Multifunctionality of angiosperm floral bracts: a review.

Floral bracts (bracteoles, cataphylls) are leaf-like organs that subtend flowers or inflorescences but are of non-floral origin; they occur in a wide diversity of species, representing multiple independent origins, and exhibit great variation in form and function. Although much attention has been paid to bracts over the past 150 years, our understanding of their adaptive significance remains remarkably incomplete. This is because most studies of bract function and evolution focus on only one or a few selective factors. It is widely recognised that bracts experience selection mediated by pollinators, particularly for enhancing pollinator attraction through strong visual, olfactory, or echo-acoustic contrast with the background and through signalling the presence of pollinator rewards, either honestly (providing rewards for pollinators), or deceptively (attraction without reward or even trapping pollinators). However, studies in recent decades have demonstrated that bract evolution is also affected by agents other than pollinators. Bracts can protect flowers, fruits, or seeds from herbivores by displaying warning signals, camouflaging conspicuous reproductive organs, or by providing physical barriers or toxic chemicals. Reviews of published studies show that bracts can also promote seed dispersal and ameliorate the effects of abiotic stressors, such as low temperature, strong ultraviolet radiation, heavy rain, drought, and/or mechanical abrasion, on reproductive organs or for the plants' pollinators. In addition, green bracts and greening of colourful bracts after pollination promote photosynthetic activity, providing substantial carbon (photosynthates) for fruit or seed development, especially late in a plant's life cycle or season, when leaves have started to senesce. A further layer of complexity derives from the fact that the agents of selection driving the evolution of bracts vary between species and even between different developmental stages within a species, and selection by one agent can be reinforced or opposed by other agents. In summary, our survey of the literature reveals that bracts are multifunctional and subject to multiple agents of selection. To understand fully the functional and evolutionary significance of bracts, it is necessary to consider multiple selection agents throughout the life of the plant, using integrative approaches to data collection and analysis.

Incomplete partitioning of pollinators by Linum suffruticosum and its coflowering congeners.

PREMISE: Linum suffruticosum shows variations in pollinator fit, pollen pickup, and local pollinators that predict pollen deposition rates. The species often coflowers with other Linum species using the same pollinators. We investigated whether L. suffruticosum trait variation could be explained by local patterns of pollinator sharing and associated evolution to reduce interspecific pollen transfer. METHODS: Pollinator observations were made in different localities (single species, coflowering with other congeners). Floral traits were measured to detect differences across populations and from coflowering species. Reproductive costs were quantified using interspecific hand pollinations and measures of pollen-tube formation, combined with observations of pollen arrival on stigmas and pollen-tube formation after natural pollination in allopatric and sympatric localities. RESULTS: The size and identity of the most important pollinator of L. suffruticosum and whether there was pollinator sharing with coflowering species appeared to explain floral trait variation related to pollinator fit. The morphological overlap of the flowers of L. suffruticosum with those of coflowering species varied, depending on coflowering species identity. A post-pollination incompatibility system maintains reproductive isolation, but conspecific pollen-tube formation was lower after heterospecific pollination. Under natural pollination at sites of coflowering with congeners, conspecific pollen-tube formation was lower than at single-species localities. CONCLUSIONS: Trait variation in L. suffruticosum appears to respond to the most important local pollinator. Locally, incomplete pollinator partitioning might cause interspecific pollination, imposing reproductive costs. These reproductive costs may generate selection on floral traits for reduced morphological overlap with coflowering congeners, leading to the evolution of pollination ecotypes.

Plant ecology: New insights into the adaptive significance of rapid floral movements.

Rapid movements by plants have been a subject of interest since Linnaeus in the 18th century. Largely lacking, however, has been a modern understanding of the adaptive significance of such movements. A recent study shows that rapid stamen movements can increase male fitness by scaring off pollinators once they have pollen placed on them.

Evolvability and trait function predict phenotypic divergence of plant populations.

Understanding the causes and limits of population divergence in phenotypic traits is a fundamental aim of evolutionary biology, with the potential to yield predictions of adaptation to environmental change. Reciprocal transplant experiments and the evaluation of optimality models suggest that local adaptation is common but not universal, and some studies suggest that trait divergence is highly constrained by genetic variances and covariances of complex phenotypes. We analyze a large database of population divergence in plants and evaluate whether evolutionary divergence scales positively with standing genetic variation within populations (evolvability), as expected if genetic constraints are evolutionarily important. We further evaluate differences in divergence and evolvability-divergence relationships between reproductive and vegetative traits and between selfing, mixed-mating, and outcrossing species, as these factors are expected to influence both patterns of selection and evolutionary potentials. Evolutionary divergence scaled positively with evolvability. Furthermore, trait divergence was greater for vegetative traits than for floral (reproductive) traits, but largely independent of the mating system. Jointly, these factors explained ~40% of the variance in evolutionary divergence. The consistency of the evolvability-divergence relationships across diverse species suggests substantial predictability of trait divergence. The results are also consistent with genetic constraints playing a role in evolutionary divergence.

