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Patterns of nucleotide diversity in two species of Mimulus are affected by mating system and asymmetric introgression.

The evolutionary transition from outcrossing to self-fertilization has far-reaching implications for patterns of intraspecific genetic diversity and the potential for speciation. Using DNA sequence variation at two nuclear loci, we examined the divergence history of two closely related species of Mimulus. To investigate the effects of mating system and introgressive hybridization on the outcrossing M. guttatus and the selfing M. nasutus, we inspected nucleotide diversity within and between natural populations spanning the species' geographic ranges. High sequence similarity among populations of the selfing M. nasutus points to a single evolutionary origin for the species. Consistent with their distinct mating systems, all genetic variation in M. nasutus is distributed among populations, whereas M. guttatus exhibits appreciable levels of nucleotide diversity within populations. Silent genetic diversity is extensive in M. guttatus (mean theta(sil)/site = 0.077) and greatly exceeds the predicted twofold elevation in neutral variation for outcrossers relative to selfers. The finding of several M. guttatus sequences that share complete identity with sequences from M. nasutus suggests that recent asymmetric introgression may have occurred. We argue that exceptionally high nucleotide diversity in M. guttatus is consistent with a long-term history of directional introgression from M. nasutus to M. guttatus throughout the divergence of these two species.

Base Sequence↗

Environment and pollinator-mediated selection on parapatric floral races of Mimulus aurantiacus.

We tested whether selection by pollinators could explain the parapatric distribution of coastal red- and inland yellow-flowered races of Mimulus aurantiacus (Phrymaceae) by examining visitation to natural and experimental populations. As a first step in evaluating whether indirect selection might explain floral divergence, we also tested for local adaptation in early life stages using a reciprocal transplant experiment. Hummingbirds visited flowers of each race at similar rates in natural populations but showed strong (>95%) preference for red flowers in all habitats in experimental arrays. Hawkmoths demonstrated nearly exclusive (>99% of visits) preference for yellow flowers and only visited in inland regions. Strong preferences for alternative floral forms support a direct role for pollinators in floral divergence. Despite these preferences, measures of plant performance across environments showed that red-flowered plants consistently survived better, grew larger and received more overall pollinator visits than yellow-flowered plants. Unmeasured components of fitness may favour the yellow race in inland habitats. Alternatively, we document a marked recent increase in inland hummingbird density that may have caused a change in the selective environment, favouring the eastward advance of red-flowered plants.

Animals↗

The potential for adaptive evolution of pollen grain size in Mimulus guttatus.

We tested whether pollen grain size (PGS) shows heritable variation in three independent populations of Mimulus guttatus by imposing artificial selection for this character. In addition, we looked for correlated responses to selection in a range of 15 other floral characters. Heritable variation in PGS was found in all three populations, with heritabilities of between 19 and 40% (average 30%). After three generations, upward and downward lines differed on average by 30% in pollen volume. No consistent patterns of correlated response were found in other characters, indicating that PGS can respond to selective forces acting on PGS alone. Possible selection mechanisms on PGS in this species could include intermale selection, if large pollen grains produce more competitive gametophytes; or optimization of patterns of resource allocation, if local mate competition varies.

Adaptation, Physiological↗

A novel meiotic drive locus almost completely distorts segregation in mimulus (monkeyflower) hybrids.

We report the discovery, mapping, and characterization of a meiotic drive locus (D) exhibiting nearly 100% nonrandom transmission in hybrids between two species of yellow monkeyflowers, outcrossing Mimulus guttatus and selfing M. nasutus. Only 1% of F(2) hybrids were M. nasutus homozygotes at the marker most tightly linked to D. We used a set of reciprocal backcrosses to distinguish among male-specific, female-specific, and zygote-specific sources of transmission ratio distortion. Transmission was severely distorted only when the heterozygous F(1) acted as the female parent in crosses to either parental species, ruling out pollen competition and zygote mortality as potential sources of drive. After four generations of backcrossing to M. nasutus, nearly isogenic lines were still >90% heterozygous at markers linked to D, suggesting that heterozygosity at the drive locus alone is sufficient for nonrandom transmission. A lack of dramatic female fitness costs in these lines rules out alternatives involving ovule or seed mortality and points to a truly meiotic mechanism of drive. The strength and direction of drive in this system is consistent with population genetic theory of selfish element evolution under different mating systems. These results are the first empirical demonstration of the strong female-specific drive predicted by new models of selfish centromere turnover.

