ecosystem processes and to infer physiological processes in Earth system models. Imaging spectroscopy provides great potential to map foliar traits to characterize continuous functional variation and diversity, but few studies have demonstrated consistent methods for mapping multiple traits across biomes. With airborne imaging spectroscopy data and field data from 19 sites, we developed trait models using partial least squares regression, and mapped 26 foliar traits in seven NEON (National Ecological Observatory Network) ecoregions (domains) including temperate and subtropical forests and grasslands of eastern North America. Model validation accuracy varied among traits (normalized root mean squared error, 9.1-19.4%; coefficient of determination, 0.28-0.82), with phenolic concentration, leaf mass per area and equivalent water thickness performing best across domains. Across all trait maps, 90% of vegetated pixels had reasonable values for one trait, and 28-81% provided high confidence for multiple traits concurrently. Maps of 26 traits and their uncertainties for eastern US NEON sites are available for download, and are being expanded to the western United States and tundra/boreal zone. These data enable better understanding of trait variations and relationships over large areas, calibration of ecosystem models, and assessment of continental-scale functional diversity.

Water stress rather than N addition mitigates impacts of elevated O3 on foliar chemical profiles in poplar saplings.

Tropospheric ozone (O3) pollution can alter tree chemical profiles, and in turn, affect forest ecosystem function. However, the magnitude of these effects may be modified by variations in soil water and nutrient availability, which makes it difficult to predict the impacts of O3 in reality. Here we assessed the effects of elevated O3 alone, and in combination with soil water deficit and N addition, on the phytochemical composition of hybrid poplar (Populus deltoides cv. '55/56' × P. deltoides cv. 'Imperial'). Potted trees were grown in open-top chambers (OTCs) under either charcoal-filtered air or elevated O3 (non-filtered air +40 ppb of O3), and trees within each OTC were grown with four combinations of water (well-watered or water deficit) and nitrogen (with or without N addition) levels. We found that elevated O3 alone stimulated the accumulation of foliar nitrogen, soluble sugar, and lignin while inhibiting the accumulation of starch, but had limited impacts on condensed tannins and salicinoids in poplar saplings. Graphical vector analysis revealed that these changes in concentrations of nitrogen, starch and lignin were due largely to altered metabolic processes, while increased soluble sugar concentration related mainly to decreased leaf biomass in most cases. The effects of O3 on poplar foliar chemical profiles depended on soil water, but not soil N, availability. Specifically, O3-mediated changes in carbohydrates and lignin were mitigated by decreased soil water content. Taken together, these results suggested that nitrogen acquisition, carbohydrates mobilization and lignification play a role in poplar tolerance to O3. Moreover, the impacts of elevated O3 on phytochemistry of poplar leaves can be context-dependent, with potential consequences for ecosystem processes under future global change scenarios. Our results highlight the needs to consider multi-factors environments to optimize the management of plantations under changing environments.

Spectral characterization of wheat functional trait responses to Hessian fly: Mechanisms for trait-based resistance.

Insect herbivores can manipulate host plants to inhibit defenses. Insects that induce plant galls are excellent examples of these interactions. The Hessian fly (HF, Mayetiola destructor) is a destructive pest of wheat (Triticum spp.) that occurs in nearly all wheat producing globally. Under compatible interactions (i.e., successful HF establishment), HF larvae alter host tissue physiology and morphology for their benefit, manifesting as the development of plant nutritive tissue that feeds the larva and ceases plant cell division and elongation. Under incompatible interactions (i.e., unsuccessful HF establishment), plants respond to larval feeding by killing the larva, permitting normal plant development. We used reflectance spectroscopy to characterize whole-plant functional trait responses during both compatible and incompatible interactions and related these findings with morphological and gene expression observations from earlier studies. Spectral models successfully characterized wheat foliar traits, with mean goodness of fit statistics of 0.84, 0.85, 0.94, and 0.69 and percent root mean square errors of 22, 10, 6, and 20%, respectively, for nitrogen and carbon concentrations, leaf mass per area, and total phenolic content. We found that larvae capable of generating compatible interactions successfully manipulated host plant chemical and morphological composition to create a more hospitable environment. Incompatible interactions resulted in lower host plant nutritional quality, thicker leaves, and higher phenolic levels. Spectral measurements successfully characterized wheat responses to compatible and incompatible interactions, providing an excellent example of the utility of Spectral phenotyping in quantifying responses of specific plant functional traits associated with insect resistance.

