A sensory bias overrides learned preferences of bumblebees for honest signals in Mimulus guttatus.

Insect pollinators readily learn olfactory cues, and this is expected to select for 'honest signals' that provide reliable information about floral rewards. However, plants might alternatively produce signals that exploit pollinators' sensory biases, thereby relaxing selection for signal honesty. We examined the innate and learned preferences of Bombus impatiens for Mimulus guttatus floral scent phenotypes corresponding to different levels of pollen rewards in the presence and absence of the innately attractive floral volatile compound ß-trans-bergamotene. Bees learned to prefer honest signals after foraging on live M. guttatus flowers, but only exhibited this preference when presented floral scent phenotypes that did not include ß-trans-bergamotene. Our results suggest that a sensory bias for ß-trans-bergamotene overrides the ability of B. impatiens to use honest signals when foraging on M. guttatus. This may represent a deceptive pollination strategy that allows plants to minimize investment in costly rewards without incurring reduced rates of pollinator visitation.

Decline of six native mason bee species following the arrival of an exotic congener.

A potential driver of pollinator declines that has been hypothesized but seldom documented is the introduction of exotic pollinator species. International trade often involves movement of many insect pollinators, especially bees, beyond their natural range. For agricultural purposes or by inadvertent cargo shipment, bee species successfully establishing in new ranges could compete with native bees for food and nesting resources. In the Mid-Atlantic United States, two Asian species of mason bee (Osmia taurus and O. cornifrons) have become recently established. Using pan-trap records from the Mid-Atlantic US, we examined catch abundance of two exotic and six native Osmia species over the span of fifteen years (2003-2017) to estimate abundance changes. All native species showed substantial annual declines, resulting in cumulative catch losses ranging 76-91% since 2003. Exotic species fared much better, with O. cornifrons stable and O. taurus increasing by 800% since 2003. We characterize the areas of niche overlap that may lead to competition between native and exotic species of Osmia, and we discuss how disease spillover and enemy release in this system may result in the patterns we document.

Interactions between artificial light at night, soil moisture, and plant density affect the growth of a perennial wildflower.

Artificial light at night (ALAN) has been shown to alter aspects of plant growth, but we are not aware of any studies that have examined whether the effects of ALAN on plants depend upon the backdrop of variation in other abiotic factors that plants encounter in field populations. We conducted a field experiment to investigate whether ALAN affects the growth and anti-herbivore defenses of common milkweed, Asclepias syriaca, and whether the effects of ALAN are influenced by plant density or soil moisture content. Artificial light at night, soil moisture, and plant density were manipulated according to a split-plot factorial design. Although increasing soil moisture by watering had no significant effects on latex exudation, attributes of plant growth generally responded positively to watering. The basal stem diameter (BSD) and height of plants were affected by ALAN × soil moisture interactions. For both of these variables, the positive effects of ALAN were greater for plants that were not watered than for plants that were. Basal stem diameter was also affected by an ALAN × plant density interaction, and the positive effect of ALAN on BSD was greater in the low-density treatment than in the high-density treatment. Our results demonstrate that the effects of ALAN on plant growth can be altered by soil moisture and plant density. Consequently, the effects of ALAN on plants in nature may not be consistent with existing frameworks that do not account for critical abiotic variables such as water availability or biotic interactions between plants such as competition.

Exotic tree and shrub invasions alter leaf-litter microflora and arthropod communities.

Approximately 90% of all annual net primary productivity in temperate deciduous forests ends up entering the detritus food web as leaf litter. Due to chemical and physical differences from native litter, inputs from invasive species may impact the litter-dwelling community and ecosystem processes. We compared leaf-litter nutritional quality and decomposition rates from two invasive shrubs, Lonicera maackii and Rhamnus davurica, and the invasive tree Ailanthus altissima to litter from native oak-hickory forest in the Shenandoah Valley of Virginia, USA. We sampled litter from both invaded and uninvaded habitats and conducted litter colonization experiments to test for effects on microflora and the litter-dwelling arthropod communities. Litter from all three invasive species decomposed more rapidly than native litter, with native habitats averaging two to nearly five times as much litter by June. Invasive litter had higher nitrogen concentration and lower C:N ratios than native litter. Invasive litter supported greater growth of bacteria and fungi. Higher numbers of arthropods colonized invasive litter than native litter, but litter arthropod numbers on the forest floor of invaded habitats dropped in the early summer as litter decomposed. Litter had no effect on arthropod richness. Over short time scales, our results indicate that these invasive species represent beneficial, novel resources for the litter-dwelling community. However, the short-lived nature of this resource resulted in a crash in the abundance of the litter-dwelling organisms once the litter decomposed. As a whole, native habitat seems to support a larger, more stable litter-dwelling community over the course of a growing season.

