Mimulus Laciniatus Research Articles

Fluctuating selection in a monkeyflower hybrid zone

Abstract While hybridization was viewed as a hindrance to adaptation and speciation by early evolutionary biologists, recent studies have demonstrated the importance of hybridization in facilitating evolutionary processes. However, it is still not well-known what role spatial and temporal variation in natural selection play in the maintenance of naturally occurring hybrid zones. To identify whether hybridization is adaptive between two closely related monkeyflower species, Mimulus guttatus and Mimulus laciniatus, we performed repeated reciprocal transplants between natural hybrid and pure species’ populations. We planted parental genotypes along with multiple experimental hybrid generations in a dry (2021) and extremely wet (2023) year in the Sierra Nevada, CA. By taking fine-scale environmental measurements, we found that the environment of the hybrid zone is more similar to M. laciniatus’s seasonally dry rocky outcrop habitat than M. guttatus’s moist meadows. In our transplants hybridization does not appear to be maintained by a consistent fitness advantage of hybrids over parental species in hybrid zones, but rather a lack of strong selection against hybrids. We also found higher fitness of the drought-adapted species, M. laciniatus, than M. guttatus in both species’ habitats, as well as phenotypic selection for M. laciniatus-like traits in the hybrid habitat in the dry year of our experiment. These findings suggest that in this system, hybridization might function to introduce drought-adapted traits and genes from M. laciniatus into M. guttatus, specifically in years with limited soil moisture. However, we also find evidence of genetic incompatibilities in second generation hybrids in the wetter year, which may balance a selective advantage of M. laciniatus introgression. Therefore, we find that hybridization in this system is both potentially adaptive and costly, and that the interaction of positive and negative selection likely determines patterns of gene flow between these Mimulus species.

Fluctuating selection in a Monkeyflower hybrid zone.

While hybridization was viewed as a hindrance to adaptation and speciation by early evolutionary biologists, recent studies have demonstrated the importance of hybridization in facilitating evolutionary processes. However, it is still not well-known what role spatial and temporal variation in natural selection play in the maintenance of naturally occurring hybrid zones. To identify whether hybridization is adaptive between two closely related monkeyflower species, Mimulus guttatus and Mimulus laciniatus, we performed repeated reciprocal transplants between natural hybrid and pure species' populations. We planted parental genotypes along with multiple experimental hybrid generations in a dry (2021) and extremely wet (2023) year in the Sierra Nevada, CA. By taking fine scale environmental measurements, we found that the environment of the hybrid zone is more similar to M. laciniatus's seasonally dry rocky outcrop habitat than M. guttatus's moist meadows. In our transplants hybridization does not appear to be maintained by a consistent fitness advantage of hybrids over parental species in hybrid zones, but rather a lack of strong selection against hybrids. We also found higher fitness of the drought adapted species, M. laciniatus, than M. guttatus in both species' habitats, as well as phenotypic selection for M. laciniatus-like traits in the hybrid habitat in the dry year of our experiment. These findings suggest that in this system hybridization might function to introduce drought-adapted traits and genes from M. laciniatus into M. guttatus, specifically in years with limited soil moisture. However, we also find evidence of genetic incompatibilities in second generation hybrids in the wetter year, which may balance a selective advantage of M. laciniatus introgression. Therefore, we find that hybridization in this system is both potentially adaptive and costly, and that the interaction of positive and negative selection likely determines patterns of gene flow between these Mimulus species.

Local adaptation to an altitudinal gradient: The interplay between mean phenotypic trait variation and phenotypic plasticity in Mimulus laciniatus