Degree of style coiling is associated with corolla-tube length in the nectarless flowers of Roscoea schneideriana.

The plant-pollinator 'arms race' model posits that a major driver of the evolution of elongated corollas in flowers is reciprocal selection for 'morphological fit' between pollinator-tongue length and access distance to nectar (usually corolla-tube length). Evidence for the pollinator-mediated selection on tube length and evolution of multiple, correlated floral traits remains inconclusive. To gain possible insights into the strength of stabilizing selection by assessing standing phenotypic variation, we measured a series of functionally important floral traits, including corolla tube length and 'effective' tube depth and degree of style coiling. We then calculated coefficients of variation (CV) for these traits in three field populations of R. schneideriana. Unlike in most long-tubed flowers, the bottom part of the corolla tube is completely occupied by the style, with no room for nectar. The length of this portion of the corolla tube was more variable (higher CV) than the upper part of the corolla tube, suggesting that functional tube depth was under stronger stabilizing selection. The degree of style coiling was negatively related to the corolla-tube length in all three populations of R. schneideriana, suggesting that there may be conflicting selection acting on style length and corolla-tube length, which are otherwise usually tightly correlated. Given the lack of nectar in the flowers of this species, the long corolla tubes and long styles may represent morphological holdovers from ancestors that were pollinated by long-tongued pollinators, as is still seen in related species in the western Himalayas.

Evolvability and constraint in the evolution of three-dimensional flower morphology.

PREMISE: Flower phenotypes evolve to attract pollinators and to ensure efficient pollen transfer to and from the bodies of pollinators or, in self-compatible bisexual flowers, between anthers and stigmas. If functionally interacting traits are genetically correlated, response to selection may be subject to genetic constraints. Genetic constraints can be assessed by quantifying standing genetic variation in (multivariate) phenotypic traits and by asking how much the available variation is reduced under specific assumptions about phenotypic selection on functionally interacting and genetically correlated traits. METHODS: We evaluated multivariate evolvability and potential genetic constraints underlying the evolution of the three-dimensional structure of Dalechampia blossoms. First, we used data from a greenhouse crossing design to estimate the G matrix for traits representing the relative positions of male and female sexual organs (anthers and stigmas) and used the G matrix to ask how genetic variation is distributed in multivariate space. To assess the evolutionary importance of genetic constraints, we related standing genetic variation across phenotypic space to evolutionary divergence of population and species in the same phenotypic directions. RESULTS: Evolvabilities varied substantially across phenotype space, suggesting that certain traits or trait combinations may be subject to strong genetic constraint. Traits involved functionally in flower-pollinator fit and autonomous selfing exhibited considerable independent evolutionary potential, but population and species divergence tended to occur in phenotypic directions associated with greater-than-average evolvability. CONCLUSIONS: These results are consistent with the hypothesis that genetic constraints can hamper joint trait evolution towards optimum flower-pollinator fit and optimum self-pollination rates.

Global analysis of floral longevity reveals latitudinal gradients and biotic and abiotic correlates.

The length of time a flower remains open and functional - floral longevity - governs important reproductive processes influencing pollination and mating and varies considerably among angiosperm species. However, little is known about large-scale biogeographic patterns and the correlates of floral longevity. Using published data on floral longevity from 818 angiosperm species in 134 families and 472 locations world-wide, we present the first global quantification of the latitudinal pattern of floral longevity and the relationships between floral longevity and a range of biotic and abiotic factors. Floral longevity exhibited a significant phylogenetic signal and was longer at higher latitudes in both northern and southern hemispheres, even after accounting for elevation. This latitudinal variation was associated with several biotic and abiotic variables. The mean temperature of the flowering season had the highest predictive power for floral longevity, followed by pollen number per flower. Surprisingly, compatibility status, flower size, pollination mode, and growth form had no significant effects on flower longevity. Our results suggest that physiological processes associated with floral maintenance play a key role in explaining latitudinal variation in floral longevity across global ecosystems, with potential implications for floral longevity under global climate change and species distributions.