The effect of nitrogen availability and water conditions on competition between a facultative CAM plant and an invasive grass.

Plants with crassulacean acid metabolism (CAM) are increasing their abundance in drylands worldwide. The drivers and mechanisms underlying the increased dominance of CAM plants and CAM expression (i.e., nocturnal carboxylation) in facultative CAM plants, however, remain poorly understood. We investigated how nutrient and water availability affected competition between Mesembryanthemum crystallinum (a model facultative CAM species) and the invasive C3 grass Bromus mollis that co-occur in California's coastal grasslands. Specifically we investigated the extent to which water stress, nutrients, and competition affect nocturnal carboxylation in M. crystallinum. High nutrient and low water conditions favored M. crystallinum over B. mollis, in contrast to high water conditions. While low water conditions induced nocturnal carboxylation in 9-week-old individuals of M. crystallinum, in these low water treatments, a 66% reduction in nutrient applied over the entire experiment did not further enhance nocturnal carboxylation. In high water conditions M. crystallinum both alone and in association with B. mollis did not perform nocturnal carboxylation, regardless of the nutrient levels. Thus, nocturnal carboxylation in M. crystallinum was restricted by strong competition with B. mollis in high water conditions. This study provides empirical evidence of the competitive advantage of facultative CAM plants over grasses in drought conditions and of the restricted ability of M. crystallinum to use their photosynthetic plasticity (i.e., ability to switch to CAM behavior) to compete with grasses in well-watered conditions. We suggest that a high drought tolerance could explain the increased dominance of facultative CAM plants in a future environment with increased drought and nitrogen deposition, while the potential of facultative CAM plants such as M. crystallinum to expand to wet environments is expected to be limited.

Effects of soil nitrogen availability and native grass diversity on exotic forb dominance.

Exotic plants are often most successful in high resource environments. By drawing down available resources, species-rich communities may be able to reduce exotic success when resource supply is elevated. We tested the prediction that exotic success would be greatest in species-poor communities when nitrogen availability is high. We also tested two underlying assumptions of this prediction: species-rich communities draw down soil nitrogen availability more than species-poor communities following fertilization and exotic success increases when soil nitrogen availability is high. In a restored grassland where native grass diversity was manipulated (one, three, or five-species) seven years earlier to form a gradient in species richness, we manipulated nitrogen availability directly via fertilization, and indirectly via burning. We then examined the success of the exotic forb Galium verum L. Contrary to our prediction, diversity and nutrient treatments did not jointly influence exotic success. Instead, one-time fertilization increased exotic biomass in the first year of the study. This likely occurred because the effect of nutrient treatments on nitrogen availability was independent of diversity treatment. Thus, we found no evidence that species-rich communities are better able to reduce exotic biomass when nitrogen is added than are species-poor communities. This suggests that in some systems, the effects of increasing species richness can be overwhelmed by the effects of nutrient addition that promote exotic success.

The effects of leaf litter nutrient pulses on Alliaria petiolata performance.

Nutrient pulses can facilitate species establishment and spread in new habitats, particularly when one species more effectively uses that nutrient pulse. Biological differences in nutrient acquisition between native and exotic species may facilitate invasions into a variety of habitats including deciduous forest understories. Alliaria petiolata (Bieb.) Cavara & Grande is an important invader of deciduous forest understories throughout much of North America. These understory communities contain many species which perform the majority of their growth and reproduction before canopy closure in spring. Because A. petiolata is a wintergreen biennial that can be active during autumn and winter, it may utilize nutrients released from decaying leaf litter before its competitors. To investigate this we manipulated the timing of leaf litter addition (fall or spring) and experimentally simulated the nutrient pulse from decaying leaves using artificial fertilizer. To determine whether A. petiolata affected the abundance of understory competitors, we also removed A. petiolata from one treatment. A. petiolata that received early nutrients exhibited greater growth. Treatments receiving fall leaf litter or artificial nutrients had greater A. petiolata adult biomass than plots receiving spring nutrient additions (leaf litter or artificial nutrients). However, fall leaf litter addition had no effect on the richness of competitor species. Thus, wintergreen phenology may contribute to the spread of A. petiolata through deciduous forest understories, but may not explain community-level impacts of A. petiolata in deciduous forests.

