Ken Keefover-Ring

Resources play an important role in shaping the evolution of plant defensive strategies. How resource availability influences overall phytochemical diversity within species, and how this in‐turn affects herbivore damage is not well understood. Using Monarda fistulosa, a species that produces diverse terpene compounds and distinct chemotypes (i.e. thymol or carvacrol), we addressed three questions: 1) how do phytochemical components (i.e. total concentration, Shannon diversity, and chemotypes) vary between distinct populations from two large, widely separated regions that differ in climate and productivity? 2) How do these components influence attack by different herbivore species? 3) What are the growth costs of higher phytochemical levels in different chemotypes? Seeds were collected from 12 populations spread across two regions that differ in precipitation‐driven productivity, six from a low‐productivity environment in Montana and six from a high‐productivity environment in Wisconsin, and grown in a common garden in Wisconsin. We analyzed terpene concentrations, quantified the number of herbivores and damage, and measured plant above‐ground biomass. Plants from Montana exhibited higher terpene concentrations but lower diversity, whereas those from Wisconsin displayed lower terpene concentrations but higher diversity. Plants from Wisconsin received more damage from a specialist seed predator and leaf‐chewing herbivores, although abundance of a specialist aphid was lower, compared to plants from Montana. Total terpene concentration emerged as the primary predictor of herbivore damage, although some differences were observed between region of origin and with terpene richness. Herbivores exhibited mixed responses to phytochemistry; some herbivores were negatively affected, while a specialized leaf galler responded positively. Costs of producing defenses were evident by negative correlations between plant biomass and total terpene concentration (but not chemical richness or Shannon diversity). Our study revealed that regional differences in productivity can influence the evolution of phytochemical concentration and diversity, which, in turn, affects plant damage and incurs growth‐related costs.

Coevolution between plants and their animal predators has led to diverse defensive adaptations. Multiple theories of defense propose that there are resource allocation costs associated with producing chemical defenses. One leading hypothesis, optimal defense theory (ODT), suggests that natural selection will result in the allocation of resources to defenses that optimize the cost‐to‐benefit ratio between defense and other functional processes. The population decline of the rare subalpine wetland species, Arizona willow ( Salix arizonica ), has been attributed to various biotic and abiotic factors, with browsing from wild and domestic ungulates as a significant concern for at least three decades. In a field experiment using natural populations, we compare the relationship between phytochemical defense and height in Arizona willows with and without long‐term protection from browsing via browse exclosures. Consistent with the predictions of ODT, individuals with physical protection from ungulate browsing for multiple years had significantly lower phenolic glycoside (PG) concentrations and increased plant height compared to unprotected individuals. A similar pattern was found across all individuals, whereby total PG concentration and height were negatively correlated. In a short‐term experiment in natural populations, changes in levels of defense were not observed when plants received protection for only one growing season. The contrasting pattern of defense plasticity in response to long‐term versus short‐term physical protection suggests a differential plastic response in this long‐lived species. Delayed reduction in PG concentration may serve as a benefit to avoid mismatches between environmental cues and responses. Our research sheds light on the intricate dynamics between plant‐defense strategies, environmental pressures, and evolutionary adaptations in shaping plant–browser interactions.

The current study aimed to evaluate the presence of chemical variations in essential oils (EOs) extracted from Artemisia scoparia growing at different altitudes and to reveal their antibacterial, mosquito larvicidal, and repellent activity. The gas chromatographic–mass spectrometric analysis of A. scoparia EOs revealed that the major compounds were capillene (9.6–31.8%), methyleugenol (0.2–26.6%), β-myrcene (1.9–21.4%), γ-terpinene (1.5–19.4%), trans-β-caryophyllene (0.8–12.4%), and eugenol (0.1–9.1%). The EO of A. scoparia collected from the city of Attock at low elevation was the most active against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa bacteria (minimum inhibitory concentration of 156–1250 µg/mL) and showed the best mosquito larvicidal activity (LC₅₀, 55.3 mg/L). The EOs of A. scoparia collected from the high-altitude areas of Abbottabad and Swat were the most repellent for females of Ae. aegypti and exhibited repellency for 120 min and 165 min, respectively. The results of the study reveal that different climatic conditions and altitudes have significant effects on the chemical compositions and the biological activity of essential oils extracted from the same species.

