Biomechanical properties of non-flight vibrations produced by bees.

Bees use thoracic vibrations produced by their indirect flight muscles for powering wingbeats in flight, but also during mating, pollination, defence and nest building. Previous work on non-flight vibrations has mostly focused on acoustic (airborne vibrations) and spectral properties (frequency domain). However, mechanical properties such as the vibration's acceleration amplitude are important in some behaviours, e.g. during buzz pollination, where higher amplitude vibrations remove more pollen from flowers. Bee vibrations have been studied in only a handful of species and we know very little about how they vary among species. In this study, we conducted the largest survey to date of the biomechanical properties of non-flight bee buzzes. We focused on defence buzzes as they can be induced experimentally and provide a common currency to compare among taxa. We analysed 15,000 buzzes produced by 306 individuals in 65 species and six families from Mexico, Scotland and Australia. We found a strong association between body size and the acceleration amplitude of bee buzzes. Comparison of genera that buzz-pollinate and those that do not suggests that buzz-pollinating bees produce vibrations with higher acceleration amplitude. We found no relationship between bee size and the fundamental frequency of defence buzzes. Although our results suggest that body size is a major determinant of the amplitude of non-flight vibrations, we also observed considerable variation in vibration properties among bees of equivalent size and even within individuals. Both morphology and behaviour thus affect the biomechanical properties of non-flight buzzes.

Climatic and soil characteristics account for the genetic structure of the invasive cactus moth Cactoblastis cactorum, in its native range in Argentina.

Background: Knowledge of the physical and environmental conditions that may limit the migration of invasive species is crucial to assess the potential for expansion outside their native ranges. The cactus moth, Cactoblastis cactorum, is native to South America (Argentina, Paraguay, Uruguay and Brazil) and has been introduced and invaded the Caribbean and southern United States, among other regions. In North America there is an ongoing process of range expansion threatening cacti biodiversity of the genus Opuntia and the commercial profits of domesticated Opuntia ficus-indica. Methods: To further understand what influences the distribution and genetic structure of this otherwise important threat to native and managed ecosystems, in the present study we combined ecological niche modeling and population genetic analyses to identify potential environmental barriers in the native region of Argentina. Samples were collected on the host with the wider distribution range, O. ficus-indica. Results: Significant genetic structure was detected using 10 nuclear microsatellites and 24 sampling sites. At least six genetic groups delimited by mountain ranges, salt flats and wetlands were mainly located to the west of the Dry Chaco ecoregion. Niche modeling supports that this region has high environmental suitability where the upper soil temperature and humidity, soil carbon content and precipitation were the main environmental factors that explain the presence of the moth. Environmental filters such as the upper soil layer may be critical for pupal survival and consequently for the establishment of populations in new habitats, whereas the presence of available hosts is a necessary conditions for insect survival, upper soil and climatic characteristics will determine the opportunities for a successful establishment.

Darwin's inflorescence syndrome is indeed associated with bee pollination.

KEY MESSAGE: A relationship between vertical acropetal inflorescences with protandrous flowers and bee pollination was hypothesized by Darwin back in 1877. Here we provide empirical evidence supporting this association across the angiosperms. Plant reproduction is not only determined by flower traits but also by the arrangement of flowers within inflorescences. Based on his observations of the orchid Spiranthes autumnalis, Darwin proposed in 1877 that bee-pollinated plants presenting protandrous flowers on vertical acropetal inflorescences, where proximal flowers open first, can exploit the stereotypical foraging behavior of their pollinators (i.e., upward movement through the inflorescence) to promote pollen exportation and reduce self-pollination. In these inflorescences, male-phase flowers lie spatially above female-phase flowers. To examine this untested hypothesis, we compiled literature information from 718 angiosperms species and evaluated the association between vertical acropetal inflorescences with protandrous flowers and bee pollination within a phylogenetic comparative framework. Results reveal that this type of inflorescence is indeed more common in species pollinated by bees. Moreover, this association does not seem to be weakened by the presence of alternative self-pollination avoidance mechanisms, like self-incompatibility, suggesting that this inflorescence type benefits mainly male rather than female fitness. Other inflorescence types placing male-phase flowers above female-phase flowers, e.g., vertical basipetal inflorescences with protogynous flowers, do not provide strong evidence of a differential association with pollination by bees. Female-biased nectar production in vertical acropetal inflorescences with protandrous flowers may reinforce the behavior of bees to fly upwards, rendering Darwin's configuration more adaptive than other inflorescence configurations.