Lack of Melanopsin Is Associated with Extreme Weight Loss in Mice upon Dietary Challenge.

Metabolic disorders have been established as major risk factors for ocular complications and poor vision. However, little is known about the inverse possibility that ocular disease may cause metabolic dysfunction. To test this hypothesis, we assessed the metabolic consequences of a robust dietary challenge in several mouse models suffering from retinal mutations. To this end, mice null for melanopsin (Opn4-/-), the photopigment of intrinsically photosensitive retinal ganglion cells (ipRGCs), were subjected to five weeks of a ketogenic diet. These mice lost significantly more weight than wild-type controls or mice lacking rod and cone photoreceptors (Pde6brd1/rd1). Although ipRGCs are critical for proper circadian entrainment, and circadian misalignment has been implicated in metabolic pathology, we observed no differences in entrainment between Opn4-/- and control mice. Additionally, we observed no differences in any tested metabolic parameter between these mouse strains. Further studies are required to establish the mechanism giving rise to this dramatic phenotype observed in melanopsin-null mice. We conclude that the causality between ocular disease and metabolic disorders merits further investigation due to the popularity of diets that rely on the induction of a ketogenic state. Our study is a first step toward understanding retinal pathology as a potential cause of metabolic dysfunction.

Variation in reward quality and pollinator attraction: the consumer does not always get it right.

Nearly all bees rely on pollen as the sole protein source for the development of their larvae. The central importance of pollen for the bee life cycle should exert strong selection on their ability to locate the most rewarding sources of pollen. Despite this importance, very few studies have examined the influence of intraspecific variation in pollen rewards on the foraging decisions of bees. Previous studies have demonstrated that inbreeding reduces viability and hence protein content in Mimulus guttatus (seep monkeyflower) pollen and that bees strongly discriminate against inbred in favour of outbred plants. We examined whether variation in pollen viability could explain this preference using a series of choice tests with living plants, artificial plants and olfactometer tests using the bumble bee Bombus impatiens. We found that B. impatiens preferred to visit artificial plants provisioned with fertile anthers over those provisioned with sterile anthers. They also preferred fertile anthers when provided only olfactory cues. These bumble bees were unable to discriminate among live plants from subpopulations differing dramatically in pollen viability, however. They preferred outbred plants even when those plants were from subpopulations with pollen viability as low as the inbred populations. Their preference for outbred plants was evident even when only olfactory cues were available. Our data showed that bumble bees are able to differentiate between anthers that provide higher rewards when cues are isolated from the rest of the flower. When confronted with cues from the entire flower, their choices are independent of the quality of the pollen reward, suggesting that they are responding more strongly to cues unassociated with rewards than to those correlated with rewards. If so, this suggests that a sensory bias or some level of deception may be involved with advertisement to pollinators in M. guttatus.

Inbreeding in Mimulus guttatus reduces visitation by bumble bee pollinators.

Inbreeding in plants typically reduces individual fitness but may also alter ecological interactions. This study examined the effect of inbreeding in the mixed-mating annual Mimulus guttatus on visitation by pollinators (Bombus impatiens) in greenhouse experiments. Previous studies of M. guttatus have shown that inbreeding reduced corolla size, flower number, and pollen quantity and quality. Using controlled crosses, we produced inbred and outbred families from three different M. guttatus populations. We recorded the plant genotypes that bees visited and the number of flowers probed per visit. In our first experiment, bees were 31% more likely to visit outbred plants than those selfed for one generation and 43% more likely to visit outbred plants than those selfed for two generations. Inbreeding had only a small effect on the number of flowers probed once bees arrived at a genotype. These differences were explained partially by differences in mean floral display and mean flower size, but even when these variables were controlled statistically, the effect of inbreeding remained large and significant. In a second experiment we quantified pollen viability from inbred and self plants. Bees were 37-54% more likely to visit outbred plants, depending on the population, even when controlling for floral display size. Pollen viability proved to be as important as floral display in predicting pollinator visitation in one population, but the overall explanatory power of a multiple regression model was weak. Our data suggested that bees use cues in addition to display size, flower size, and pollen reward quality in their discrimination of inbred plants. Discrimination against inbred plants could have effects on plant fitness and thereby reinforce selection for outcrossing. Inbreeding in plant populations could also reduce resource quality for pollinators, potentially resulting in negative effects on pollinator populations.