The demographic effects of functional traits: an integral projection model approach reveals population-level consequences of reproduction-defence trade-offs.

Quantitatively linking individual variation in functional traits to demography is a necessary step to advance our understanding of trait-based ecological processes. We constructed a population model for Asclepias syriaca to identify how functional traits affect vital rates and population growth and whether trade-offs in chemical defence and demography alter population growth. Plants with higher foliar cardenolides had lower fibre, cellulose and lignin levels, as well as decreased sexual and clonal reproduction. Average cardenolide concentrations had the strongest effect on population growth. In both the sexual and clonal pathway, the trade-off between reproduction and defence affected population growth. We found that both increasing the mean of the distribution of individual plant values for cardenolides and herbivory decreased population growth. However, increasing the variance in both defence and herbivory increased population growth. Functional traits can impact population growth and quantifying individual-level variation in traits should be included in assessments of population-level processes.

Reflectance spectroscopy: a novel approach to better understand and monitor the impact of air pollution on Mediterranean plants.

The Mediterranean basin can be considered a hot spot not only in terms of climate change (CC) but also for air quality. Assessing the impact of CC and air pollution on ecosystem functions is a challenging task, and adequate monitoring techniques are needed. This paper summarizes the present knowledge on the use of reflectance spectroscopy for the evaluation of the effects of air pollution on plants. First, the history of this technique is outlined. Next, we describe the vegetation reflectance spectrum, how it can be scaled from leaf to landscape levels, what information it contains, and how it can be exploited to understand plant and ecosystem functions. Finally, we review the literature concerning this topic, with special attention to Mediterranean air pollutants, showing the increasing interest in this technique. The ability of spectroscopy to detect the influence of air pollution on plant function of all major and minor Mediterranean pollutants has been evaluated, and ozone and its interaction with other gases (carbon dioxide, nitrogen oxides, and sulfur dioxide) have been the most studied. In the recent years, novel air pollutants, such as particulate matter, nitrogen deposition, and heavy metals, have drawn attention. Although various vegetation types have been studied, few of these species are representative of the Mediterranean environment. Thus, major emphasis should be placed on using vegetation spectroscopy for better understanding and monitoring the impact of air pollution on Mediterranean plants in the CC era.

Using foliar spectral properties to assess the effects of drought on plant water potential.

Drought frequency is predicted to increase in future environments. Leaf water potential (ΨLW) is commonly used to evaluate plant water status, but traditional measurements can be logistically difficult and require destructive sampling. We used reflectance spectroscopy to characterize variation in ΨLW of Quercus oleoides Schltdl. & Cham. under differential water availability and tested the ability to predict pre-dawn ΨLW (PDΨLW) using spectral data collected hours after pressure chamber measurements on dark-acclimated leaves. ΨLW was measured with a Scholander pressure chamber. Leaf reflectance was collected at one or both of two time points: immediately (ΨLW) and ~5 h after pressure chamber measurements (PDΨLW). Predictive models were constructed using partial least-squares regression. Model performance was evaluated using coefficient of determination (R2), root-mean-square error (RMSE), bias, and the percent RMSE of the data range (%RMSE). ΨLW and PDΨLW were well predicted using spectroscopic models and successfully estimated a wide variation in ΨLW (light- or dark-acclimated leaves) as well as PDΨLW (dark-acclimated leaves only). Mean ΨLWR2, RMSE and bias values were 0.65, 0.51 MPa and 0.09, respectively, with a %RMSE between 8% and 20%, while mean PDΨLWR2, RMSE and bias values were 0.60, 0.44 MPa and 0.01, respectively, with a %RMSE between 9% and 20%. Estimates of PDΨLW produced similar statistical outcomes when analyzing treatment effects on PDΨLW as those found using reference pressure chamber measurements. These findings highlight a promising approach to evaluate plant responses to environmental change by providing rapid measurements that can be used to estimate plant water status as well as demonstrating that spectroscopic measurements can be used as a surrogate for standard, reference measurements in a statistical framework.