The role of within-plant variation in nectar production: an experimental approach.

BACKGROUND AND AIMS: Nectar, a plant reward for pollinators, can be energetically expensive. Hence, a higher investment in nectar production can lead to reduced allocation to other vital functions and/or increased geitonogamous pollination. One possible strategy employed by plants to reduce these costs is to offer variable amounts of nectar among flowers within a plant, to manipulate pollinator behaviour. Using artificial flowers, we tested this hypothesis by examining how pollinator visitation responds to inter- and intra-plant variation in nectar production, assessing how these responses impact the energetic cost per visit. METHODS: We conducted a 2 × 2 factorial experiment using artificial flowers, with two levels of nectar investment (high and low sugar concentration) and two degrees of intra-plant variation in nectar concentration (coefficient of variation 0 and 20 %). The experimental plants were exposed to visits (number and type) from a captive Bombus impatiens colony, and we recorded the total visitation rate, distinguishing geitonogamous from exogamous visits. Additionally, we calculated two estimators of the energetic cost per visit and examined whether flowers with higher nectar concentrations (richer flowers) attracted more bumblebees. KEY RESULTS: Plants in the variable nectar production treatment (coefficient of variation 20 %) had a greater proportion of flowers visited by pollinators, with higher rates of total, geitonogamous and exogamous visitation, compared with plants with invariable nectar production. When assuming no nectar reabsorption, variable plants incurred a lower cost per visit compared with invariable plants. Moreover, highly rewarding flowers on variable plants had higher rates of pollination visits compared with flowers with few rewards. CONCLUSIONS: Intra-plant variation in nectar concentration can represent a mechanism for pollinator manipulation, enabling plants to decrease the energetic costs of the interaction while still ensuring consistent pollinator visitation. However, our findings did not provide support for the hypothesis that intra-plant variation in nectar concentration acts as a mechanism to avoid geitonogamy. Additionally, our results confirmed the hypothesis that increased visitation to variable plants is dependent on the presence of flowers with nectar concentration above the mean.

Residual Effects of Transgenic Cotton on the Intestinal Microbiota of Dysdercus concinnus.

The interaction among plants, insects, and microbes (PIM) is a determinant factor for the assembly and functioning of natural and anthropic ecosystems. In agroecosystems, the relationships among PIM are based on the interacting taxa, environmental conditions, and agricultural management, including genetically modified (GM) organisms. Although evidence for the unintended effects of GM plants on non-target insects is increasingly robust, our knowledge remains limited regarding their impact on gut microbes and their repercussions on the host's ecology, especially in the wild. In this study, we compared the gut microbial community of Dysdercus concinnus bugs collected on wild cotton (Gossypium hirsutum), with and without insecticidal transgenes (cry1ab/ac), in its center of origin and diversity. By sequencing the V4-V5 region of 16S rRNA, we show differences in the diversity, structure, and topology of D. concinnus gut microbial interactions between specimens foraging cotton plants with and without transgenes. Identifying unintended residual effects of genetic engineering in natural ecosystems will provide first-line knowledge for informed decision-making to manage genetic, ecological, and evolutionary resources. Thus, determining which organisms interact with GM plants and how is the first step toward conserving natural ecosystems with evidence of transgenic introgression.

Did early shifts to bird pollination impose constraints on Salvia flower evolution?

A recent article by Kriebel et al. (2020) examines the relationship between floral shape evolution and pollination shifts in Salvia, a plant genus almost worldwide distributed, but particularly diverse in the New World. Kriebel et al. (2020) argue that a major shift to bird pollination at the origin of the subg. Calosphace (~20 million years ago [Mya]) imposed a legacy of constraints, resulting in significant differences in flower morphology between New World and Old World Salvia. However, reanalyses of the data using hidden states to account for the heterogeneity in evolutionary rates do not support an early origin of bird pollination in this group. Instead, bird pollination may have appeared after the arrival of modern hummingbirds to North America (15.5-12 Mya), as in other North American plant clades. The use of more complex models of ancestral state reconstruction into comparative analyses provides a different perspective to explain morphological differences within Salvia. Our results indicate that bird pollination did not impose constraints on corolla shape evolution. Evolutionary constraints in anther connective and style shapes may have arisen at the origin of Calosphace but they were not associated with shifts to hummingbird pollination, being more likely the product of contingent evolution.

