Genotype, mycorrhizae, and herbivory interact to shape strawberry plant functional traits.

Arbuscular mycorrhizal fungi (AMF) and herbivores are ubiquitous biotic agents affecting plant fitness. While individual effects of pairwise interactions have been well-studied, less is known about how species interactions above and belowground interact to influence phenotypic plasticity in plant functional traits, especially phytochemicals. We hypothesized that mycorrhizae would mitigate negative herbivore effects by enhancing plant physiology and reproductive traits. Furthermore, we expected genotypic variation would influence functional trait responses to these biotic agents. To test these hypotheses, we conducted a manipulative field-based experiment with three strawberry (Fragaria x ananassa) genotypes to evaluate plant phenotypic plasticity in multiple functional traits. We used a fully-crossed factorial design in which plants from each genotype were exposed to mycorrhizal inoculation, herbivory, and the combined factors to examine effects on plant growth, reproduction, and floral volatile organic compounds (VOCs). Genotype and herbivory were key determinants of phenotypic variation, especially for plant physiology, biomass allocation, and floral volatiles. Mycorrhizal inoculation increased total leaf area, but only in plants that received no herbivory, and also enhanced flower and fruit numbers across genotypes and herbivory treatments. Total fruit biomass increased for one genotype, with up to 30-40% higher overall yield depending on herbivory. Herbivory altered floral volatile profiles and increased total terpenoid emissions. The effects of biotic treatments, however, were less important than the overall influence of genotype on floral volatile composition and emissions. This study demonstrates how genotypic variation affects plant phenotypic plasticity to herbivory and mycorrhizae, playing a key role in shaping physiological and phytochemical traits that directly and indirectly influence productivity.

Genotypic variation in floral volatiles influences floral microbiome more strongly than interactions with herbivores and mycorrhizae in strawberriesd.

The floral microbiome is of significant relevance to plant reproduction and crop productivity. While plant genotype is key to floral microbiome assembly, whether and how genotypic variation in floral traits and plant-level mutualistic and antagonistic interactions at the rhizosphere and phyllosphere influence the microbiome in the anthosphere remain little known. Using a factorial field experiment that manipulated biotic interactions belowground (mycorrhizae treatments) and aboveground (herbivory treatments) in three strawberry genotypes, we assessed how genotypic variation in flower abundance and size and plant-level biotic interactions influence the bidirectional relationships between floral volatile organic compounds (VOCs) and the floral microbiome using structural equation modeling. We found that plant genotype played a stronger role, overall, in shaping the floral microbiome than biotic interactions with mycorrhizae and herbivores. Genotypic variation in flower abundance and size influenced the emission of floral VOCs, especially terpenes (e.g. α- and ß-pinene, ocimene isomers) and benzenoids (e.g. p-anisaldehyde, benzaldehyde), which in turn affected floral bacterial and fungal communities. While the effects of biotic interactions on floral traits including VOCs were weak, mycorrhizae treatments (mycorrhizae and herbivory + mycorrhizae) affected the fungal community composition in flowers. These findings improve our understanding of the mechanisms by which plant genotype influences floral microbiome assembly and provide the first evidence that biotic interactions in the rhizosphere and phyllosphere can influence the floral microbiome, and offer important insights into agricultural microbiomes.

Root Secondary Metabolites in Populus tremuloides: Effects of Simulated Climate Warming, Defoliation, and Genotype.

Climate warming can influence interactions between plants and associated organisms by altering levels of plant secondary metabolites. In contrast to studies of elevated temperature on aboveground phytochemistry, the consequences of warming on root chemistry have received little attention. Herein, we investigated the effects of elevated temperature, defoliation, and genotype on root biomass and phenolic compounds in trembling aspen (Populus tremuloides). We grew saplings of three aspen genotypes under ambient or elevated temperatures (+4-6 °C), and defoliated (by 75%) half of the trees in each treatment. After 4 months, we harvested roots and determined their condensed tannin and salicinoid (phenolic glycoside) concentrations. Defoliation reduced root biomass, with a slightly larger impact under elevated, relative to ambient, temperature. Elevated temperature decreased condensed tannin concentrations by 21-43% across the various treatment combinations. Warming alone did not alter salicinoid concentrations but eliminated a small negative impact of defoliation on those compounds. Graphical vector analysis suggests that effects of warming and defoliation on condensed tannins and salicinoids were predominantly due to reduced biosynthesis of these metabolites in roots, rather than to changes in root biomass. In general, genotypes did not differ in their responses to temperature or temperature by defoliation interactions. Collectively, our results suggest that future climate warming will alter root phytochemistry, and that effects will vary among different classes of secondary metabolites and be influenced by concurrent ecological interactions such as herbivory. Temperature- and herbivory-mediated changes in root chemistry have the potential to influence belowground trophic interactions and soil nutrient dynamics.

