A Comparative Approach to Understanding Floral Adaptation to Climate and Pollinators During Diversification in European and Mediterranean Silene.

Pollinator selection on floral traits is a well-studied phenomenon, but less is known about the influence of climate on this species interaction. Floral trait evolution could be a result of both adaptation to climate and pollinator-mediated selection. In addition, climate may also determine pollinator communities, leading to an indirect influence of climate on floral traits. In this study, we present evidence of both direct and indirect effects of climate on plant morphology through a phylogenetic comparative analysis of the relationships between climate, pollinators, and morphology in 89 European and Mediterranean Silene species. Climate directly influences vegetative morphology, where both leaf size and internode length were found to be smaller in habitats that are warmer in the driest quarter of the year and that have more precipitation in the coldest quarter of the year. Similarly, flower size was directly influenced by climate, where smaller calyxes were also associated with habitats that are warmer in the driest quarter of the year. These results suggest that reduced leaf and flower size promote water conservation in species that occupy arid climates. Floral traits also evolved in response to pollinators, with elongated calyxes associated with nocturnal pollination, though we also found evidence that climate influences pollinator distribution. Nocturnal pollinators of Silene are found in habitats that have more temperature evenness across seasons than diurnal pollinators. Correspondingly, nocturnally-pollinated Silene are more likely to occur in habitats that have lower daily temperature fluctuation and more temperature evenness across seasons. Altogether these results show that climate can directly influence vegetative and floral morphology, but it can also affect pollinator distribution, which in turn drives floral adaptation. Our study therefore suggests that climate mediates the influence of species interactions on trait evolution by imposing direct selective demands on floral phenotypes and by determining the pollinator community that imposes its own selective demands on flowers.

Rapid reversal of a potentially constraining genetic covariance between leaf and flower traits in Silene latifolia.

Genetic covariance between two traits generates correlated responses to selection, and may either enhance or constrain adaptation. Silene latifolia exhibits potentially constraining genetic covariance between specific leaf area (SLA) and flower number in males. Flower number is likely to increase via fecundity selection but the correlated increase in SLA increases mortality, and SLA is under selection to decrease in dry habitats. We selected on trait combinations in two selection lines for four generations to test whether genetic covariance could be reduced without significantly altering trait means. In one selection line, the genetic covariance changed sign and eigenstructure changed significantly, while in the other selection line eigenstructure remained similar to the control line. Changes in genetic variance-covariance structure are therefore possible without the introduction of new alleles, and the responses we observed suggest that founder effects and changes in frequency of alleles of major effect may be acting to produce the changes.

The defensive benefit and flower number cost of selenium accumulation in Brassica juncea.

Some plant species accumulate selenium in their tissues in quantities far above soil concentrations, and experiments demonstrate that selenium can serve as a defence against herbivores and pathogens. However, selenium may also cause oxidative stress and reduce growth in plants. We measured growth, selenium accumulation and herbivory in four varieties of the selenium accumulator Brassica juncea to investigate the cost of accumulation as well as its benefit in reducing herbivory. We measured selenium levels, plant size and flower number in four varieties of B. juncea watered with sodium selenate or treated as controls. We also conducted no-choice herbivory trials on leaves from both treatments with the specialist herbivore Pieris rapae. The selenate treatment slightly increased leaf number over the control, but tissue concentrations of selenium and flower number were negatively correlated in some varieties. In herbivory trials, leaves from the plants in the selenate treatment lost less leaf tissue, and the majority of larvae given leaves from selenate-treated plants ate very little leaf tissue at all. In the variety with the highest selenium accumulation, leaves from selenate-treated plants that showed reduced flower production also experienced less herbivory in feeding trials. The protective advantage of greater selenium accumulation may be offset by negative effects on reproduction, and the relatively low level of selenium accumulation in this species as compared to more extreme hyperaccumulators could reflect the minimum level necessary to enhance protection from herbivory.

Spatiotemporal variation in deer browse and tolerance in a woodland herb.

Herbivory can shape the dynamics of plant populations, including effects on survival and reproduction, and is in turn affected by environmental factors that vary in space and time. White-tailed deer are significant herbivores in North America that have been broadly documented to affect plant reproductive success. If variation in the frequency and impact of herbivory by deer correlates with a broad-scale latitudinal gradient, climactic effects may be important for shaping plant-herbivore interactions, Alternatively, a lack of broad-scale gradients would suggest local factors such as plant community composition and deer densities are affecting herbivory. To investigate broad-scale patterns of deer herbivory, we examined the frequency and reproductive consequences of deer browse over three years in 17 populations of Campanulastrum americanum spanning the latitudinal extent of its range. Even though deer are overabundant throughout the range of C. americanum, we found spatiotemporal variation in deer browse frequency (0-0.96, mean 0.46) and its effects on plant reproductive success. The four southernmost populations experienced high levels of herbivory, and were responsible for generating a negative relationship between latitude and herbivory. In general, patterns of variation in the frequency and impact of herbivory across the entire latitudinal gradient pointed to the importance of local rather than broad-scale factors. Within a population, deer consumed larger plants. Across many populations and years, average fitnesses of browsed and uneaten plants were similar, suggesting that plants are tolerant to browse. However, since large plants have greater reproductive success and are more likely to be browsed, tolerance may be influenced by plant size. When plant size was accounted for, most populations did not fully compensate for browsing. There was no relationship between browsing intensity and tolerance, suggesting that browsing may be too variable to consistently select for tolerance, or that increases in deer density are too recent for increased tolerance to evolve.

Very-high-resolution time-lapse photography for plant and ecosystems research.