Priming by Insects: Differential Effects of Sympatric and Allopatric Priming upon Plant Performance and Tolerance to Herbivory.

Plants have evolved multiple mechanisms to defend themselves from their multiple herbivores. Thus, being able to recognise among them and respond accordingly is fundamental for plant survival and reproduction. Defence priming prepares the plant to better or more rapidly respond to future damage; however, while it is considered an adaptive trait, to date, no studies have evaluated the extent and specificity of the priming recognition. To estimate the costs, benefits and specificity of priming, we used a highly specialist plant-insect system (Datura stramonium-Lema daturaphila) and performed a reciprocal transplant experiment with two populations where a priming stimulus (sympatric vs. allopatric) and a damage treatment (sympatric) were applied. We found no evidence of a fitness cost of priming, given that primed plants without damage showed no reduction in fitness. In contrast, our treatments affected the probability of bud abortion. That is, when damaged plants received no priming or the priming came from an allopatric insect, the likelihood of aborting the first bud was 1.9 times greater compared to plants being primed by their sympatric insect. We also found that damaged plants primed with an allopatric insect produced 14% fewer seeds compared to plants receiving a sympatric priming stimulus. Tolerance to herbivore damage was also the lowest when plants received the priming stimulus from an allopatric insect. Overall, these results suggest that, in our study system, plants recognise their local insect population reducing the negative effect of damage through a tolerance response.

Studying Plant-Insect Interactions through the Analyses of the Diversity, Composition, and Functional Inference of Their Bacteriomes.

As with many other trophic interactions, the interchange of microorganisms between plants and their herbivorous insects is unavoidable. To test the hypothesis that the composition and diversity of the insect bacteriome are driven by the bacteriome of the plant, the bacteriomes of both the plant Datura inoxia and its specialist insect Lema daturaphila were characterised using 16S sRNA gene amplicon sequencing. Specifically, the bacteriomes associated with seeds, leaves, eggs, guts, and frass were described and compared. Then, the functions of the most abundant bacterial lineages found in the samples were inferred. Finally, the patterns of co-abundance among both bacteriomes were determined following a multilayer network approach. In accordance with our hypothesis, most genera were shared between plants and insects, but their abundances differed significantly within the samples collected. In the insect tissues, the most abundant genera were Pseudomonas (24.64%) in the eggs, Serratia (88.46%) in the gut, and Pseudomonas (36.27%) in the frass. In contrast, the most abundant ones in the plant were Serratia (40%) in seeds, Serratia (67%) in foliar endophytes, and Hymenobacter (12.85%) in foliar epiphytes. Indeed, PERMANOVA analysis showed that the composition of the bacteriomes was clustered by sample type (F = 9.36, p < 0.001). Functional inferences relevant to the interaction showed that in the plant samples, the category of Biosynthesis of secondary metabolites was significantly abundant (1.4%). In turn, the category of Xenobiotics degradation and metabolism was significantly present (2.5%) in the insect samples. Finally, the phyla Proteobacteria and Actinobacteriota showed a pattern of co-abundance in the insect but not in the plant, suggesting that the co-abundance and not the presence−absence patterns might be more important when studying ecological interactions.

Local dispersal pathways during the invasion of the cactus moth, Cactoblastis cactorum, within North America and the Caribbean.

Cactoblastis cactorum, a species of moth native to Argentina, feeds on several prickly pear cactus species (Opuntia) and has been successfully used as a biological control of invading Opuntia species in Australia, South Africa and native ruderal Opuntia species in some Caribbean islands. Since its introduction to the Caribbean its spread was uncontrolled, invading successfully Florida, Texas and Louisiana. Despite this long history of invasion, we are still far from understanding the factors determining the patterns of invasion of Cactoblastis in North America. Here, we explored three non-mutually exclusive explanations: a) a stepping stone model of colonization, b) long distance colonization due to hurricanes, and/or c) hitchhiking through previously reported commercial routes. Genetic diversity, genetic structure and the patterns of migration among populations were obtained by analyzing 10 nuclear microsatellite loci. Results revealed the presence of genetic structure among populations of C. cactorum in the invaded region and suggest that both marine commercial trade between the Caribbean islands and continental USA, as well as recurrent transport by hurricanes, explain the observed patterns of colonization. Provided that sanitary regulations avoiding human-mediated dispersal are enforced, hurricanes probably represent the most important agent of dispersal and future invasion to continental areas.