Vernal freeze damage and genetic variation alter tree growth, chemistry, and insect interactions.

Anticipated consequences of climate change in temperate regions include early spring warmup punctuated by intermittent hard freezes. Warm weather accelerates leaf flush in perennial woody species, potentially exposing vulnerable young tissues to damaging frosts. We employed a 2 × 6 randomized factorial design to examine how the interplay of vernal (springtime) freeze damage and genetic variation in a hardwood species (Populus tremuloides) influences tree growth, phytochemistry, and interactions with an insect herbivore (Chaitophorus stevensis). Acute effects of freezing included defoliation and mortality. Surviving trees exhibited reduced growth and altered biomass distribution. Reflushed leaves on these trees had lower mass per area, lower lignin concentrations, and higher nitrogen concentrations, altered chemical defence profiles, and supported faster aphid population growth. Many effects varied among plant genotypes and were related with herbivore performance. This study suggests that a single damaging vernal freeze event can alter tree-insect interactions through effects on plant growth and chemistry. Differential responses of various genotypes to freeze damage suggest that more frequent vernal freeze events could also influence natural selection, favouring trees with greater freeze hardiness, and more resistance or tolerance to herbivores following damage.

Effects of Elevated Atmospheric Carbon Dioxide and Tropospheric Ozone on Phytochemical Composition of Trembling Aspen ( Populus tremuloides ) and Paper Birch ( Betula papyrifera ).

Anthropogenic activities are altering levels of atmospheric carbon dioxide (CO2) and tropospheric ozone (O3). These changes can alter phytochemistry, and in turn, influence ecosystem processes. We assessed the individual and combined effects of elevated CO2 and O3 on the phytochemical composition of two tree species common to early successional, northern temperate forests. Trembling aspen (Populus tremuloides) and paper birch (Betula papyrifera) were grown at the Aspen FACE (Free-Air Carbon dioxide and ozone Enrichment) facility under four combinations of ambient and elevated CO2 and O3. We measured, over three years (2006-08), the effects of CO2 and O3 on a suite of foliar traits known to influence forest functioning. Elevated CO2 had minimal effect on foliar nitrogen and carbohydrate levels in either tree species, and increased synthesis of condensed tannins and fiber in aspen, but not birch. Elevated O3 decreased nitrogen levels in both tree species and increased production of sugar, condensed tannins, fiber, and lignin in aspen, but not birch. The magnitude of responses to elevated CO2 and O3 varied seasonally for both tree species. When co-occurring, CO2 offset most of the changes in foliar chemistry expressed under elevated O3 alone. Our results suggest that levels of CO2 and O3 predicted for the mid-twenty-first century will alter the foliar chemistry of northern temperate forests with likely consequences for forest community and ecosystem dynamics.

Influence of Genotype, Environment, and Gypsy Moth Herbivory on Local and Systemic Chemical Defenses in Trembling Aspen (Populus tremuloides).