Organisms can adapt to environmental heterogeneity through two mechanisms: (1) expression of population genetic variation or (2) phenotypic plasticity. In this study we investigated whether patterns of variation in both trait means and phenotypic plasticity along elevational and latitudinal clines in a North American endemic plant, Mimulus laciniatus, were consistent with local adaptation. We grew inbred lines of M. laciniatus from across the species’ range in two common gardens varying in day length to measure mean and plastic trait expression in several traits previously shown to be involved in adaptation to M. laciniatus’s rocky outcrop microhabitat: flowering time, size-related traits, and leaf shape. We tested for phenotypic plasticity and GxE, examined correlations between the mean phenotype and plasticity, and tested for a relationship between trait variation and population elevation and latitude. We did not find a strong correlation between mean and plastic trait expression at the individual genotype level suggesting that they operate under independent genetic controls. We identified multiple traits that show patterns consistent with local adaptation to elevation: critical photoperiod, flowering time, flower size, mean leaf lobing, and leaf lobing plasticity. These trends occur along multiple geographically independent altitudinal clines indicating that selection is a more likely cause of this pattern than gene flow among nearby populations with similar trait values. We also found that population variation in mean leaf lobing is associated with latitude. Our results indicate that both having more highly lobed leaves and greater leaf shape plasticity may be adaptive at high elevation within M. laciniatus. Our data strongly suggest that traits known to be under divergent selection between M. laciniatus and close relative Mimulus guttatus are also under locally varying selection within M. laciniatus.

Populations Are Differentiated in Biological Rhythms without Explicit Elevational Clines in the Plant Mimulus laciniatus.

Environmental variation along an elevational gradient can yield phenotypic differentiation resulting from varying selection pressures on plant traits related to seasonal responses. Thus, genetic clines can evolve in a suite of traits, including the circadian clock, that drives daily cycling in varied traits and that shares its genetic background with adaptation to seasonality. We used populations of annual Mimulus laciniatus from different elevations in the Sierra Nevada in California to explore among-population differentiation in the circadian clock, flowering responses to photoperiod, and phenological traits (days to cotyledon emergence, days to flowering, and days to seed ripening) in controlled common-garden conditions. Further, we examined correlations of these traits with environmental variables related to temperature and precipitation. We observed that the circadian period in leaf movement was differentiated among populations sampled within about 100 km, with population means varying by 1.6 h. Significant local genetic variation occurred within 2 populations in which circadian period among families varied by up to 1.8 h. Replicated treatments with variable ecologically relevant photoperiods revealed marked population differentiation in critical day length for flowering that ranged from 11.0 to 14.1 h, corresponding to the time period between late February and mid-May in the wild. Flowering time varied among populations in a 14-h photoperiod. Regardless of this substantial population-level diversity, obvious linear clinality in trait variability across elevations could not be determined based on our genotypic sample; it is possible that more complex spatial patterns of variation arise in complex terrains such as those in the Sierra Nevada. Moreover, we did not find statistically significant bivariate correlations between population means of different traits. Our research contributes to the understanding of genetic variation in the circadian clock and in seasonal responses in natural populations, highlighting the need for more comprehensive investigations on the association between the clock and other adaptive traits in plants.

Evidence for adaptive responses to historic drought across a native plant species range.

As climatic conditions change, species will be forced to move or adapt to avoid extinction. Exacerbated by ongoing climate change, California recently experienced a severe and exceptional drought from 2011 to 2017. To investigate whether an adaptive response occurred during this event, we conducted a “resurrection” study of the cutleaf monkeyflower (Mimulus laciniatus), an annual plant, by comparing trait means and variances of ancestral seed collections (“pre‐drought”) with contemporary descendant collections (“drought”). Plants were grown under common conditions to test whether this geographically restricted species has the capacity to respond evolutionarily to climate stress across its range. We examined if traits shifted in response to the recent, severe drought and included populations across an elevation gradient, including populations at the low‐ and high‐elevation edges of the species range. We found that time to seedling emergence in the drought generation was significantly earlier than in the pre‐drought generation, a response consistent with drought adaptation. Additionally, trait variation in days to emergence was reduced in the drought generation, which suggests selection or bottleneck events. Days to first flower increased significantly by elevation, consistent with climate adaptation across the species range. Drought generation plants were larger and had greater reproduction, which was likely a carryover effect of earlier germination. These results demonstrate that rapid shifts in trait means and variances consistent with climate adaptation are occurring within populations, including peripheral populations at warm and cold climate limits, of a plant species with a relatively restricted range that has so far not shifted its elevation distribution during contemporary climate change. Thus, rapid evolution may mitigate, at least temporarily, range shifts under global climate change. This study highlights the need for better understanding rapid adaptation as a means for plant communities to cope with extraordinary climate events.