Quantitative assessment of observed versus predicted responses to selection.

Although artificial-selection experiments seem well suited to testing our ability to predict evolution, the correspondence between predicted and observed responses is often ambiguous due to the lack of uncertainty estimates. We present equations for assessing prediction error in direct and indirect responses to selection that integrate uncertainty in genetic parameters used for prediction and sampling effects during selection. Using these, we analyzed a selection experiment on floral traits replicated in two taxa of the Dalechampia scandens (Euphorbiaceae) species complex for which G-matrices were obtained from a diallel breeding design. After four episodes of bidirectional selection, direct and indirect responses remained within wide prediction intervals, but appeared different from the predictions. Combined analyses with structural-equation models confirmed that responses were asymmetrical and lower than predicted in both species. We show that genetic drift is likely to be a dominant source of uncertainty in typically-dimensioned selection experiments in plants and a major obstacle to predicting short-term evolutionary trajectories.

Using ecological context to interpret spatiotemporal variation in natural selection.

Spatiotemporal variation in natural selection is expected, but difficult to estimate. Pollinator-mediated selection on floral traits provides a good system for understanding and linking variation in selection to differences in ecological context. We studied pollinator-mediated selection in five populations of Dalechampia scandens (Euphorbiaceae) in Costa Rica and Mexico. Using a nonlinear path-analytical approach, we assessed several functional components of selection, and linked variation in pollinator-mediated selection across time and space to variation in pollinator assemblages. After correcting for estimation error, we detected moderate variation in net selection on two out of four blossom traits. Both the opportunity for selection and the mean strength of selection decreased with increasing reliability of cross-pollination. Selection for pollinator attraction was consistently positive and stronger on advertisement than reward traits. Selection on traits affecting pollen transfer from the pollinator to the stigmas was strong only when cross-pollination was unreliable and there was a mismatch between pollinator and blossom size. These results illustrate how consideration of trait function and ecological context can facilitate both the detection and the causal understanding of spatiotemporal variation in natural selection.

Floral reorientation: the restoration of pollination accuracy after accidents.

Plants sometimes suffer mechanical injury. The nonlethal collapse of a flowering stalk, for example, can greatly reduce plant fitness if it leads to 'incorrect' floral orientation and thus reduced visitation or poor pollination. When floral orientation is important for accurate pollination, as has been suggested for bilaterally symmetrical flowers, we predict that such flowers should have developmental and/or behavioural mechanisms for restoring 'correct' orientation after accidents. We made observations and conducted experiments on 23 native and cultivated flowering plant species in Australia, South America, North America and Europe. We found that flowers with bilateral symmetry usually have the capacity to reorient after accidents, and that this is manifested through rapid bending and/or rotation of pedicels or sexual organs or slower peduncle bending. Floral reorientation restores pollination accuracy and fit with pollinators. However, experimental floral misorientation in eight species with radially symmetrical flowers showed that, with one exception, they had little capacity to reorient their flowers, in line with expectations that the orientation of radially symmetrical flowers does not substantially affect pollination accuracy. Our results suggest that quick corrective reorientation of bilaterally symmetrical flowers is adaptive, highlighting a little-studied aspect of plant-pollinator interactions and plant evolution.

Erratum: Author Correction: Modularity increases rate of floral evolution and adaptive success for functionally specialized pollination systems.

[This corrects the article DOI: 10.1038/s42003-019-0697-7.].

Fitness costs of delayed pollination in a mixed-mating plant.

BACKGROUND AND AIMS: To predict the evolutionary consequences of pollinator declines, we need to understand the evolution of delayed autonomous self-pollination, which is expected to evolve as a mechanism of reproductive assurance when cross-pollination becomes unreliable. This involves estimating the costs of increased levels of selfing as well as those associated with floral senescence. METHODS: We studied the mechanisms and costs of delayed self-pollination in the mixed-mating vine Dalechampia scandens (Euphorbiaceae) by first assessing among-population variation in herkogamy and dichogamy, which together determine the rate and timing of autonomous self-pollination. We then tested whether floral longevity responds plastically to delayed pollination. Finally, we assessed the costs of delayed self-pollination in terms of seed number and size, explicitly separating inbreeding depression from effects of floral senescence. KEY RESULTS: Herkogamy varied extensively, while variation in dichogamy was more limited. Unpollinated blossoms increased their longevity, but seed quantity and quality decreased with increasing delays in pollination, independently of inbreeding depression. CONCLUSIONS: In D. scandens, earlier autonomous selfing is facilitated by reduced herkogamy rather than reduced protogyny, providing reproductive assurance while maintaining the possibility for outcrossing events. Effective early autonomous self-pollination may evolve under reduced cross-pollination reliability in response to costs associated with floral senescence.