Interactions between insect herbivores and plant mating systems.

Self-pollination is common in plants, and limited seed and pollen dispersal can create localized inbreeding even within outcrossing plants. Consequently, insects regularly encounter inbred plants in nature. Because inbreeding results in elevated homozygosity, greater expression of recessive alleles, and subsequent phenotypic changes in inbred plants, inbreeding may alter plant-insect interactions. Recent research has found that plant inbreeding alters resistance and tolerance to herbivores, alters the attraction and susceptibility of plants to insects that vector plant pathogens, and alters visitation rates of insect pollinators. These results suggest that interactions with insects can increase or decrease inbreeding depression (the loss of fitness due to self-fertilization) and subsequently alter the evolution of selfing within plant populations. Future work needs to focus on the mechanisms underlying genetic variation in the effects of inbreeding on plant-insect interactions and the consequences of altered plant-insect interactions on the evolution of plant defense and plant mating systems.

A multidimensional approach to understanding floral function and form.

PREMISE OF THE STUDY: Variation in plant mating systems is a fundamental component of the diversity of floral form and function. Mating systems have a strong influence on the distribution and expression of genetic variation, which in turn can influence the course of mating system evolution. It has long been appreciated, however, that ecological interactions may provide much of the impetus behind these evolutionary changes. • METHODS: This commentary reviews the Special Section in the American Journal of Botany (vol. 100, issue 6) that features novel research on the role of interactions between plants and their pollinators, seed dispersers, herbivores, and interspecific competitors in the evolution of selfing rates and gender. These studies vary in approach from empirical experiments, to phylogenetic comparisons, to theoretical models, to literature reviews as they each attempt to shed new light on longstanding questions about the selective forces and evolutionary pathways that have led to the diversified means by which plants promote or discourage self-fertilization. • KEY RESULTS: Evidence is provided that indicates that ecological interactions (e.g., herbivory and heterospecific pollen deposition by pollinators) can strongly influence the relative advantages of selfing and outcrossing. Ecological interactions can also influence allocation to sexual functions, which will influence individual and population outcrossing rates and possibly the evolutionary path of gender expression. • CONCLUSIONS: Mating systems evolution is clearly a multidimensional problem. Research that places the study of mating and sexual systems within a more realistic ecological context will no doubt reveal more complexity as we move toward a better understanding of plant diversity.

Tests for the joint evolution of mating system and drought escape in Mimulus.

BACKGROUND AND AIMS: Self-fertilizing taxa are often found at the range margins of their progenitors, where sub-optimal habitats may select for alternative physiological strategies. The extent to which self-fertilization is favoured directly vs. arising indirectly through correlations with other adaptive life history traits is unclear. Trait responses to selection depend on genetic variation and covariation, as well as phenotypic and genetic responses to altered environmental conditions. We tested predictions of the hypothesis that self-fertilization in Mimulus arises through direct selection on physiological and developmental traits that allow seasonal drought escape. METHODS: Phenotypic selection on mating system and drought escape traits was estimated in field populations of M. guttatus. In addition, trait phenotype and phenotypic selection were compared between experimental wet and dry soil in two greenhouse populations each of M. guttatus and M. nasutus. Finally, genetic variation and covariation for traits were compared between wet and dry soil treatments in a greenhouse population of M. guttatus. KEY RESULTS: Consistent with predictions, selection for early flowering was generally stronger than for mating system traits, and selection for early flowering was stronger in dry soil. Inconsistent with predictions, selection for water-use efficiency was largely absent; selection for large flowers was stronger than for drought escape in the field; and most drought escape and mating system traits were not genetically correlated. A positive genetic correlation between flowering time and flower size, which opposed the adaptive contour, emerged only in wet soil, suggesting that variation in water availability may maintain variation in these traits. Plastic responses to soil moisture treatments supported the idea that taxonomic divergence could have been facilitated by plasticity in flowering time and selfing. CONCLUSIONS: The hypothesis that plant mating systems may evolve indirectly via selection on correlated life history characteristics is plausible and warrants increased attention.

On quantifying tolerance of herbivory for comparative analyses.

As the evolutionary importance of plant tolerance of herbivory is increasingly appreciated, more and more studies are not just measuring a plant's tolerance, but are comparing tolerance among plant genotypes, populations, species, and environments. Here, we suggest that caution must be taken in such comparative studies in the choice of measurement scales (and data transformations) for damage levels and plant performance. We demonstrate with a simple scenario of two plant groups of equal tolerance how the choice of scales can lead one to infer that the first group is more tolerant, the second group is more tolerant, or the two groups are equally tolerant-using the identical dataset. We conclude that to make reliable, logically consistent inferences when comparing tolerances among groups of plants, damage and performance should both be on an additive scale or both on a multiplicative scale.