Ontogenetic changes in the targets of natural selection in three plant defenses.

The evolution of plant defenses has traditionally been studied at single plant ontogenetic stages, overlooking the fact that natural selection acts continuously on organisms along their development, and that the adaptive value of phenotypes can change along ontogeny. We exposed 20 replicated genotypes of Turnera velutina to field conditions to evaluate whether the targets of natural selection on different defenses and their adaptative value change across plant development. We found that low chemical defense was favored in seedlings, which seems to be explained by the assimilation efficiency and the ability of the specialist herbivore to sequester cyanogenic glycosides. Whereas trichome density was unfavored in juvenile plants, it increased relative plant fitness in reproductive plants. At this stage we also found a positive correlative gradient between cyanogenic potential and sugar content in extrafloral nectar. We visualize this complex multi-trait combination as an ontogenetic defensive strategy. The inclusion of whole-plant ontogeny as a key source of variation in plant defense revealed that the targets and intensity of selection change along the development of plants, indicating that the influence of natural selection cannot be inferred without the assessment of ontogenetic strategies in the expression of multiple defenses.

Natural selection acting on integrated phenotypes: covariance among functional leaf traits increases plant fitness.

Plant functional strategies are usually accomplished through the simultaneous expression of different traits, and hence their correlations should be promoted by natural selection. The adaptive value of correlations among leaf functional traits, however, has not been assessed in natural populations. We estimated intraspecific variation in leaf functional traits related to the primary metabolism and anti-herbivore defence in a population of Turnera velutina. We analysed whether natural selection favoured the expression of individual traits, particular combinations of traits or leaf phenotypic integration. Patterns of covariation among traits were related to water and nitrogen economy, and were similar among genotypes, but the magnitude of their phenotypic integration differed by 10-fold. Although families did not differ in the mean values of leaf functional traits, directional selection favoured low nitrogen content and low chemical defence, high content of chlorophyll, sugar in extrafloral nectar and trichome density. Families with higher phenotypic integration among leaf traits grew faster and produced more flowers. We suggest that the coordinated expression of leaf traits has an adaptive value, probably related to optimisation in the expression of traits related to water conservation and nitrogen acquisition.

Daily fluctuations in pollination effectiveness explain higher efficiency of native over exotic bees in Lepechinia floribunda (Lamiaceae).

BACKGROUND: Despite Stebbins' principle of the most efficient pollinator being proposed decades ago, the most important pollinators are still mainly identified using the frequency of visits to flowers. This shortcoming results in a gap between the characterization of the flower visitors of a plant species and a reliable estimation of the plant fitness consequences of the mutualistic interaction. The performance of a mutualistic visitor depends on its abundance, behaviour, effectiveness (pollen removal and deposition per unit time) and efficiency (seed set per unit time) conditioned by the temporal matching between pollinator activity and temporal patterns of maturation of the sexual functions of flowers. Although there have been recent attempts to provide a conceptual and methodological framework to characterize pollinators' performance, few have combined all key elements of visitors and plants to provide an accurate estimation of pollinators' performance under natural conditions. METHODS: We complement information on the flower biology and mating system of the sub-shrub Lepechinia floribunda (Lamiaceae) to provide a daily quantitative estimation of performance (effectiveness and efficiency) of the more abundant pollinators, i.e. native bumble-bees (Bombus spp.) and leafcutter bees (Megachile sp.), and the exotic honey-bee (Apis mellifera). KEY RESULTS: Unlike honey-bees or leafcutter bees, native bumble-bees matched the daily pattern of nectar production and stigma receptivity, and showed higher effectiveness and efficiency. Despite the overabundance of honey-bees, visits occurred mainly when stigmas were not receptive, thus reducing the honey-bees' overall performance. CONCLUSIONS: Bumble-bees appear to be the most important pollinators and potential historical mediators of reproductive trait evolution in L. floribunda. Because the production of seeds by bumble-bees involved fewer pollen grains for plants and less investment in floral display than honey-bees, contemporary and expected changes in pollinator abundance may affect future L. floribunda floral evolution. If bumble-bees were to be further displaced by anthropogenic disturbance or by competition with honey-bees, their lower efficiency will select for a larger floral display increasing reproductive costs. This scenario may also impose selection to reduce dichogamy to match honey-bee foraging activity.

