John J. Couture

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.

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 (CO2) and ozone (O3) 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 CO2 and O3. We measured the effects of elevated CO2 and O3 on aspen and birch phytochemistry and on gypsy moth (Lymantria dispar) and forest tent caterpillar (Malacosoma disstria) performance. Elevated CO2 nominally affected foliar quality for both tree species. Elevated O3 negatively affected aspen foliar quality, but only marginally influenced birch foliar quality. Elevated CO2 slightly improved herbivore performance, while elevated O3 decreased herbivore performance, and both responses were stronger on aspen than birch. Interestingly, elevated CO2 largely offset decreased herbivore performance under elevated O3. 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 CO2, O3, 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 CO2 and O3 on plant–herbivore interactions, highlighting the need to continue long-term atmospheric change research.

Increased nitrogen availability influences predator-prey interactions by altering host-plant quality.

Little is known about how plant nutritional and defensive qualities interact to influence predator–prey interactions. To address this need, we provided the neo-tropical milkweed, Asclepias curassavica, with two levels of nitrogen availability and examined how altered host-plant quality influenced the responses of a specialist aphid, Aphis nerii, and a coccinellid predator, Harmonia axyridis. Aphis nerii uses A. curassavica for multiple resources, including nutrition and sequestration of cardenolides for defense against natural enemies. Increased nitrogen availability improved A. curassavica quality by decreasing carbon-to-nitrogen (C:N) ratios and cardenolide concentrations, resulting in A. nerii that also had lower C:N ratios and cardenolide concentrations. Aphis nerii population growth was higher on plants with high nitrogen availability, compared with aphids on plants with low nitrogen availability. In no-choice feeding trials, Harmonia axyridis consumed more high C:N ratio aphids, suggesting a potential compensatory response to reduced aphid nutritional quality. Additionally, H. axyridis were able to consume more low-quality aphids at the expense of increasing exposure to increased cardenolide concentrations, suggesting that interactions between H. axyridis and A. nerii may be strongly influenced by prey nutritional quality. This work highlights the need to consider how variation in plant quality influences herbivore nutritional and defensive quality when examining mechanisms that influence predator–prey interactions.

Long-term exposure to elevated CO2 and O3 alters aspen foliar chemistry across developmental stages

Anthropogenic activities are altering levels of greenhouse gases to the extent that multiple and diverse ecosystem processes are being affected. Two gases that substantially influence forest health are atmospheric carbon dioxide (CO2 ) and tropospheric ozone (O3 ). Plant chemistry will play an important role in regulating ecosystem processes in future environments, but little information exists about the longitudinal effects of elevated CO2 and O3 on phytochemistry, especially for long-lived species such as trees. To address this need, we analysed foliar chemical data from two genotypes of trembling aspen, Populus tremuloides, collected over 10 years of exposure to levels of CO2 and O3 predicted for the year 2050. Elevated CO2 and O3 altered both primary and secondary chemistry, and the magnitude and direction of the responses varied across developmental stages and between aspen genotypes. Our findings suggest that the effects of CO2 and O3 on phytochemical traits that influence forest processes will vary over tree developmental stages, highlighting the need to continue long-term, experimental atmospheric change research.

Insect herbivory alters impact of atmospheric change on northern temperate forests

Stimulation of forest productivity by elevated concentrations of CO2 is expected to partially offset continued increases in anthropogenic CO2 emissions. However, multiple factors can impair the capacity of forests to act as carbon sinks; prominent among these are tropospheric O3 and nutrient limitations1,2. Herbivorous insects also influence carbon and nutrient dynamics in forest ecosystems, yet are often ignored in ecosystem models of forest productivity. Here we assess the effects of elevated levels of CO2 and O3 on insect-mediated canopy damage and organic matter deposition in aspen and birch stands at the Aspen FACE facility in northern Wisconsin, United States. Canopy damage was markedly higher in the elevated CO2 stands, as was the deposition of organic substrates and nitrogen. The opposite trends were apparent in the elevated O3 stands. Using a light-use efficiency model, we show that the negative impacts of herbivorous insects on net primary production more than doubled under elevated concentrations of CO2, but decreased under elevated concentrations of O3. We conclude that herbivorous insects may limit the capacity of forests to function as sinks for anthropogenic carbon emissions in a high CO2 world.

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.

