New uses for ancient middens: bridging ecological and evolutionary perspectives.

Rodent middens provide a fine-scale spatiotemporal record of plant and animal communities over the late Quaternary. In the Americas, middens have offered insight into biotic responses to past environmental changes and historical factors influencing the distribution and diversity of species. However, few studies have used middens to investigate genetic or ecosystem level responses. Integrating midden studies with neoecology and experimental evolution can help address these gaps and test mechanisms underlying eco-evolutionary patterns across biological and spatiotemporal scales. Fully realizing the potential of middens to answer cross-cutting ecological and evolutionary questions and inform conservation goals in the Anthropocene will require a collaborative research community to exploit existing midden archives and mount new campaigns to leverage midden records globally.

Nutrient conditions mediate mycorrhizal effects on biomass production and cell wall chemistry in poplar.

Large-scale biofuel production from lignocellulosic feedstock is limited by the financial and environmental costs associated with growing and processing lignocellulosic material and the resilience of these plants to environmental stress. Symbiotic associations with arbuscular (AM) and ectomycorrhizal (EM) fungi represent a potential strategy for expanding feedstock production while reducing nutrient inputs. Comparing AM and EM effects on wood production and chemical composition is a necessary step in developing biofuel feedstocks. Here, we assessed the productivity, biomass allocation and secondary cell wall (SCW) composition of greenhouse-grown Populus tremuloidesMichx. inoculated with either AM or EM fungi. Given the long-term goal of reducing nutrient inputs for biofuel production, we further tested the effects of nutrient availability and nitrogen:phosphorus stoichiometry on mycorrhizal responses. Associations with both AM and EM fungi increased plant biomass by 14-74% depending on the nutrient conditions but had minimal effects on SCW composition. Mycorrhizal plants, especially those inoculated with EM fungi, also allocated a greater portion of their biomass to roots, which could be beneficial in the field where plants are likely to experience both water and nutrient stress. Leaf nutrient content was weakly but positively correlated with wood production in mycorrhizal plants. Surprisingly, phosphorus played a larger role in EM plants compared with AM plants. Relative nitrogen and phosphorus availability were correlated with shifts in SCW composition. For AM associations, the benefit of increased wood biomass may be partially offset by increased lignin content, a trait that affects downstream processing of lignocellulosic tissue for biofuels. By comparing AM and EM effects on the productivity and chemical composition of lignocellulosic tissue, this work links broad functional diversity in mycorrhizal associations to key biofuel traits and highlights the importance of considering both biotic and abiotic factors when developing strategies for sustainable biofuel production.

Feast or famine: How is global change affecting forage supply for Yellowstone's ungulate herds?

The ecological integrity of US national parks and other protected areas are under threat in the Anthropocene. For Yellowstone National Park (YNP), the impacts that global change has already had on the park's capacity to sustain its large migratory herds of wild ungulates is incompletely understood. Here we examine how two understudied components of global change, the historical increase in atmospheric CO2 and the spread of nonnative, invasive plant species, may have altered the capacity of YNP to provide forage for ungulates over the last 200-plus years. We performed two experiments: (1) a growth chamber study that determined the growth rates of important invasive and native YNP grasses that are forages for ungulates under preindustrial (280 ppm) versus modern (410 ppm) CO2 levels and (2) a field study that compared the effect of defoliation (clipping) on the shoot growth of invasive and native mesic grassland plants under ambient CO2 conditions in 2019. The growth chamber experiment revealed that modern CO2 increased the growth rates of both invasive and native grasses, and invasive grasses grew faster regardless of CO2 conditions. The field results showed a continuum of positive to negative responses of shoot growth to defoliation, with a subgroup of invasive species responding most positively. Altogether the results indicated that the historical increase in CO2 and the spread of invasive species, some of which were planted to provide forage for ungulates in the early and mid-1900s, have likely increased the capacity of forage production in YNP. However, rising CO2 has also resulted in regional warming and increased aridity in YNP, which will likely reduce grassland productivity. The challenge for global change biologists and park managers is to determine how competing components of global change have already affected and will increasingly affect forage dynamics and the sustainability of Yellowstone's iconic ungulate herds in the Anthropocene.

CO2 studies remain key to understanding a future world.