Chimera↗

Demography of central and marginal populations of monkeyflowers (Mimulus cardinalis and M. lewisii).

Every species occupies a limited geographic area, but how spatiotemporal environmental variation affects individual and population fitness to create range limits is not well understood. Because range boundaries arise where, on average, populations are more likely to go extinct than to persist, range limits are an inherently population-level problem for which a demographic framework is useful. In this study, I compare demographic parameters and population dynamics between central and marginal populations of monkeyflowers, Mimulus cardinalis and M. lewisii, along an elevation gradient spanning both species' ranges. Central and marginal populations of both species differed in survival and fecundity. For M. lewisii, these components of fitness were higher in central than in marginal populations, but for M. cardinalis the converse was true. To assess spatiotemporal variation in population dynamics, I used transition matrix models to estimate asymptotic population growth rates (lambda) and found that population growth rates of M. lewisii were highest at the range center and reduced at the range margin. Population growth rates of M. cardinalis were highest at the range margin and greatly reduced at the range center. Life table response analysis decomposed spatiotemporal variation in lambda into contributions from each transition between life stages, finding that transitions from large nonreproductive and reproductive plants to the seed class and stasis in the reproductive class made the largest contributions to spatial differences in lambda. These transitions had only low to moderate sensitivities, indicating that differences in projected population growth rates resulted mainly from observed differences in transition matrix parameters and their underlying vital rates.

Altitude↗

Individual and combined effects of Ca/Mg ratio and water on trait expression in Mimulus guttatus.

Low Ca/Mg ratios (a defining component of serpentine soils) and low water environmental conditions often co-occur in nature and are thought to exert strong selection pressures on natural populations. However, few studies test the individual and combined effects of these environmental factors. We investigated the effects of low Ca/Mg ratio and low water availability on plant leaf, stem, stolon, and floral traits of Mimulus guttatus, a bodenvag species, i.e., a species that occurs in serpentine and non-serpentine areas. We quantified genetic variation and genetic variation for plasticity for these leaf, stem, stolon, and floral traits at three hierarchical levels: field-habitat type, population, and family, and we evaluated the relative importance of local adaptation and plasticity. We chose two populations and 10 families per population from four distinct field "habitat types" in northern California: high Ca/Mg ratio (non-serpentine) and season-long water availability, high Ca/Mg ratio and seasonally drying, low Ca/Mg ratio (serpentine) and season-long water availability, and low Ca/Mg ratio and seasonally drying. Seedlings were planted into greenhouse treatments that mimicked the four field conditions. We only detected genetic variation for stem diameter and length of longest leaf at the field-habitat level, but we detected genetic variation at the family level for nearly all traits. Soil chemistry and water availability had strong phenotypic effects, alone and in combination. Our hypothesis of an association between responses to low water levels and low Ca/Mg ratio was upheld for length of longest leaf, stem diameter, corolla width, and total number of reproductive units, whereas for other traits, responses to Ca/Mg ratio and low water were clearly independent. Our results suggest that traits may evolve independently from Ca/Mg ratios and water availability and that our focal traits were not simple alternative measures of vigor. We found genetic variation for plasticity both at the field-habitat type and family levels for half of the traits studied. Phenotypic plasticity and genetic variation for plasticity appear to be more important than local adaptation in the success of these M. guttatus populations found across a heterogeneous landscape in northern California. Phenotypic plasticity is an important mechanism maintaining the broad ecological breadth of native populations of M. guttatus.

Adaptation, Physiological↗

The evolution of species' distributions: reciprocal transplants across the elevation ranges of Mimulus cardinalis and M. lewisii.

Every species occupies a limited geographic area, but it remains unclear why traits that limit distribution do not evolve to allow range expansion. Hypotheses for the evolutionary stability of geographic ranges assume that species are maladapted at the range boundary and unfit beyond the current range, but this assumption has rarely been tested. To examine how fitness varies across species' ranges, we reciprocally transplanted two species of monkeyflowers, Mimulus cardinalis and M. lewisii, within and beyond their present elevation ranges. We used individuals of known parentage from populations collected across the elevation ranges of both species to examine whether populations are adapted to position within the range. For both species we found the greatest average fitness at elevations central within the range, reduced fitness at the range margin, and zero or near-zero fitness when transplanted beyond their present elevation range limits. However, the underlying causes of fitness variation differed between the species. At high elevations beyond its range, M. cardinalis displayed reduced growth and fecundity, whereas at low elevations M. lewisii experienced high mortality. Weak differences in performance were observed among populations within each species and these were not related to elevation of origin. Low fitness of both species at their range margin and weak differentiation among populations within each species suggest that adaptation to the environment at and beyond the range margin is hindered, illustrating that range margins provide an interesting system in which to study limits to adaptation.