Ontogenetic trajectories of direct and indirect defenses of myrmecophytic plants colonized either by mutualistic or opportunistic ant species.

Myrmecophytic plants are expected to produce greater direct defenses when young and switch towards indirect defenses once they reach the size and vigor to produce enough rewards for their ant mutualists. The presence of opportunistic ant species, however, is likely to promote the variation in these ontogenetic trajectories. When plants do not obtain benefits from ants, they cannot rely on this indirect defense. Hence, the expression of direct defenses is expected to remain constant or even increase during the development of plants colonized by opportunistic ants, whereas a reduction in resource allocation to indirect defenses should be observed. To assess if myrmecophytic plants adjust their ontogenetic trajectories in defense as a function of the colonizing ant species, we estimated direct and indirect defenses at four ontogenetic stages of the myrmecophytic plant Vachellia hindsii colonized by either mutualistic or opportunistic ant partners. We report that cyanogenic potential decreased while leaf thickness and the production of sugar in extrafloral nectaries increased along plant development. The magnitude of these ontogenetic changes, however, varied as a function of the identity of the colonizing ants. As expected, when colonized by opportunistic ants, plants produced more direct defenses and reduced the production of rewards. We suggest that facultative changes in the expression of ontogenetic trajectories in direct and indirect defenses could be a mechanism to reduce the fitness costs associated with opportunistic interactions.

Evolutionary transition between bee pollination and hummingbird pollination in Salvia: Comparing means, variances and covariances of corolla traits.

Covariation among traits can modify the evolutionary trajectory of complex structures. This process is thought to operate at a microevolutionary scale, but its long-term effects remain controversial because trait covariation can itself evolve. Flower morphology, and particularly floral trait (co)variation, has been envisioned as the product of pollinator-mediated selection. Available evidence suggests that major changes in pollinator assemblages may affect the joint expression of floral traits and their phenotypic integration. We expect species within a monophyletic lineage sharing the same pollinator type will show not only similarity in trait means but also similar phenotypic variance-covariance structures. Here, we tested this expectation using eighteen Salvia species pollinated either by bees or by hummingbirds. Our findings indicated a nonsignificant multivariate phylogenetic signal and a decoupling between means and variance-covariance phenotypic matrices of floral traits during the evolution to hummingbird pollination. Mean trait value analyses revealed significant differences between bee- and hummingbird-pollinated Salvia species although fewer differences were detected in the covariance structure between groups. Variance-covariance matrices were much more similar among bee- than hummingbird-pollinated species. This pattern is consistent with the expectation that, unlike hummingbirds, bees physically manipulate the flower, presumably exerting stronger selection pressures favouring morphological convergence among species. Overall, we conclude that the evolution of hummingbird pollination proceeded through different independent transitions. Thus, although the evolution of hummingbird pollination led to a new phenotypic optimum, the process involved the diversification of the covariance structure.

On the adaptive value of monomorphic versus dimorphic enantiostyly in Solanum rostratum.