Plants produce a wide variety of secondary metabolites, but intraspecific variation in space and time can alter the ecological interactions these compounds mediate. The aim of this work was to document the spatial and temporal chemical biogeography ofMonarda fistulosa. I collected leaves from 1587M. fistulosaindividuals from 86 populations from Colorado to Manitoba, extracted and analyzed their terpenes with gas chromatography, mapped monoterpene chemotypes, and analyzed chemical variation with principal component analysis. I also measured the amounts of terpenes in different plant tissues to examine intraplant variation and monitored leaf terpene chemistry over a single growing season to examine temporal patterns. Finally, I extracted terpenes from herbarium samples up to 125 years old and compared the chemotypes with recent samples from the same sites.M. fistulosapopulations consisted mostly of thymol (T) and carvacrol (C) chemotypes, but geraniol (G) and (R)‐(−)‐linalool (L), a chemotype new to this species, were also present. A principal component analysis showed that most of the chemical variation across populations was due to the amounts of the dominant terpene in plants. Intraplant tissue chemistry revealed that leaves mostly had the greatest amounts of terpenes, followed by floral structures, stems, and roots. Short‐term temporal variation in leaf chemistry of T and C plants over a growing season showed that plants produced the highest levels of biosynthetic precursors early in the season and their dominant monoterpenes peaked in mid‐summer. Plant chemotype was discernable in the oldest herbarium samples, and 15 of 18 historic samples matched the majority chemotype currently at the site, indicating that population chemotype ratios may remain stable over longer time scales. Overall, the results show that plant species' secondary chemistry can vary both spatially and temporally, which may alter the biological interactions that these compounds mediate over space and time.

Secondary chemistry often differs between sexes in dioecious plant species, a pattern attributed to its possible role in the evolution and/or maintenance of dioecy. We used GC-MS to measure floral volatiles emitted from, and LC-MS to quantitate non-volatile secondary compounds contained in, female and male Salix purpurea willow catkins from an F₂ family. Using the abundance of these chemicals, we then performed quantitative trait locus (QTL) mapping to locate them on the genome, identified biosynthetic candidate genes in the QTL intervals, and examined expression patterns of candidate genes using RNA-seq. Male flowers emitted more total terpenoids than females, but females produced more benzenoids. Male tissue contained greater amounts of phenolic glycosides, but females had more chalcones and flavonoids. A flavonoid pigment and a spermidine derivative were found only in males. Male catkins were almost twice the mass of females. Forty-two QTL were mapped for 25 chemical traits and catkin mass across 16 of the 19 S. purpurea chromosomes. Several candidate genes were identified, including a chalcone isomerase associated with seven compounds. A better understanding of the genetic basis of the sexually dimorphic chemistry of a dioecious species may shed light on how chemically mediated ecological interactions may have helped in the evolution and maintenance of dioecy.

Chiropterophily, or bat pollination, is typically considered a highly specialized pollination system that has evolved independently numerous times across the angiosperm phylogeny, with distinct lineages often converging on a similar suite of floral traits. The African baobab,Adansonia digitata, occurs widespread across continental Africa and introduced throughout much of the tropics, possesses floral traits classically associated with bat pollination, namely nocturnal anthesis, pendulous white flowers, and a “musky” fragrance. Despite this, our observations and pollination exclusion experiments in South African baobab populations suggested little if any role for bats, but instead showed that hawkmoths are the main pollinators. Hand pollination indicated strong self‐incompatibility and crossing experiments suggest minimal diurnal receptivity. Furthermore, our analyses of floral volatiles revealed not only sulfur‐containing compounds, commonly associated with bat pollination, but also a high concentration of the sesquiterpene β‐caryophyllene, a compound generally more typical of hawkmoth pollination. Comparing previous pollination studies and published floral scent profiles from West Africa suggests that the classic bat pollination system in baobabs may be more labile than previously thought. This study provides an empirical example of a species that most likely evolved due to bat pollination yet has some degree of generality and possible geographic variation in floral traits and pollinator visitation patterns across its range.