Effects of herbivory and inbreeding on the pollinators and mating system of Mimulus guttatus (Phrymaceae).

Most models of mating system evolution predict mixed mating to be unstable, although it is commonly reported from nature. Ecological interactions with mutualistic pollinators can help account for this discrepancy, but antagonists such as herbivores are also likely to play a role. In addition, inbreeding can alter ecological interactions and directly affect selfing rates, which may also contribute to maintaining mating system variation. We explored herbivore and inbreeding effects on pollinator behavior and selfing rates in Mimulus guttatus. First, individual spittlebug (Philaenus spumarius) herbivores were applied to native plants in two populations. Spittlebugs reduced flower size, increased anther-stigma distance, and increased selfing rates. A second experiment factorially crossed spittlebug treatment with inbreeding history (self- vs. cross-fertilized), using potted plants in arrays. Spittlebugs did not affect pollinator behavior, but they reduced flower size and nearly doubled the selfing rate. Inbreeding reduced the frequency of pollinator visits and increased flower-handling time, and this may be the first report that inbreeding affects pollinator behavior. Selfing rates of inbred plants were reduced by one half, which may reflect early inbreeding depression or altered pollinator behavior. The contrasting effects of herbivory and inbreeding on selfing rates may help maintain mating system variation in M. guttatus.

Inbreeding depression and selfing rate of Ipomoea hederacea var. integriuscula (Convolvulaceae).

Inbreeding depression and selfing rate were investigated in the self-compatible vine Ipomoea hederacea to assess the variability of the breeding system. Inbreeding depression differed between populations and the magnitude varied at germination, growth (as measured by aboveground biomass), and reproductive potential. Plants from Macon County, Alabama, USA, had significant inbreeding depression (31%) at germination, but no significant inbreeding depression for aboveground biomass or number of reproductive structures (buds and flowers) at 45 d post germination in the greenhouse or in the field. Plants from Morgan County, Alabama, however, had significant inbreeding depression (>50%) for all three stages in the greenhouse. In allozyme comparisons, five of the 11 I. hederacea populations surveyed had high selfing rates (66.66-92.53%) and high levels of homozygosity (F(IS) = 0.500-0.861) in 2003, and three of four populations surveyed in 2004 had selfing rates that exceeded 50%. High selfing rates, high levels of homozygosity, and low levels of inbreeding depression suggest that inbreeding depression may not present a significant barrier to the transmission of selfing alleles in some populations of I. hederacea, but does not account for the maintenance of a mixed mating system in other populations.

Recent approaches into the genetic basis of inbreeding depression in plants.

Predictions for the evolution of mating systems and genetic load vary, depending on the genetic basis of inbreeding depression (dominance versus overdominance, epistasis and the relative frequencies of genes of large and small effect). A distinction between the dominance and overdominance hypotheses is that deleterious recessive mutations should be purged in inbreeding populations. Comparative studies of populations differing in their level of inbreeding and experimental approaches that allow selection among inbred lines support this prediction. More direct biometric approaches provide strong support for the importance of partly recessive deleterious alleles. Investigators using molecular markers to study quantitative trait loci (QTL) often find support for overdominance, though pseudo-overdominance (deleterious alleles linked in repulsion) may bias this perception. QTL and biometric studies of inbred lines often find evidence for epistasis, which may also contribute to the perception of overdominance, though this may be because of the divergent lines initially crossed in QTL studies. Studies of marker segregation distortion commonly uncover genes of major effect on viability, but these have only minor contributions to inbreeding depression. Although considerable progress has been made in understanding the genetic basis of inbreeding depression, we feel that all three aspects merit more study in natural plant populations.

Inbreeding alters resistance to insect herbivory and host plant quality in Mimulus guttatus (Scrophulariaceae).