Contents 34 I. 34 II. 36 III. 37 IV. 37 V. 38 38 References 38 SUMMARY: Characterizing plant responses to past, present and future changes in atmospheric carbon dioxide concentration ([CO2 ]) is critical for understanding and predicting the consequences of global change over evolutionary and ecological timescales. Previous CO2 studies have provided great insights into the effects of rising [CO2 ] on leaf-level gas exchange, carbohydrate dynamics and plant growth. However, scaling CO2 effects across biological levels, especially in field settings, has proved challenging. Moreover, many questions remain about the fundamental molecular mechanisms driving plant responses to [CO2 ] and other global change factors. Here we discuss three examples of topics in which significant questions in CO2 research remain unresolved: (1) mechanisms of CO2 effects on plant developmental transitions; (2) implications of rising [CO2 ] for integrated plant-water dynamics and drought tolerance; and (3) CO2 effects on symbiotic interactions and eco-evolutionary feedbacks. Addressing these and other key questions in CO2 research will require collaborations across scientific disciplines and new approaches that link molecular mechanisms to complex physiological and ecological interactions across spatiotemporal scales.

Host Plant Physiology and Mycorrhizal Functioning Shift across a Glacial through Future [CO2] Gradient.

Rising atmospheric carbon dioxide concentration ([CO2]) may modulate the functioning of mycorrhizal associations by altering the relative degree of nutrient and carbohydrate limitations in plants. To test this, we grew Taraxacum ceratophorum and Taraxacum officinale (native and exotic dandelions) with and without mycorrhizal fungi across a broad [CO2] gradient (180-1,000 µL L-1). Differential plant growth rates and vegetative plasticity were hypothesized to drive species-specific responses to [CO2] and arbuscular mycorrhizal fungi. To evaluate [CO2] effects on mycorrhizal functioning, we calculated response ratios based on the relative biomass of mycorrhizal (MBio) and nonmycorrhizal (NMBio) plants (RBio = [MBio - NMBio]/NMBio). We then assessed linkages between RBio and host physiology, fungal growth, and biomass allocation using structural equation modeling. For T. officinale, RBio increased with rising [CO2], shifting from negative to positive values at 700 µL L-1 [CO2] and mycorrhizal effects on photosynthesis and leaf growth rates drove shifts in RBio in this species. For T. ceratophorum, RBio increased from 180 to 390 µL L-1 and further increases in [CO2] caused RBio to shift from positive to negative values. [CO2] and fungal effects on plant growth and carbon sink strength were correlated with shifts in RBio in this species. Overall, we show that rising [CO2] significantly altered the functioning of mycorrhizal associations. These symbioses became more beneficial with rising [CO2], but nonlinear effects may limit plant responses to mycorrhizal fungi under future [CO2]. The magnitude and mechanisms driving mycorrhizal-CO2 responses reflected species-specific differences in growth rate and vegetative plasticity, indicating that these traits may provide a framework for predicting mycorrhizal responses to global change.

A dynamic leaf gas-exchange strategy is conserved in woody plants under changing ambient CO2 : evidence from carbon isotope discrimination in paleo and CO2 enrichment studies.

Rising atmospheric [CO2 ], ca , is expected to affect stomatal regulation of leaf gas-exchange of woody plants, thus influencing energy fluxes as well as carbon (C), water, and nutrient cycling of forests. Researchers have proposed various strategies for stomatal regulation of leaf gas-exchange that include maintaining a constant leaf internal [CO2 ], ci , a constant drawdown in CO2 (ca  - ci ), and a constant ci /ca . These strategies can result in drastically different consequences for leaf gas-exchange. The accuracy of Earth systems models depends in part on assumptions about generalizable patterns in leaf gas-exchange responses to varying ca . The concept of optimal stomatal behavior, exemplified by woody plants shifting along a continuum of these strategies, provides a unifying framework for understanding leaf gas-exchange responses to ca . To assess leaf gas-exchange regulation strategies, we analyzed patterns in ci inferred from studies reporting C stable isotope ratios (δ(13) C) or photosynthetic discrimination (∆) in woody angiosperms and gymnosperms that grew across a range of ca spanning at least 100 ppm. Our results suggest that much of the ca -induced changes in ci /ca occurred across ca spanning 200 to 400 ppm. These patterns imply that ca  - ci will eventually approach a constant level at high ca because assimilation rates will reach a maximum and stomatal conductance of each species should be constrained to some minimum level. These analyses are not consistent with canalization toward any single strategy, particularly maintaining a constant ci . Rather, the results are consistent with the existence of a broadly conserved pattern of stomatal optimization in woody angiosperms and gymnosperms. This results in trees being profligate water users at low ca , when additional water loss is small for each unit of C gain, and increasingly water-conservative at high ca , when photosystems are saturated and water loss is large for each unit C gain.