All organisms experience fundamental conflicts between divergent metabolic processes. In plants, a pivotal conflict occurs between allocation to growth, which accelerates resource acquisition, and to defense, which protects existing tissue against herbivory. Trade-offs between growth and defense traits are not universally observed, and a central prediction of plant evolutionary ecology is that context-dependence of these trade-offs contributes to the maintenance of intraspecific variation in defense [Züst and Agrawal, Annu. Rev. Plant Biol., 68, 513–534 (2017)]. This prediction has rarely been tested, however, and the evolutionary consequences of growth–defense trade-offs in different environments are poorly understood, especially in long-lived species [Cipollini et al., Annual Plant Reviews (Wiley, 2014), pp. 263–307]. Here we show that intraspecific trait trade-offs, even when fixed across divergent environments, interact with competition to drive natural selection of tree genotypes corresponding to their growth–defense phenotypes. Our results show that a functional trait trade-off, when coupled with environmental variation, causes real-time divergence in the genetic architecture of tree populations in an experimental setting. Specifically, competitive selection for faster growth resulted in dominance by fast-growing tree genotypes that were poorly defended against natural enemies. This outcome is a signature example of eco-evolutionary dynamics: Competitive interactions affected microevolutionary trajectories on a timescale relevant to subsequent ecological interactions [Brunner et al., Funct. Ecol. 33, 7–12 (2019)]. Eco-evolutionary drivers of tree growth and defense are thus critical to stand-level trait variation, which structures communities and ecosystems over expansive spatiotemporal scales. Significance Genetic diversity is critical for ecological resilience and is maintained when different traits are advantageous in different environments. Traits are not entirely independent, however: When trade-offs occur, selection on one trait may indirectly act on another. Here we examine how environmentally mediated selection for growth interacts with a trade-off between growth and herbivore defense in experimental aspen stands. We show that even an environmentally fixed trait trade-off can lead to real-time divergence in population genetic composition between environments. The underlying growth–defense trade-off led to an eco-evolutionary dynamic wherein high-competition environments selected for fast-growing trees and thus ultimately selected against herbivore resistance. This discovery reveals how patterns of genetic covariation, coupled with environmental variation, can link ecological and evolutionary processes.

The development of non-recombining sex chromosomes has radical effects on the evolution of discrete sexes and sexual dimorphism. Although dioecy is rare in plants, sex chromosomes have evolved repeatedly throughout the diversification of angiosperms, and many of these sex chromosomes are relatively young compared to those found in vertebrates. In this study, we designed and used a sequence capture array to identify a novel sex-linked region (SLR) in Salix nigra, a basal species in the willow clade, and demonstrated that this species has XY heterogamety. We did not detect any genetic overlap with the previously characterized ZW SLRs in willows, which map to a different chromosome. The S. nigra SLR is characterized by strong recombination suppression across a 2 MB region and an excess of low-frequency alleles, resulting in a low Tajima’s D compared to the remainder of the genome. We speculate that either a recent bottleneck in population size or factors related to positive or background selection generated this differential pattern of Tajima’s D on the X and autosomes. This discovery provides insights into factors that may influence the evolution of sex chromosomes in plants and contributes to a large number of recent observations that underscore their dynamic nature.

Summary: The ability to tolerate neighboring plants (i.e. degree of competitive response) is a key determinant of plant success in high‐competition environments. Plant genotypes adjust their functional trait expression under high levels of competition, which may help explain intra‐specific variation in competitive response. However, the relationships between traits and competitive response are not well understood, especially in trees. In this study, we investigated among‐genotype associations between tree trait plasticity and competitive response. We manipulated competition intensity in experimental stands of trembling aspen (Populus tremuloides) to address the covariance between competition‐induced changes in functional trait expression and aspects of competitive ability at the genotype level. Genotypic variation in the direction and magnitude of functional trait responses, especially those of crown foliar mass, phytochemistry, and leaf physiology, was associated with genotypic variation in competitive response. Traits exhibited distinct plastic responses to competition, with varying degrees of genotypic variation and covariance with other trait responses. The combination of genotypic diversity and covariance among functional traits led to tree responses to competition that were coordinated among traits yet variable among genotypes. Such relationships between tree traits and competitive success have the potential to shape stand‐level trait distributions over space and time.

Leaf-cutter ants are dominant neotropical herbivores capable of deriving energy from a wide range of plant substrates. The success of leaf-cutter ants is largely due to their external gut, composed of key microbial symbionts, specifically, the fungal mutualistL. gongylophorusand a consistent bacterial community. Both symbionts are known to have critical roles in extracting energy from plant material, yet comparatively little is known about their roles in the detoxification of plant secondary compounds. In this study, we assessed if the bacterial communities associated with leaf-cutter ant fungus gardens can degrade harmful plant chemicals. We identify plant secondary compound detoxification in leaf-cutter ant gardens as a process that depends on the degradative potential of both the bacterial community andL. gongylophorus. Our findings suggest that the fungus garden and its associated microbial community influence the generalist foraging abilities of the ants, underscoring the importance of microbial symbionts in plant substrate suitability for herbivores.