Differential adaptation to a harsh granite outcrop habitat between sympatric Mimulus species.

Understanding which environmental variables and traits underlie adaptation to harsh environments is difficult because many traits evolve simultaneously as populations or species diverge. Here, we investigate the ecological variables and traits that underlie Mimulus laciniatus' adaptation to granite outcrops compared to its sympatric, mesic-adapted progenitor, Mimulus guttatus. We use fine-scale measurements of soil moisture and herbivory to examine differences in selective forces between the species' habitats, and measure selection on flowering time, flower size, plant height, and leaf shape in a reciprocal transplant using M.laciniatus× M.guttatus F4 hybrids. We find that differences in drought and herbivory drive survival differences between habitats, that M.laciniatus and M.guttatus are each better adapted to their native habitat, and differential habitat selection on flowering time, plant stature, and leaf shape. Although early flowering time, small stature, and lobed leaf shape underlie plant fitness in M.laciniatus' seasonally dry environment, increased plant size is advantageous in a competitive mesic environment replete with herbivores like M.guttatus'. Given that we observed divergent selection between habitats in the direction of species differences, we conclude that adaptation to different microhabitats is an important component of reproductive isolation in this sympatric species pair.

The genetic architecture of local adaptation and reproductive isolation in sympatry within the Mimulus guttatus species complex.

The genetic architecture of local adaptation has been of central interest to evolutionary biologists since the modern synthesis. In addition to classic theory on the effect size of adaptive mutations by Fisher, Kimura and Orr, recent theory addresses the genetic architecture of local adaptation in the face of ongoing gene flow. This theory predicts that with substantial gene flow between populations local adaptation should proceed primarily through mutations of large effect or tightly linked clusters of smaller effect loci. In this study, we investigate the genetic architecture of divergence in flowering time, mating system-related traits, and leaf shape between Mimulus laciniatus and a sympatric population of its close relative M.guttatus. These three traits are probably involved in M.laciniatus' adaptation to a dry, exposed granite outcrop environment. Flowering time and mating system differences are also reproductive isolating barriers making them 'magic traits'. Phenotypic hybrids in this population provide evidence of recent gene flow. Using next-generation sequencing, we generate dense SNP markers across the genome and map quantitative trait loci (QTLs) involved in flowering time, flower size and leaf shape. We find that interspecific divergence in all three traits is due to few QTL of large effect including a highly pleiotropic QTL on chromosome 8. This QTL region contains the pleiotropic candidate gene TCP4 and is involved in ecologically important phenotypes in other Mimulus species. Our results are consistent with theory, indicating that local adaptation and reproductive isolation with gene flow should be due to few loci with large and pleiotropic effects.

Climate structures genetic variation across a species' elevation range: a test of range limits hypotheses

Gene flow may influence the formation of species range limits, and yet little is known about the patterns of gene flow with respect to environmental gradients or proximity to range limits. With rapid environmental change, it is especially important to understand patterns of gene flow to inform conservation efforts. Here we investigate the species range of the selfing, annual plant, Mimulus laciniatus, in the California Sierra Nevada. We assessed genetic variation, gene flow, and population abundance across the entire elevation-based climate range. Contrary to expectations, within-population plant density increased towards both climate limits. Mean genetic diversity of edge populations was equivalent to central populations; however, all edge populations exhibited less genetic diversity than neighbouring interior populations. Genetic differentiation was fairly consistent and moderate among all populations, and no directional signals of contemporary gene flow were detected between central and peripheral elevations. Elevation-driven gene flow (isolation by environment), but not isolation by distance, was found across the species range. These findings were the same towards high- and low-elevation range limits and were inconsistent with two common centre-edge hypotheses invoked for the formation of species range limits: (i) decreasing habitat quality and population size; (ii) swamping gene flow from large, central populations. This pattern demonstrates that climate, but not centre-edge dynamics, is an important range-wide factor structuring M.laciniatus populations. To our knowledge, this is the first empirical study to relate environmental patterns of gene flow to range limits hypotheses. Similar investigations across a wide variety of taxa and life histories are needed.