The effects of urbanization on bee communities depends on floral resource availability and bee functional traits.

Wild bees are important pollinators in many ecosystems threatened by anthropogenic disturbance. Urban development can reduce and degrade natural habitat for bees and other pollinators. However, some researchers suggest that cities could also provide refuge for bees, given that agricultural intensification may pose a greater risk. In this study, we surveyed bee communities at 15 farms and gardens across an urban-rural gradient in southeastern Michigan, USA to evaluate the effect of urbanization on bees. We examined how floral resources, bee functional traits, temperature, farm size, and the spatial scale of analysis influence bee response to urbanization. We found that urbanization positively affected bee diversity and evenness but had no effect on total abundance or species richness. Additionally, urbanization altered bee community composition via differential effects on bee species and functional groups. More urbanized sites supported a greater number of exotic, above-ground nesting, and solitary bees, but fewer eusocial bees. Blooming plant species richness positively influenced bee species diversity and richness. Furthermore, the amount of available floral resources was positively associated with exotic and eusocial bee abundances. Across sites, nearly 70% of floral resources were provided by exotic plants, most of which are characterized as weedy but not invasive. Our study demonstrates that urbanization can benefit some bee species and negatively impact others. Notably, Bombus and Lasioglossum (Dialictus), were two important pollinator groups negatively affected by urbanization. Our study supports the idea that urban environments can provide valuable habitat for diverse bee communities, but demonstrates that some bees are vulnerable to urbanization. Finally, while our results indicate that increasing the abundance and richness of floral resources could partially compensate for negative effects of urbanization on bees, the effectiveness of such measures may be limited by other factors, such as urban warming.

Does urbanization favour exotic bee species? Implications for the conservation of native bees in cities.

A growing body of research indicates that cities can support diverse bee communities. However, urbanization may disproportionately benefit exotic bees, potentially to the detriment of native species. We examined the influence of urbanization on exotic and native bees using two datasets from Michigan, USA. We found that urbanization positively influenced exotic-but not native-bee abundance and richness, and that this association could not be explained by proximity to international ports of entry, prevalence of exotic flora or urban warming. We found a negative relationship between native and exotic bee abundance at sites with high total bee abundance, suggesting that exotic bees may negatively affect native bee populations. These effects were not driven by the numerically dominant exotic honeybee, but rather by other exotic bees. Our findings complicate the emerging paradigm of cities as key sites for pollinator conservation.

Global change effects on plant-insect interactions: the role of phytochemistry.

Natural and managed ecosystems are undergoing rapid environmental change due to a growing human population and associated increases in industrial and agricultural activity. Global environmental change directly and indirectly impacts insect herbivores and pollinators. In this review, we highlight recent research examining how environmental change factors affect plant chemistry and, in turn, ecological interactions among plants, herbivores, and pollinators. Recent studies reveal the complex nature of understanding global change effects on plant secondary metabolites and plant-insect interactions. Nonetheless, these studies indicate that phytochemistry mediates insect responses to environmental change. Future research on the chemical ecology of plant-insect interactions will provide critical insight into the ecological effects of climate change and other anthropogenic disturbances. We recommend greater attention to investigations examining interactive effects of multiple environmental change factors in addition to chemically mediated plant-pollinator interactions, given limited research in these areas.

Experimental climate warming alters aspen and birch phytochemistry and performance traits for an outbreak insect herbivore.