Transgenerational effects of herbivory in a group of long‐lived tree species: maternal damage reduces offspring allocation to resistance traits, but not growth

Summary 1. Numerous studies have explored plant strategies of resource allocation to growth and/or resistance traits within a single generation. In contrast, exceedingly little is known about whether such patterns hold across generations; that is, in seedlings of plants that experienced maternal herbivory. 2. In a common garden study with clonally replicated genotypes of three cottonwood taxa (Populus angustifolia, Populus fremontii and their F1 hybrids), we examined transgenerational response to maternal herbivory in terms of half-sibling seedling offspring (i) germination and growth and (ii) constitutive vs. transgenerational plastic allocation to resistance (measured as both phytochemical content and concentration). Two major results emerged. 3. First, we found that taxa (and often genotypes within a taxon) significantly differed in their constitutive allocation to both growth and resistance. Fremont (P. fremontii) seedlings grew up to seven times more rapidly than did narrowleaf (P. angustifolia) seedlings and had higher or similar content of two key phytochemical resistance traits. Overall, this led to a dilution effect in Fremont relative to narrowleaf, whereby concentrations of two key phytochemical resistance traits were more than 50% lower. 4. Secondly, maternal herbivory by cottonwood leaf beetle larvae on foliage adjacent to developing seeds did not significantly alter offspring growth, but did decrease offspring phytochemical content by 10–55% relative to offspring of maternal control (undamaged) trees. As a result, concentrations of offspring phytochemical resistance traits were reduced by 10–18% in seedlings with maternal herbivory, relative to maternal control seedlings, across all three taxa. These patterns suggest an allocational trade-off, whereby maternal damage results in maintenance of offspring seed size and growth traits at the expense of phytochemical defences in the next generation. 5. Synthesis: This is the first instance in which transgenerational effects of herbivory on growth and defence traits have been described in long-lived, woody plant species. Populus differs substantially from herbaceous plant species or short-lived animals in which transgenerational plasticity of resistance has been examined, in terms of life history (time from germination or hatching to reproductive maturity) and/or in the lag time between generations. These differences may influence the ecological and evolutionary relevance of transgenerational plasticity in defence.

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

AbstractPoplar (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. Graphical abstractNear-infrared reflectance spectroscopy with calibrations based on modified partial least squares regression can provide quantitative measurements of foliar nitrogen, carbohydrate, tannin, and phenolic glycoside content in poplar and birch

Elevated temperature and periodic water stress alter growth and quality of common milkweed ( Asclepias syriaca ) and monarch ( Danaus plexippus ) larval performance

In this study, we examined the independent and interactive effects of temperature and water availability on the growth and foliar traits of common milkweed (Asclepias syriaca) and performance of a specialist herbivore, larvae of the monarch butterfly (Danaus plexippus). Milkweed from multiple population sources collected across a latitudinal gradient in Wisconsin, USA, were grown under all combinations of ambient or elevated temperature and the presence or absence of periodic water stress. Elevated temperature marginally increased, while water stress decreased plant growth. Milkweed from more northerly latitudes experienced larger growth responses to elevated temperature and were more resistant to water stress, especially under higher temperatures. Elevated temperature and water stress also altered milkweed composite foliar trait profiles. Elevated temperature generally increased leaf nitrogen and structural compounds, and decreased leaf mass per area. Water stress also elevated foliar nitrogen, but reduced defensive traits. Monarch larvae performed well on milkweed under elevated temperature and water stress, but gained the most mass on plants exposed to both treatments in combination. Our findings suggest that milkweed populations from more northerly latitudes in the upper Midwest may benefit more from rising temperatures than those in southerly locations, but that these beneficial effects depend on water availability. Monarch larvae grew larger on plants from all experimental treatments relative to ambient condition controls, indicating that future changes in milkweed presence on the landscape will likely influence monarch populations more than the effects of future changes in plant quality on larval performance.

Elevated carbon dioxide and ozone have weak, idiosyncratic effects on herbivorous forest insect abundance, species richness, and community composition

Elevated concentrations of carbon dioxide and tropospheric ozone pose important threats to the abundance, diversity, and composition of forest arthropod communities. In turn, modification of arthropod communities may alter forest health, productivity, and ecosystem services. We studied the independent and interactive effects of elevated CO2 (eCO2) and elevated O3 (eO3) on the abundance, species richness, and community composition of herbivorous arthropods in stands of trembling aspen and paper birch at the Aspen Free Air CO2 Enrichment (FACE) site in northern Wisconsin, USA. We conducted timed, visual surveys of canopy arthropods during each of the summers of 2005, 2006, and 2007. We examined 26 983 arthropods on aspen and 8344 arthropods on birch across the fumigation treatments. Elevated CO2 and eO3 had species‐specific and temporally variable (i.e. idiosyncratic) effects on aspen and birch arthropod abundance and species richness. Weak, idiosyncratic effects of eCO2 and eO3 on herbivorous arthropod abundance and species richness did not significantly alter aspen arthropod community composition but occasionally altered birch insect community composition. Few interactive effects of CO2 and O3 were observed. Growing evidence suggests that the effects of eCO2 and eO3 on communities of insects are difficult to predict because responses are generally weak and species‐ and time‐specific. Although studies to date suggest that impacts of future atmospheres on insect community metrics are likely to be minimal, the possibility remains that effects on particularly important or susceptible species may cascade to alter trophic interactions and, ultimately, ecosystem processes.