Numerous studies have explored the impacts of intraspecific genetic variation and environment on the induction of plant chemical defenses by herbivory. Relatively few, however, have considered how those factors affect within-plant distribution of induced defenses. This work examined the impacts of plant genotype and soil nutrients on the local and systemic phytochemical responses of trembling aspen (Populus tremuloides) to defoliation by gypsy moth (Lymantria dispar). We deployed larvae onto foliage on individual tree branches for 15 days and then measured chemistry in leaves from: 1) branches receiving damage, 2) undamaged branches of insect-damaged trees, and 3) branches of undamaged control trees. The relationship between post-herbivory phytochemical variation and insect performance also was examined. Plant genotype, soil nutrients, and damage all influenced phytochemistry, with genotype and soil nutrients being stronger determinants than damage. Generally, insect damage decreased foliar nitrogen, increased levels of salicinoids and condensed tannins, but had little effect on levels of a Kunitz trypsin inhibitor, TI3. The largest damage-mediated tannin increases occurred in leaves on branches receiving damage, whereas the largest salicinoid increases occurred in leaves of adjacent, undamaged branches. Foliar nitrogen and the salicinoid tremulacin had the strongest positive and negative relationships, respectively, with insect growth. Overall, plant genetics and environment concomitantly influenced both local and systemic phytochemical responses to herbivory. These findings suggest that herbivory can contribute to phytochemical heterogeneity in aspen foliage, which may in turn influence future patterns of herbivory and nutrient cycling over larger spatial scales.

Herbivore-mediated material fluxes in a northern deciduous forest under elevated carbon dioxide and ozone concentrations.

Anthropogenic changes in atmospheric carbon dioxide (CO2 ) and ozone (O3 ) are known to alter tree physiology and growth, but the cascading effects on herbivore communities and herbivore-mediated nutrient cycling are poorly understood. We sampled herbivore frass, herbivore-mediated greenfall, and leaf-litter deposition in temperate forest stands under elevated CO2 (c. 560 ppm) and O3 (c. 1.5× ambient), analyzed substrate chemical composition, and compared the quality and quantity of fluxes under multiple atmospheric treatments. Leaf-chewing herbivores fluxed 6.2 g m(-2)  yr(-1) of frass and greenfall from the canopy to the forest floor, with a carbon : nitrogen (C : N) ratio 32% lower than that of leaf litter. Herbivore fluxes of dry matter, C, condensed tannins, and N increased under elevated CO2 (35, 32, 63 and 39%, respectively), while fluxes of N decreased (18%) under elevated O3 . Herbivore-mediated dry matter inputs scaled across atmospheric treatments as a constant proportion of leaf-litter inputs. Increased fluxes under elevated CO2 were consistent with increased herbivore consumption and abundance, and with increased plant growth and soil respiration, previously reported for this experimental site. Results suggest that insect herbivory will reinforce other factors, such as photosynthetic rate and fine-root production, impacting C sequestration by forests in future environments.

Plant-associated bacteria degrade defense chemicals and reduce their adverse effects on an insect defoliator.

Phytophagous insects must contend with numerous secondary defense compounds that can adversely affect their growth and development. The gypsy moth (Lymantria dispar) is a polyphagous herbivore that encounters an extensive range of hosts and chemicals. We used this folivore and a primary component of aspen chemical defenses, namely, phenolic glycosides, to investigate if bacteria detoxify phytochemicals and benefit larvae. We conducted insect bioassays using bacteria enriched from environmental samples, analyses of the microbial community in the midguts of bioassay larvae, and in vitro phenolic glycoside metabolism assays. Inoculation with bacteria enhanced larval growth in the presence, but not absence, of phenolic glycosides in the artificial diet. This effect of bacteria on growth was observed only in larvae administered bacteria from aspen foliage. The resulting midgut community composition varied among the bacterial treatments. When phenolic glycosides were included in diet, the composition of midguts in larvae fed aspen bacteria was significantly altered. Phenolic glycosides increased population responses by bacteria that we found able to metabolize these compounds in liquid growth cultures. Several aspects of these results suggest that vectoring or pairwise symbiosis models are inadequate for understanding microbial mediation of plant-herbivore interactions in some systems. First, bacteria that most benefitted larvae were initially foliar residents, suggesting that toxin-degrading abilities of phyllosphere inhabitants indirectly benefit herbivores upon ingestion. Second, assays with single bacteria did not confer the benefits to larvae obtained with consortia, suggesting multi- and inter-microbial interactions are also involved. Our results show that bacteria mediate insect interactions with plant defenses but that these interactions are community specific and highly complex.