Climate change and insect outbreaks are key factors contributing to regional and global patterns of increased tree mortality. While links between these environmental stressors have been established, our understanding of the mechanisms by which elevated temperature may affect tree-insect interactions is limited. Using a forest warming mesocosm, we investigated the influence of elevated temperature on phytochemistry, tree resistance traits, and insect performance. Specifically, we examined warming effects on forest tent caterpillar (Malacosoma disstria) and host trees aspen (Populus tremuloides) and birch (Betula papyrifera). Trees were grown under one of three temperature treatments (ambient, +1.7 °C, +3.4 °C) in a multiyear open-air warming experiment. In the third and fourth years of warming (2011, 2012), we assessed foliar nutrients and defense chemistry. Elevated temperatures altered foliar nitrogen, carbohydrates, lignin, and condensed tannins, with differences in responses between species and years. In 2012, we performed bioassays using a common environment approach to evaluate plant-mediated indirect warming effects on larval performance. Warming resulted in decreased food conversion efficiency and increased consumption, ultimately with minimal effect on larval development and biomass. These changes suggest that insects exhibited compensatory feeding due to reduced host quality. Within the context of observed phytochemical variation, primary metabolites were stronger predictors of insect performance than secondary metabolites. Between-year differences in phytochemical shifts corresponded with substantially different weather conditions during these two years. By sampling across years within an ecologically realistic and environmentally open setting, our study demonstrates that plant and insect responses to warming can be temporally variable and context dependent. Results indicate that elevated temperatures can alter phytochemistry, tree resistance traits, and herbivore feeding, but that annual weather variability may modulate warming effects leading to uncertain consequences for plant-insect interactions with projected climate change.

Simulated climate warming alters phenological synchrony between an outbreak insect herbivore and host trees.

As the world's climate warms, the phenologies of interacting organisms in seasonally cold environments may advance at differing rates, leading to alterations in phenological synchrony that can have important ecological consequences. For temperate and boreal species, the timing of early spring development plays a key role in plant-herbivore interactions and can influence insect performance, outbreak dynamics, and plant damage. We used a field-based, meso-scale free-air forest warming experiment (B4WarmED) to examine the effects of elevated temperature on the phenology and performance of forest tent caterpillar (Malacosoma disstria) in relation to the phenology of two host trees, aspen (Populus tremuloides) and birch (Betula papyrifera). Results of our 2-year study demonstrated that spring phenology advanced for both insects and trees, with experimentally manipulated increases in temperature of 1.7 and 3.4 °C. However, tree phenology advanced more than insect phenology, resulting in altered phenological synchrony. Specifically, we observed a decrease in the time interval between herbivore egg hatch and budbreak of aspen in both years and birch in one year. Moreover, warming decreased larval development time from egg hatch to pupation, but did not affect pupal mass. Larvae developed more quickly on aspen than birch, but pupal mass was not affected by host species. Our study reveals that warming-induced phenological shifts can alter the timing of ecological interactions across trophic levels. These findings illustrate one mechanism by which climate warming could mediate insect herbivore outbreaks, and also highlights the importance of climate change effects on trophic interactions.

Interactive effects of simulated nitrogen deposition and altered precipitation patterns on plant allelochemical concentrations.

Global environmental change alters the supply of multiple limiting resources that regulate plant primary and secondary metabolism. Through modifications in resource availability, acquisition, and allocation, global change is likely to influence plant chemical defenses, and consequently species interactions that are mediated by these compounds. While many studies focus on individual global change factors, simultaneous changes in abiotic factors may interact to influence plant allelochemicals. In this study, we examined the individual and interactive effects of nitrogen enrichment and altered precipitation patterns on chemical defense compounds (iridoid glycosides) of an invasive plant, Linaria dalmatica. Plants were grown from seed in native mixed-grass prairie for 2 years. Nitrogen and water treatments were applied in each growing season over this period. Results indicate that soil water and nitrogen availability interact to shape plant chemical defense concentrations in L. dalmatica. Nitrogen addition decreased iridoid glycoside concentrations by approximately 25% under reduced water availability, increased concentrations by 37% in ambient water plots, and had no effect on these chemical defenses for plants growing under augmented water supply. Thus, results show differing patterns of allelochemical response to nitrogen enrichment, with respect to both the magnitude and direction of change, depending on water availability. Our study demonstrates the importance of examining multiple environmental factors in order to predict potential changes in plant chemical defenses with climate change.