What can local and geographic population limits tell us about distributions?

Understanding the evolutionary and ecological factors that determine plant distributions is of primary importance in botanical research. These factors may vary in predictable ways across different spatial scales, and thus, we can leverage scale to reveal the underlying processes limiting plant distributions. We review various research considerations across local and geographic scales, including the investigation of dispersal and habitat limitation, evolutionary factors, abiotic and biotic factors, and research logistics. We also present two case studies, slender monkeyflower (Mimulus leptaleus) and cut-leaf monkeyflower (Mimulus laciniatus), in the California Sierra Nevada. At a local spatial scale (within 50 m), no seeds were produced from plants sown at sites located just beyond known patches of M. leptaleus, but within the species' geographic range. At a much broader spatial scale (kilometers), at the highest and lowest elevations of the species' range, we found greatly reduced abundance and fecundity in plants sown outside of the geographic range limits of M. laciniatus. These cases illustrate two contrasting spatial scales, yet agree in their illustration of strong habitat limitation. We end by discussing future avenues of research and by suggesting ways botanical researchers can frame their studies to maximize information gained on species requirements, distribution limits, and conservation among varying spatial scales.

Niche partitioning between close relatives suggests trade-offs between adaptation to local environments and competition.

Niche partitioning among close relatives may reflect trade-offs underlying species divergence and coexistence (e.g., between stress tolerance and competitive ability). We quantified the effects of habitat and congeneric species interactions on fitness for two closely related herbaceous plant species, Mimulus guttatus and Mimulus laciniatus, in three common habitat types within their sympatric range. Drought stress strongly reduced survival of M. guttatus in fast-drying seeps occupied by M. laciniatus, suggesting that divergent habitat adaptation maintains this niche boundary. However, neither seedling performance nor congeneric competition explained the absence of M. laciniatus from shady streams where M. guttatus thrives. M. laciniatus may be excluded from this habitat by competition with other species in the community or mature M. guttatus. Species performance and competitive ability were similar in sympatric meadows where plant community stature and the growing season length are intermediate between seeps and streams. Stochastic effects (e.g., dispersal among habitats or temporal variation) may contribute to coexistence in this habitat. Habitat adaptation, species interactions, and stochastic mechanisms influence sympatric distributions for these recently diverged species.

Gene flow increases fitness at the warm edge of a species’ range

According to theory, gene flow to marginal populations may stall or aid adaptation at range limits by swamping peripheral populations with maladaptive gene flow or by enhancing genetic variability and reducing inbreeding depression, respectively. We tested these contrasting predictions by manipulating patterns of gene flow of the annual plant, Mimulus laciniatus, at its warm range limit. Gene flow was experimentally applied by using crosses within warm-limit populations (selfed and outcrossed), between warm-limit populations, and between warm-limit and central range populations across two elevational transects. We measured the fitness of offspring in a common garden at the warm-edge species range limit. All sources of gene flow increased seedling emergence at the range limit, suggesting local inbreeding depression at both range limit populations; however, lifetime reproductive success only increased significantly when pollen originated from another warm-limit population. Center-to-warm-edge gene flow was maladaptive by delaying time to development at this warm, fast-drying range limit, whereas edge-to-edge gene flow hastened emergence time and time to reproduction. By empirically testing theory on the effects of gene flow on the formation of geographic range limits, we find benefits of gene flow among populations to be greatest when gene flow is between populations occupying the same range limit. Our results emphasize the overlooked importance of gene flow among populations occurring near the same range limit and highlight the potential for prescriptive gene flow as a conservation option for populations at risk from climate change.