PREMISE OF THE STUDY: Traditional photography is a compromise between image detail and area covered. We report a new method for creating time-lapse sequences of very-high-resolution photographs to produce zoomable images that facilitate observation across a range of spatial and temporal scales. • METHODS AND RESULTS: A robotic camera mount and software were used to capture images of the growth and movement in Brassica rapa every 15 s in the laboratory. The resultant time-lapse sequence (http://timemachine.gigapan.org/wiki/Plant_Growth) captures growth detail such as circumnutation. A modified, solar-powered system was deployed at a remote field site in southern Arizona. Images were collected every 2 h over a 3-mo period to capture the response of vegetation to monsoon season rainfall (http://timemachine.gigapan.org/wiki/Arizona_Grasslands). • CONCLUSIONS: A technique for observing time sequences of both individual plant and ecosystem response at a range of spatial scales is available for use in the laboratory and in the field.

Elimination of a genetic correlation between the sexes via artificial correlational selection.

Genetic correlations between the sexes can constrain the evolution of sexual dimorphism and be difficult to alter, because traits common to both sexes share the same genetic underpinnings. We tested whether artificial correlational selection favoring specific combinations of male and female traits within families could change the strength of a very high between-sex genetic correlation for flower size in the dioecious plant Silene latifolia. This novel selection dramatically reduced the correlation in two of three selection lines in fewer than five generations. Subsequent selection only on females in a line characterized by a lower between-sex genetic correlation led to a significantly lower correlated response in males, confirming the potential evolutionary impact of the reduced correlation. Although between-sex genetic correlations can potentially constrain the evolution of sexual dimorphism, our findings reveal that these constraints come not from a simple conflict between an inflexible genetic architecture and a pattern of selection working in opposition to it, but rather a complex relationship between a changeable correlation and a form of selection that promotes it. In other words, the form of selection on males and females that leads to sexual dimorphism may also promote the genetic phenomenon that limits sexual dimorphism.

Environment-dependent intralocus sexual conflict in a dioecious plant.

Intralocus sexual conflict is a form of conflict that does not involve direct interactions between males and females. It arises when selection on a shared trait with a common genetic basis differs between the sexes. Environmental factors, such as resource availability, may influence the expression and evolutionary outcome of such conflict. We quantified the genetic variance-covariance matrix, G, for both sexes of Silene latifolia for floral and leaf traits, as well as the between-sex matrix, B. We also quantified selection on the sexes via survival for 2 yr in four natural populations that varied in water availability. Environment-dependent intralocus sexual conflict exists for specific leaf area, a trait that is genetically correlated between the sexes. Males experienced significant negative selection, but only in populations with relatively limited water availability. Females experienced weakly positive or significant stabilizing selection on the same trait. Specific leaf area is genetically correlated with flower size and number, which are sexually dimorphic in this species. The extent of intralocus sexual conflict varied with the environment. Resolution of such conflict is likely to be confounded, given that specific leaf area is highly genetically integrated with other traits that are also divergent between the sexes.

Sexual dimorphism in the quantitative-genetic architecture of floral, leaf, and allocation traits in Silene latifolia.

Many species exhibit sexual dimorphism in a variety of characters, and the underlying genetic architecture of dimorphism potentially involves sex-specific differences in the additive-genetic variance-covariance matrix (G) of dimorphic traits. We investigated the quantitative-genetic structure of dimorphic traits in the dioecious plant Silene latifolia by estimating G (including within-sex matrices, G(m), G(f), and the between-sex variance-covariance matrix, B), and the phenotypic variance-covariance matrix (P) for seven traits. Flower number was the most sexually dimorphic trait, and was significantly genetically correlated with all traits within each sex. Negative genetic correlations between flower size and number suggested a genetic trade-off in investment, but positive environmental correlations between the same traits resulted in no physical evidence for a trade-off in the phenotype. Between-sex genetic covariances for homologous traits were always greater than 0 but smaller than 1, showing that some, but not all, of the variation in traits is caused by genes or alleles with sex-limited expression. Using common principal-components analysis (CPCA), a maximum-likelihood (ML) estimation approach, and element-by-element comparison to compare matrices, we found that G(m) and G(f) differed significantly in eigenstructure because of dissimilarity in covariances involving leaf traits, suggesting the presence of variation in sex-limited genes with pleiotropic effects and/or linkage between sex-limited loci. The sex-specific structure of G is expected to cause differences in the correlated responses to selection within each sex, promoting the further evolution and maintenance of dimorphism.

Investigating the independent evolution of the size of floral organs via G-matrix estimation and artificial selection.

The attractiveness of a plant to pollinators is dependent on both the number of flowers produced and the size of the petals. However, limiting resources often result in a size/number trade-off, whereby the plant can make either more flowers or larger flowers, but not both. If developmental genes underlying sepal and petal identity (some of which overlap) also influence size, then this shared genetic basis could constrain the independent evolution of floral size and attractiveness. Here, we determined whether the size of sepals and petals in the dioecious perennial, Silene latifolia, are developmentally independent by performing two experiments: a genetic variance-covariance experiment to estimate genetic correlations between calyx width, petal-limb length, flower mass, and number and a four-bout artificial-selection experiment to alter calyx width and estimate the correlated response in petal-limb length. In addition, we determined whether variation in petal-limb length is the result of cell expansion or cell proliferation. The first experiment revealed that petal-limb length is not genetically correlated with calyx width, and the second experiment confirmed this; selection on calyx width did not result in a predictable or significant change in petal-limb length. Flower number was negatively correlated with all the floral traits measured, indicating a flower size/number trade-off. Cell number, but not size, explained a significant amount of the variation in petal-limb length. We conclude that the size of the two outer floral organs can evolve independently. This species can therefore increase the number of flowers produced by decreasing investment in the calyx without simultaneously decreasing petal size and the attractiveness of each individual flower to pollinators.