Nitrogen enrichment differentially affects above- and belowground plant defense.

PREMISE OF THE STUDY: Human nitrogen (N) inputs to terrestrial ecosystems have greatly increased in recent years and may have important consequences for plant growth, reproduction, and defense. Although numerous studies have investigated the effects of nitrogen addition on plants, few have examined both above- and belowground responses within a range of predicted increase and apart from concomitant increases in other nutrients. • METHODS: We conducted a greenhouse experiment to study the consequences of increased nitrogen inputs, such as those from atmospheric N deposition, on plant performance, chemical defenses, and allocation tradeoffs for an invasive species, Linaria dalmatica. This plant produces iridoid glycosides, which are a group of terpenoid compounds. • KEY RESULTS: Soil nitrogen enrichment increased growth, reproduction, and whole-plant iridoid glycosides while decreasing some costs of defense. Interestingly, nitrogen addition had varying effects on defense allocation to above- and belowground tissues. Specifically, there was no change in iridoid glycoside concentrations of shoots, whereas concentrations decreased in flowers by ~35% and increased in roots by >400%. • CONCLUSIONS: Observed increases in plant performance and chemical defenses may have implications for the invasion potential of L. dalmatica. Moreover, our results highlight the importance of evaluating both above- and belowground plant defenses. In particular, findings presented here indicate that research focused on leaf-level defenses may not detect key allelochemical responses, including changes in plant resistance traits that could affect consumers (e.g., herbivores and pathogens) that specialize on different plant tissues as well as plant fitness and invasion success.

Iridoid glycoside variation in the invasive plant Dalmatian toadflax, Linaria dalmatica (Plantaginaceae), and sequestration by the biological control agent, Calophasia lunula.

Invasive plant species can have significant ecological and economic impacts. Although numerous hypotheses highlight the importance of the chemical defenses of invasive plant species, the chemical ecology of many invasive plants has not yet been investigated. In this study, we provide the first quantitative investigation of variation in iridoid glycoside concentrations of the invasive plant Dalmatian toadflax (Linaria dalmatica). We examined variation in chemical defenses at three levels: (1) variation within and among populations; (2) variation due to phenology and/or seasonal differences; and (3) variation among plant parts (leaves, flowers, and stems). Further, we examined two biological control agents introduced to control L. dalmatica for the ability to sequester iridoid glycosides from this invasive plant. Results indicate that L. dalmatica plants can contain high concentrations of iridoid glycosides (up to 17.4% dry weight of leaves; mean = 6.28 ± 0.5 SE). We found significant variation in iridoid glycoside concentrations both within and among plant populations, over the course of the growing season, and among plant parts. We also found that one biological control agent, Calophasia lunula (Lepidoptera: Noctuidae), was capable of sequestering antirrhinoside, an iridoid glycoside found in L. dalmatica, at levels ranging from 2.7 to 7.5% dry weight. A second biological control agent, Mecinus janthinus (Coleoptera: Curculionidae), a stem-mining weevil, did not sequester iridoid glycosides. The demonstrated variation in L. dalmatica chemical defenses may have implications for understanding variation in the degree of invasiveness of different populations as well as variation in the efficacy of biological control efforts.

Underutilized resources for studying the evolution of invasive species during their introduction, establishment, and lag phases.

The early phases of biological invasions are poorly understood. In particular, during the introduction, establishment, and possible lag phases, it is unclear to what extent evolution must take place for an introduced species to transition from established to expanding. In this study, we highlight three disparate data sources that can provide insights into evolutionary processes associated with invasion success: biological control organisms, horticultural introductions, and natural history collections. All three data sources potentially provide introduction dates, information about source populations, and genetic and morphological samples at different time points along the invasion trajectory that can be used to investigate preadaptation and evolution during the invasion process, including immediately after introduction and before invasive expansion. For all three data sources, we explore where the data are held, their quality, and their accessibility. We argue that these sources could find widespread use with a few additional pieces of data, such as voucher specimens collected at certain critical time points during biocontrol agent quarantine, rearing, and release and also for horticultural imports, neither of which are currently done consistently. In addition, public access to collected information must become available on centralized databases to increase its utility in ecological and evolutionary research.