Mating system of four inbreeding monkeyflower (Mimulus) species revealed using ‘progeny‐pair’ analysis of highly informative microsatellite markers

AbstractThe high variability of microsatellite markers has allowed more powerful and novel inferences regarding mating systems. We show that the many alleles at microsatellite loci allow a different approach for estimating a mating system that uses four‐gene coefficients of relationship; the four genes are those possessed by two progeny at a single locus. We estimated the mating system in four Mimulus taxa sampled from California, USA, all of which display inbreeding floral syndromes: Mimulus nasutus, Mimulus micranthus, Mimulus nudatus and Mimulus laciniatus. For each taxa, 20 progeny pairs were assayed for nine microsatellite loci. Substantial amounts of selfing were found, ranging from 64% to 92% across the taxa. Parent inbreeding coefficients were high (range 70%) in two taxa, and in these two taxa both facets of correlated matings (correlation of selfing and of outcrossed paternity) were also much higher (range 50–80%) than in the other two taxa. This association between correlated matings and levels of selfing has not been previously demonstrated. Microsatellites allow new approaches for mating system inference, but at the disadvantage of higher genotyping costs and potential biases resulting from null alleles.

BARRIERS TO GENE EXCHANGE BETWEEN MEMBERS OF THE MIMULUS GUTTATUS COMPLEX (SCROPHULARIACEAE)

A host of stages of active evolution is exhibited by the Mimulus guttatus complex of related species. This group is a living example of Wright's (1932) hypothetical model for rapid evolution and speciation. Its tens of thousands of populations are frequently small and partially or completely geographically isolated from one another just as Wright postulated. The marked morphological and physiological differences between the populations (Grant, 1924; Pennell, 1951) indicate that much evolutionary diversification has al-. ready occurred. However, diversification alone does not lead to the evolution of new species. Speciation requires the development of isolating mechanisms that prevent gene exchange between populations (Dobzhansky, 1951). The evolution of isolating mechanisms may be concurrent with morphological and physiological differentiation, or may be independent of such diversification. The purpose of this study is to identify and investigate those isolating mechanisms which may be present within the Mimulus guttatus complex. The populations selected for investigation well represent the obvious morphological and physiological diversity of the group. They include the main species of the complex (M. guttatus Fischer), its two most common relatives (M. nasutus Greene and M. laciniatus Gray), and two relatively rare members of the group (M. platycalyx Pennell and M. glaucescens Greene). The individual plants as well as the different populations of the species M. guttatus may be from a centimeter to over a meter in height. Their leaves vary in shape and size from one population to the next. The leaves of individual plants range from relatively large and petiolate near the base to comparatively small and sessile at the stem apex (fig. 1). The flowers are conspicuous, yellow usually dotted with red, and strongly bilabiate. The most noticeable difference between M. nasutus and the other members of the complex is its nose-like upper calyx lobe. One M. nasutus population has n = 13 chromosomes instead of n = 14, the characteristic number of the complex (Mukherjee and Vickery, 1960). Mimulus laciniatus differs from the rest of the complex by its deeply laciniate leaves. Mimulus platycalyx is characterized by its open-throated corollas and its cup-shaped to funnel-form calyces. Furthermore, it has n = 15 chromosomes. Mimulus glaucescens can be easily recognized by its connate floral leaves. These are the most conspicuous differences which separate the species chosen for study. The species also differ in other flower and vegetative characteristics as well as in their physiological requirements and adaptations (Grant, 1924; Pennell, 1951). Furthermore, each selected population differs from the others of the same species in smaller, but still obvious morphological and physiological traits (table 1 and fig. 1). For this investigation, the Mimulus 1 Approximately one-fourth of the work reported here was carried out at the Carnegie Institution of Washington's Department of Plant Biology Laboratory, Stanford, California, and was included in the author's dissertation submitted to the faculty of Stanford University in partial fulfillment of the Ph.D. requirements. The rest of the work has been carried out subsequently in the Division of Biology, University of Utah, Salt Lake City, Utah where it has been supported by grants from the University of Utah Research Fund and from the National Science Foundation. The author wishes to thank his colleague, W. W. Newby, for his careful reading of the manuscript and many helpful suggestions.