Background and Aims: Enantiostyly is a reproductive system with heteromorphic flowers characterized by asymmetrical deflection of the style, either to the left or to the right of the floral axis. There are two types of enantiostyly. In monomorphic enantiostyly, plants produce the two types of flowers in the same individual. Dimorphic enantiostyly is restricted to only seven species and their populations consist of individuals producing either the right or the left flower type. It is hypothesized that the dimorphic form is derived from monomorphic ancestors because it functions as an outcrossing mechanism. We tested this latter hypothesis and investigated if monomorphic enantiostyly is resistant to invasion by individuals with dimorphic enantiostyly, because it functions as a reproductive assurance mechanism. Methods: To determine the conditions favouring the invasion of dimorphic enantiostyly, measurements of reproductive success and outcrossing rates in 15 natural flowering patches of Solanum rostratum were made. To test if monomorphic enantiostyly provides a reproductive assurance mechanism, experimental plants with either manually created dimorphic or natural monomorphic reproductive systems were exposed to two different pollination scenarios (flower density treatments), and reproductive success and outcrossing rates were measured. Key Results: Naturally flowering patches experienced severe pollination limitation, showed marked differences in reproductive success and had relatively high outcrossing rates. Plants in the experimental patches also showed pollination limitation and high outcrossing rates. Individuals with dimorphic enantiostyly expressed higher reproductive and outcrossing advantages under high-density conditions. These advantages disappeared in the low-density treatment, where the monomorphic form attained a higher reproductive success and no differences in outcrossing rates were detected. Conclusions: Monomorphic enantiostyly should be resistant to invasion of the dimorphic form because the prevalent ecological conditions favour the maintenance of geitonogamous individuals that are able to take advantage of ecological heterogeneity and generalized pollination limitation.

Risk of herbivore attack and heritability of ontogenetic trajectories in plant defense.

Ontogeny has been identified as a main source of variation in the expression of plant phenotypes. However, there is limited information on the mechanisms behind the evolution of ontogenetic trajectories in plant defense. We explored if risk of attack, herbivore damage, heritability, and phenotypic plasticity can promote or constrain the evolutionary potential of ontogenetic trajectories in three defensive traits. We exposed 20 genotypes of Turnera velutina to contrasting environments (shadehouse and field plots), and measured the cyanogenic potential, trichome density, and sugar content in extrafloral nectar in seedlings, juveniles and reproductive plants. We also assessed risk of attack through oviposition preferences, and quantified herbivore damage in the field. We estimated genetic variance, broad sense heritability, and evolvability of the defensive traits at each ontogenetic stage, and of the ontogenetic trajectories themselves. For plants growing in the shadehouse, we found genetic variation and broad sense heritability for cyanogenic potential in seedlings, and for trichome density at all ontogenetic stages. Genetic variation and heritability of ontogenetic trajectories was detected for trichome density only. These genetic pre-requisites for evolution, however, were not detected in the field, suggesting that environmental variation and phenotypic plastic responses mask any heritable variation. Finally, ontogenetic trajectories were found to be plastic, differing between shadehouse and field conditions for the same genetic families. Overall, we provide support for the idea that changes in herbivore pressure can be a mechanism behind the evolution of ontogenetic trajectories. This evolutionary potential, however, can be constrained by phenotypic plasticity expressed in heterogeneous environments.

The evolution of floral ontogenetic allometry in the Andean genus Caiophora (Loasaceae, subfam. Loasoideae).

The astounding variety of angiosperm flower morphologies has evolved in response to many selective forces. Flower development is highly coordinated and involves developmental associations between size and shape, ontogenetic allometry, which in turn affect the morphology of mature flowers. Although ontogenetic allometries can act as a developmental constraint and may influence adaptive evolution, allometries can evolve themselves and may change rapidly in response to selection. We explored the evolution of ontogenetic allometry in the flowers of 11 species of Loasoideae. Seven species belong to Caiophora, which radiated recently in the central Andes, and contains species that are pollinated by bees, hummingbirds, and small rodents. According to a previous study, the diversification of Caiophora involved departures from simple allometric scaling, but the changes to allometry that enabled flower diversification have not been explored yet. We characterized the ontogenetic allometry of each species with the methods of geometric morphometrics. We studied the evolution of allometries by constructing allometric spaces, in which the allometry of each species is represented by a point and the arrangement of points indicates the relations among allometric trajectories. To examine the history of changes of ontogenetic allometries, we projected the phylogeny into the allometric spaces. Inspection of allometric spaces suggests that ontogenetic variation is limited to a few dominant features. The allometries of the two main functional flower parts under study differ in their evolutionary labilities, and patterns of variation reflect pollination systems, differences in structural organization, and abiotic environmental factors.

Testing the hypothesis that biological modularity is shaped by adaptation: Xylem in the Bursera simaruba clade of tropical trees.