Evolutionary history underlies plant physiological responses to global change since the last glacial maximum.

Assessing family- and species-level variation in physiological responses to global change across geologic time is critical for understanding factors that underlie changes in species distributions and community composition. Here, we used stable carbon isotopes, leaf nitrogen content and stomatal measurements to assess changes in leaf-level physiology in a mixed conifer community that underwent significant changes in composition since the last glacial maximum (LGM) (21 kyr BP). Our results indicate that most plant taxa decreased stomatal conductance and/or maximum photosynthetic capacity in response to changing conditions since the LGM. However, plant families and species differed in the timing and magnitude of these physiological responses, and responses were more similar within families than within co-occurring species assemblages. This suggests that adaptation at the level of leaf physiology may not be the main determinant of shifts in community composition, and that plant evolutionary history may drive physiological adaptation to global change over recent geologic time.

Host identity impacts rhizosphere fungal communities associated with three alpine plant species.

Fungal diversity and composition are still relatively unknown in many ecosystems; however, host identity and environmental conditions are hypothesized to influence fungal community assembly. To test these hypotheses, we characterized the richness, diversity, and composition of rhizosphere fungi colonizing three alpine plant species, Taraxacum ceratophorum, Taraxacum officinale, and Polemonium viscosum. Roots were collected from open meadow and willow understory habitats at treeline on Pennsylvania Mountain, Colorado, USA. Fungal small subunit ribosomal DNA was sequenced using fungal-specific primers, sample-specific DNA tags, and 454 pyrosequencing. We classified operational taxonomic units (OTUs) as arbuscular mycorrhizal (AMF) or non-arbuscular mycorrhizal (non-AMF) fungi and then tested whether habitat or host identity influenced these fungal communities. Approximately 14% of the sequences represented AMF taxa (44 OTUs) with the majority belonging to Glomus groups A and B. Non-AMF sequences represented 186 OTUs belonging to Ascomycota (58%), Basidiomycota (26%), Zygomycota (14%), and Chytridiomycota (2%) phyla. Total AMF and non-AMF richness were similar between habitats but varied among host species. AMF richness and diversity per root sample also varied among host species and were highest in T. ceratophorum compared with T. officinale and P. viscosum. In contrast, non-AMF richness and diversity per root sample were similar among host species except in the willow understory where diversity was reduced in T. officinale. Fungal community composition was influenced by host identity but not habitat. Specifically, T. officinale hosted a different AMF community than T. ceratophorum and P. viscosum while P. viscosum hosted a different non-AMF community than T. ceratophorum and T. officinale. Our results suggest that host identity has a stronger effect on rhizosphere fungi than habitat. Furthermore, although host identity influenced both AMF and non-AMF, this effect was stronger for the mutualistic AMF community.

Soil fungal effects on floral signals, rewards, and aboveground interactions in an alpine pollination web.

PREMISE OF THE STUDY: Plants interact with above- and belowground organisms; the combined effects of these interactions determine plant fitness and trait evolution. To better understand the ecological and evolutionary implications of multispecies interactions, we explored linkages between soil fungi, pollinators, and floral larcenists in Polemonium viscosum (Polemoniaceae). METHODS: Using a fungicide, we experimentally reduced fungal colonization of krummholz and tundra P. viscosum in 2008-2009. We monitored floral signals and rewards, interactions with pollinators and larcenists, and seed set for fungicide-treated and control plants. KEY RESULTS: Fungicide effects varied among traits, between interactions, and with environmental context. Treatment effects were negligible in 2008, but stronger in 2009, especially in the less-fertile krummholz habitat. There, fungicide increased nectar sugar content and damage by larcenist ants, but did not affect pollination. Surprisingly, fungicide also enhanced seed set, suggesting that direct resource costs of soil fungi exceed indirect benefits from reduced larceny. In the tundra, fungicide effects were negligible in both years. However, pooled across treatments, colonization by mycorrhizal fungi in 2009 correlated negatively with the intensity and diversity of floral volatile organic compounds, suggesting integrated above- and belowground signaling pathways. CONCLUSIONS: Fungicide effects on floral rewards in P. viscosum link soil fungi to ecological costs of pollinator attraction. Trait-specific linkages to soil fungi should decouple expression of sensitive and buffered floral phenotypes in P. viscosum. Overall, this study demonstrates how multitrophic linkages may lead to shifting selection pressures on interaction traits, restricting the evolution of specialization.