Previous studies have demonstrated genetic variation for resistance to insect herbivores and host plant quality. The effect of plant mating system, an important determinant of the distribution of genetic variation, on host plant characteristics has received almost no attention. This study used a controlled greenhouse experiment to examine the effect of self- and cross-pollination in Mimulus guttatus (Scrophulariaceae) on resistance to and host plant quality for the xylem-feeding spittlebug Philaenus spumarius (Homoptera: Cercopidae). Spittlebugs were found to have a negative effect on two important fitness components in M. guttatus, flower production and above ground biomass. One of two M. guttatus populations examined showed a significant interaction between the pollination and herbivore treatments. In this case, the detrimental effects of herbivores on biomass and flower production were much more pronounced in inbred (self) plants. The presence of spittlebug nymphs increased inbreeding depression by as much as three times. Pollination treatments also had significant effects on important components of herbivore fitness, but these effects were in opposite directions in our two host plant populations. Spittlebug nymphs maturing on self plants emerged as significantly larger adults in one of our host plant populations, indicating that inbreeding increased host plant quality. In our second host plant population, spittlebug nymphs took significantly longer to develop to adulthood on self plants, indicating that inbreeding decreased host plant quality. Taken together these results suggest that the degree of inbreeding in host plant populations can have important and perhaps complex effects on the dynamics of plant-herbivore interactions and on mating-system evolution in the host.

THE EFFECTS OF FIVE GENERATIONS OF ENFORCED SELFING ON POTENTIAL MALE AND FEMALE FUNCTION IN MIMULUS GUTTATUS.

In prior work we detected no significant inbreeding depression for pollen and ovule production in the highly selfing Mimulus micranthus, but both characters showed high inbreeding depression in the mixed-mating M. guttatus. The goal of this study was to determine if the genetic load for these traits in M. guttatus could be purged in a program of enforced selfing. These characters should have been under much stronger selection in our artificial breeding program than previously reported characters such as biomass and total flower production because, for example, plants unable to produce viable pollen could not contribute to future generations. Purging of genetic load was investigated at the level of both the population and the individual maternal line within two populations of M. guttatus. Mean ovule number, pollen number, and pollen viability declined significantly as plants became more inbred. The mean performance of outcross progeny generated from crosses between pairs of maternal inbred lines always exceeded that of self progeny and was fairly constant for each trait through all five generations. The consistent performance of outcross progeny and the universally negative relationships between performance and degree of inbreeding are interpreted as evidence for the weakness of selection relative to the quick fixation of deleterious alleles due to drift during the inbreeding process. The selective removal (purging) of deleterious alleles from our population would have been revealed by an increase in performance of outcross progeny or an attenuation of the effects of increasing homozygosity. The relationships between the mean of each of these traits and the expected inbreeding coefficient were linear, but one population displayed a significant negative curvilinear relationship between the log of male fertility (a function of pollen number and viability) and the inbreeding coefficient. The generally linear form of the responses to inbreeding were taken as evidence consistent with an additive model of gene action, but the negative curvilinear relationship between male fertility and the inbreeding coefficient suggested reinforcing epistasis. Within both populations there was significant genetic variation among maternal lineages for the response to inbreeding in all traits. Although all inbred lineages declined at least somewhat in performance, several maternal lines maintained levels of performance just below outcross means even after four or five generations of selfing. We suggest that selection among maternal lines will have a greater effect than selecting within lines in lowering the genetic load of populations.

THE RELATIONSHIP BETWEEN MATING-SYSTEM CHARACTERS AND INBREEDING DEPRESSION IN MIMULUS GUTTATUS.

Several recent theoretical considerations of mating-system evolution predict within-population covariation between levels of inbreeding depression and genetically controlled mating-system characters. If inbreeding depression is caused by deleterious recessive alleles, families with characters that promote self-pollination should show lower levels of inbreeding depression than families with characters that promote outcrossing. The converse is expected if inbreeding depression is due to overdominant allelic interactions. Whether these associations between mating-system and viability loci evolve will have important consequences for mating-system evolution. The evolution of selfing within the genus Mimulus is associated with a reduction in stigma-anther separation (i.e., a loss of herkogamy) and high autogamous seed set. In this study we compared families from two M. guttatus populations that differed genetically in their degree of stigma-anther separation. In one of these populations we also compared families that differed genetically in the degree to which they autogamously set seed in a pollinator-free greenhouse. Dams often differed significantly in levels of inbreeding depression for aboveground biomass and flower production, but variation in inbreeding depression was never explained by herkogamy class or autogamy class. Several factors might account for why families with traits associated with selfing did not show lower inbreeding depression, and these are discussed. Our study also demonstrated significant variation among self progeny from a given female likely due to differences in pollination date and position of fruit maturation. The detection of significant dam × sire interactions suggests biparental inbreeding or differences in combining ability for specific pairs of parents.