The study of modularity allows recognition of suites of character covariation that potentially diagnose units of evolutionary change. One prominent perspective predicts that natural selection should forge developmental units that maximize mutual functional independence. We examined the module-function relation using secondary xylem (wood) in a clade of tropical trees as a study system. Traditionally, the three main cell types in wood (vessels, fibers, and parenchyma) have respectively been associated with three functions (conduction, mechanical support, and storage). We collected samples from nine species of the simaruba clade of Bursera at fifteen sites and measured thirteen anatomical variables that have traditionally been regarded as reflecting the distinct functions of these cell types. If there are indeed (semi) independently evolving modules associated with functions, and cell types really are associated with these functions, then we should observe greater association between traits within cell types than between traits from different cell types. To map these associations, we calculated correlation coefficients among anatomical variables and identified modules using cluster and factor analysis. Our results were only partially congruent with expectations, with associations between characters of different cell types common. These results suggest causes of covariation, some involving selected function as predicted, but also highlighting the tradeoffs and shared developmental pathways limiting the evolutionary independence of some cell types in the secondary xylem. The evolution of diversity across the simaruba clade appears to have required only limited independence between parts.

Exploring the ontogenetic scaling hypothesis during the diversification of pollination syndromes in Caiophora (Loasaceae, subfam. Loasoideae).

BACKGROUND AND AIMS: Phenotypic diversification of flowers is frequently attributed to selection by different functional groups of pollinators. During optimization of floral phenotype, developmental robustness to genetic and non-genetic perturbations is expected to limit the phenotypic space available for future evolutionary changes. Although adaptive divergence can occur without altering the basic developmental programme of the flower (ontogenetic scaling hypothesis), the rarity of reversion to ancestral states following adaptive radiations of pollination syndromes suggests that changes in the ancestral developmental programme of the flower are common during such evolutionary transitions. Evidence suggests that flower diversification into different pollination syndromes in the Loasoideae genus Caiophora took place during a recent adaptive radiation in the central Andes. This involved transitions from bee to hummingbird and small rodent pollination. The aim of this work was to examine if the adaptive radiation of pollination syndromes in Caiophora occurred through ontogenetic scaling or involved a departure from the ontogenetic pattern basal to this genus. METHODS: We used geometric morphometric variables to describe the shape and size of floral structures taking part in the pollination mechanism of Loasoideae. This approach was used to characterize the developmental trajectories of three species basal to the genus Caiophora through shape-size relationships (ontogenetic allometry). We then tested if the shape-size combinations of these structures in mature flowers of derived Caiophora species fall within the phenotypic space predicted by the development of basal species. KEY RESULTS: Variation in the size and shape of Caiophora flowers does not overlap with the pattern of ontogenetic allometry of basal species. Derived bee-, hummingbird- and rodent-pollinated species had divergent ontogenetic patterns of floral development from that observed for basal bee-pollinated species. CONCLUSIONS: The adaptive radiation of Caiophora involved significant changes in the developmental pattern of the flowers, rejecting the ontogenetic scaling hypothesis.

Natural selection drives chemical resistance of Datura stramonium.

Plant resistance to herbivores involves physical and chemical plant traits that prevent or diminish damage by herbivores, and hence may promote coevolutionary arm-races between interacting species. Although Datura stramonium's concentration of tropane alkaloids is under selection by leaf beetles, it is not known whether chemical defense reduces seed predation by the specialist weevil, Trichobaris soror, and if it is evolving by natural selection. We measured infestation by T. soror as well as the concentration of the plants' two main tropane alkaloids in 278 D. stramonium plants belonging to 31 populations in central Mexico. We assessed whether the seed predator exerted preferences on the levels of both alkaloids and whether they affect plant fitness. Results show great variation across populations in the concentration of scopolamine and atropine in both leaves and seeds of plants of D. stramonium, as well as in the intensity of infestation and the proportion of infested fruits by T. soror. The concentration of scopolamine in seeds and leaves are negatively associated across populations. We found that scopolamine concentration increases plant fitness. Our major finding was the detection of a positive relationship between the population average concentrations of scopolamine with the selection differentials of scopolamine. Such spatial variation in the direction and intensity of selection on scopolamine may represent a coevolutionary selective mosaic. Our results support the view that variation in the concentration of scopolamine among-populations of D. stramonium in central Mexico is being driven, in part, by selection exerted by T. soror, pointing an adaptive role of tropane alkaloids in this plant species.