Spectroscopic sensitivity of real-time, rapidly induced phytochemical change in response to damage.

An ecological consequence of plant-herbivore interactions is the phytochemical induction of defenses in response to insect damage. Here, we used reflectance spectroscopy to characterize the foliar induction profile of cardenolides in Asclepias syriaca in response to damage, tracked in vivo changes and examined the influence of multiple plant traits on cardenolide concentrations. Foliar cardenolide concentrations were measured at specific time points following damage to capture their induction profile. Partial least-squares regression (PLSR) modeling was employed to calibrate cardenolide concentrations to reflectance spectroscopy. In addition, subsets of plants were either repeatedly sampled to track in vivo changes or modified to reduce latex flow to damaged areas. Cardenolide concentrations and the induction profile of A. syriaca were well predicted using models derived from reflectance spectroscopy, and this held true for repeatedly sampled plants. Correlations between cardenolides and other foliar-related variables were weak or not significant. Plant modification for latex reduction inhibited an induced cardenolide response. Our findings show that reflectance spectroscopy can characterize rapid phytochemical changes in vivo. We used reflectance spectroscopy to identify the mechanisms behind the production of plant secondary metabolites, simultaneously characterizing multiple foliar constituents. In this case, cardenolide induction appears to be largely driven by enhanced latex delivery to leaves following damage.

Rapid phytochemical analysis of birch (Betula) and poplar (Populus) foliage by near-infrared reflectance spectroscopy.

Poplar (Populus) and birch (Betula) species are widely distributed throughout the northern hemisphere, where they are foundation species in forest ecosystems and serve as important sources of pulpwood. The ecology of these species is strongly linked to their foliar chemistry, creating demand for a rapid, inexpensive method to analyze phytochemistry. Our study demonstrates the feasibility of using near-infrared reflectance spectroscopy (NIRS) as an inexpensive, high-throughput tool for determining primary (e.g., nitrogen, sugars, starch) and secondary (e.g., tannins, phenolic glycosides) foliar chemistry of Populus and Betula species, and identifies conditions necessary for obtaining reliable quantitative data. We developed calibrations with high predictive power (residual predictive deviations ≤ 7.4) by relating phytochemical concentrations determined with classical analytical methods (e.g., spectrophotometric assays, liquid chromatography) to NIR spectra, using modified partial least squares regression. We determine that NIRS, although less sensitive and precise than classical methods for some compounds, provides useful predictions in a much faster, less expensive manner than do classical methods.

Atmospheric change alters foliar quality of host trees and performance of two outbreak insect species.

This study examined the independent and interactive effects of elevated carbon dioxide (CO(2)) and ozone (O(3)) on the foliar quality of two deciduous trees species and the performance of two outbreak herbivore species. Trembling aspen (Populus tremuloides) and paper birch (Betula papyrifera) were grown at the Aspen FACE research site in northern Wisconsin, USA, under four combinations of ambient and elevated CO(2) and O(3). We measured the effects of elevated CO(2) and O(3) on aspen and birch phytochemistry and on gypsy moth (Lymantria dispar) and forest tent caterpillar (Malacosoma disstria) performance. Elevated CO(2) nominally affected foliar quality for both tree species. Elevated O(3) negatively affected aspen foliar quality, but only marginally influenced birch foliar quality. Elevated CO(2) slightly improved herbivore performance, while elevated O(3) decreased herbivore performance, and both responses were stronger on aspen than birch. Interestingly, elevated CO(2) largely offset decreased herbivore performance under elevated O(3). Nitrogen, lignin, and C:N were identified as having strong influences on herbivore performance when larvae were fed aspen, but no significant relationships were observed for insects fed birch. Our results support the notion that herbivore performance can be affected by atmospheric change through altered foliar quality, but how herbivores will respond will depend on interactions among CO(2), O(3), and tree species. An emergent finding from this study is that tree age and longevity of exposure to pollutants may influence the effects of elevated CO(2) and O(3) on plant-herbivore interactions, highlighting the need to continue long-term atmospheric change research.