No evidence that seed predators constrain pollinator-mediated trait evolution in a tropical vine.

PREMISE OF THE STUDY: Turnover in biotic communities across heterogeneous landscapes is expected to lead to variation in interactions among plants, their mutualists, and their antagonists. Across a fragmented landscape in northern Costa Rica, populations of the euphorb vine Dalechampia scandens vary widely in mating systems and associated blossom traits. Previous work suggested that populations are well adapted to the local reliability of pollination by apid and megachilid bees. We tested whether variation in the intensity of predispersal seed predation by seed weevils in the genus Nanobaris also contributes to the observed variation in blossom traits. METHODS: We studied spatiotemporal variation in the relationships between floral advertisement and the probability of seed predation within three focal populations. Then we assessed among-population covariation of predation rate, pollination reliability, mating system, and blossom traits across 20 populations. KEY RESULTS: The probability of seed predation was largely unrelated to variation in floral advertisement both within focal populations and among the larger sample of populations. The rate of seed predation was only weakly associated with the rate of cross-pollination (allogamy) in each population but tended to be proportionally greater in populations experiencing less reliable pollination. CONCLUSIONS: These results suggest that geographic variation in the intensity of antagonistic interactions have had only minor modifying effects on the evolutionary trajectories of floral advertisement in plant populations in this system. Thus, pollinator-driven floral trait evolution in D. scandens in the study area appears not to be influenced by conflicting seed-predator-mediated selection.

Quantitative and qualitative consequences of reduced pollen loads in a mixed-mating plant.

Greater pollination intensity can enhance maternal plant fitness by increasing seed set and seed quality as a result of more intense pollen competition or enhanced genetic sampling. We tested experimentally these effects by varying the pollen load from a single pollen donor on stigmas of female flowers of Dalechampia scandens (Euphorbiaceae) and measuring the effects on seed number and seed mass. Seed set increased rapidly with pollen number at low to moderate pollen loads, and a maximum set of three seeds occurred with a mean pollen load of 19 pollen grains. We did not detect a trade-off between the number of seeds and seed mass within a fruit. Seed mass increased with increasing pollen load, supporting the hypothesis of enhanced seed quality via increased pollen-competition intensity or genetic sampling. These results suggest that maternal fitness increases with larger pollen loads, even when the fertilization success is already high. Our results further highlight the importance of high rates of pollen arrival onto stigmas, as mediated by reliable pollinators. Comparing the pollen-to-seed response curve obtained in this experiment with those observed in natural populations suggests that pollen limitation may be more severe in natural populations than predicted from greenhouse studies. These results also indicate that declines in pollinator abundance may decrease plant fitness through lowered seed quality before an effect on seed set is detected.

Detecting canalization and intra-floral modularity in triggerplant (Stylidium) flowers: correlations are only part of the story.

Background and Aims: The Berg hypothesis proposes that specialized-flower traits experience stronger stabilizing selection than non-floral structures and predicts that variation in specialized-flower traits will be mostly uncorrelated with variation in non-floral traits. Similarly, adaptive-accuracy theory predicts lower variation (as a proportion of the mean) in floral traits than in non-floral ones. Both hypotheses can be extended to comparisons between floral traits, where different parts of the flower can be expected to experience different strengths of stabilizing selection, resulting in contrasting patterns of variation. The present study tests these ideas by analysing variation/covariation in those floral traits influencing the location of pollen placement on, and stigma contact with, pollinators ('pollination-mechanics traits', PMTs) in relation to variation/covariation in non-floral traits and floral traits not directly involved in the mechanics of pollination. The prediction was that PMTs are canalized (buffered against genetic and environmental variation) relative to attraction traits, as manifested in lower variances and modular independence. Methods: Floral and inflorescence structures of ten species of triggerplants (Stylidium, Stylidiaceae) in south-western Australia were measured; the data were analysed using multivariate and bivariate approaches to detect modular structure of floral and non-floral traits and assess evidence for canalization of PMTs. Key Results: Only six of the ten species had PMTs with smaller correlation coefficients than attraction traits, in contrast to the Berg expectation. However, allometric and variance patterns were generally consistent with the predictions of an extended Berg hypothesis and adaptive accuracy. There was modular separation of most floral traits from non-floral traits and clear intra-floral modular structure. PMTs showed lower proportional variation and shallower allometric slopes than pollinator-attraction traits in nine and eight, respectively, of ten species. Conclusions: This study demonstrates the value of allometric and variance analyses (in addition to correlation) in assessing the evolutionary significance of floral-trait stability and plasticity.

Beyond buzz-pollination - departures from an adaptive plateau lead to new pollination syndromes.

Pollination syndromes describe recurring adaptation to selection imposed by distinct pollinators. We tested for pollination syndromes in Merianieae (Melastomataceae), which contain bee- (buzz-), hummingbird-, flowerpiercer-, passerine-, bat- and rodent-pollinated species. Further, we explored trait changes correlated with the repeated shifts away from buzz-pollination, which represents an 'adaptive plateau' in Melastomataceae. We used random forest analyses to identify key traits associated with the different pollinators of 19 Merianieae species and estimated the pollination syndromes of 42 more species. We employed morphospace analyses to compare the morphological diversity (disparity) among syndromes. We identified three pollination syndromes ('buzz-bee', 'mixed-vertebrate' and 'passerine'), characterized by different pollen expulsion mechanisms and reward types, but not by traditional syndrome characters. Further, we found that 'efficiency' rather than 'attraction' traits were important for syndrome circumscription. Contrary to syndrome theory, our study supports the pooling of different pollinators (hummingbirds, bats, rodents and flowerpiercers) into the 'mixed-vertebrate' syndrome, and we found that disparity was highest in the 'buzz-bee' syndrome. We conclude that the highly adaptive buzz-pollination system may have prevented shifts towards classical pollination syndromes, but provided the starting point for the evolution of a novel set of distinct syndromes, all having retained multifunctional stamens that provide pollen expulsion, reward and attraction.

Intersexual conflict over seed size is stronger in more outcrossed populations of a mixed-mating plant.

In polyandrous species, fathers benefit from attracting greater maternal investment toward their offspring at the expense of the offspring of other males, while mothers should usually allocate resources equally among offspring. This conflict can lead to an evolutionary arms race between the sexes, manifested through antagonistic genes whose expression in offspring depends upon the parent of origin. The arms race may involve an increase in the strength of maternally versus paternally derived alleles engaged in a "tug of war" over maternal provisioning or repeated "recognition-avoidance" coevolution where growth-enhancing paternally derived alleles evolve to escape recognition by maternal genes targeted to suppress their effect. Here, we develop predictions to distinguish between these two mechanisms when considering crosses among populations that have reached different equilibria in this intersexual arms race. We test these predictions using crosses within and among populations of Dalechampia scandens (Euphorbiaceae) that presumably have experienced different intensities of intersexual conflict, as inferred from their historical differences in mating system. In crosses where the paternal population was more outcrossed than the maternal population, hybrid seeds were larger than those normally produced in the maternal population, whereas when the maternal population was more outcrossed, hybrid seeds were smaller than normal. These results confirm the importance of mating systems in determining the intensity of intersexual conflict over maternal investment and provide strong support for a tug-of-war mechanism operating in this conflict. They also yield clear predictions for the fitness consequences of gene flow among populations with different mating histories.

Reversible colour change in leaves enhances pollinator attraction and reproductive success in Saururus chinensis (Saururaceae).

Background and Aims: Although there has been much experimental work on leaf colour change associated with selection generated by abiotic environmental factors and antagonists, the role of leaf colour change in pollinator attraction has been largely ignored. We tested whether whitening of the apical leaves subtending the inflorescences of Saururus chinensis during flowering enhances pollinator attraction, and whether re-greening of the white leaves after flowering increases carbon assimilation and promotes seed development. Methods: White leaves were removed or covered, and the effects of these manipulations on pollinator visitation and subsequent reproductive success were assessed. The net photosynthetic rates of leaves of different colour were measured and their photosynthetic contributions to seed development were evaluated. Key Results: Saururus chinensis is able to self-pollinate autonomously, but depends largely on flies for pollination. White leaves had different reflectance spectra from green leaves, and white leaves attracted significantly more pollinators and led to significantly higher fruit and seed set. Although leaf whitening resulted in a reduction in photosynthetic capacity, it translated into only a small decrease in seed mass. When leaves had turned back from white to green after flowering their photosynthetic capacity was similar to that of 'normal' green leaves and promoted seed development. Conclusions: The reversible leaf colour change in S. chinensis appears to be adaptive because it enhances pollination success during flowering, with a small photosynthetic cost, while re-greening of these leaves after flowering helps to meet the carbon requirements for seed development.