Adaptation, Biological↗

Contrasting patterns of floral and molecular variation across a cline in Mimulus aurantiacus.

Steep clines in ecologically important traits may be caused by divergent natural selection. However, processes that do not necessarily invoke ongoing selection, such as secondary contact or restricted gene flow, can also cause patterns of phenotypic differentiation over short spatial scales. Distinguishing among all possible scenarios is difficult, but an attainable goal is to establish whether scenarios that imply selection need to be invoked. We compared the extent of morphological and genetic differentiation between geographically structured red and yellow floral races of Mimulus aurantiacus (bush monkeyflower; Phrymaceae). Flower color was assessed in a common garden as well as in the field to determine whether variation was genetic and to quantify the extent of geographical differentiation. Population genetic differentiation at marker loci was measured for both chloroplast and nuclear genomes, and the degree of population structure within and among the floral races was evaluated. Flower color shows both a strong genetic basis and a sharp geographic transition, with pure red-flowered populations in western San Diego County and pure yellow-flowered populations to the east. In the zone of contact, both pure and intermediate phenotypes occur. Patterns of genetic differentiation at marker loci are far less pronounced, as little of the variation is partitioned according to the differences in flower color. Phenotypic differentiation (Q(ST)) between populations with different flower colors is much greater than neutral genetic differentiation (F(ST)). When comparisons are made between populations of the same flower color, the opposite trend is evident. Limited neutral genetic structure between the floral races, combined with sharp differentiation in flower color, is consistent with the hypothesis that current or recent natural selection maintains the cline in flower color.

Biological Evolution↗

Metallothionein genes from the flowering plant Mimulus guttatus.

In response to excess metal, higher plants produce metal-binding peptides [( gamma EC]nG) whose biosynthesis is believed to be mediated by enzymes involved in glutathione (gamma ECG) metabolism. In contrast, animals synthesize metallothioneins, gene-encoded low molecular weight cysteine-rich metal-binding proteins. In an investigation of copper-regulated genes in the copper-tolerant flowering plant Mimulus guttatus, we have isolated a series of cDNA clones identifying two genes which encode a protein with class I metallothionein domains. This represents the first description of a metallothionein gene in a flowering plant.

Cloning, Molecular↗

Mimulus finds centromeres in the driver's seat.

Sandler and Novitski first pointed out in 1957 that chromosomes could selfishly exploit meiotic asymmetries to maximize their own transmission, in a process termed 'meiotic drive'. However, since then, only post-meiotic processes of non-Mendelian inheritance have received serious scientific attention in studies of transmission distortion. A recent study by Fishman and Willis puts the focus squarely back on meiotic drive. They found completely biased transmission of a centromere-linked locus from an outcrossing Mimulus species over that from an inbred species, providing the first direct evidence that centromeres can act as general, powerful meiotic drivers. This study suggests that, although difficult to detect experimentally, female meiotic drive is a major evolutionary force in nature.

Journal Article↗

Structure of a geranyl-alpha-pyrone from Mimulus aurantiacus leaf resin.

A previously reported but misidentified geranyl-alpha-pyrone, in addition to six known compounds, was isolated from the leaf resin of Mimulus aurantiacus. HMBC NMR analyses of the geranyl-alpha-pyrone resolved uncertainties in the site of attachment of the two side chains and necessitated a revision of the previously reported structure. This compound is shown to be 3-geranyl-4-hydroxy-6-(2-hydroxypropyl)-2-pyrone.

Magnetic Resonance Spectroscopy↗

C-geranyl compounds from Mimulus clevelandii.

Fractionation of the MeCOEt extract of Mimulus clevelandii yielded the novel 4-geranyl-5-hydroxy-2(3H)-benzofuranone (1) and the five known 6-geranylflavanones diplacone (2a), 3'-O-methyldiplacone (2b), diplacol (2c), mimulone (2d), and 3'-O-methyldiplacol (2e). 2D-NMR methods required revision of assignments for diplacone and diplacol and resolved the uncertainty in the site of methylation for the methyl ethers.

Acyclic Monoterpenes↗

The contribution of male-sterility mutations to inbreeding depression in Mimulus guttatus.

The magnitude of inbreeding depression can influence many aspects of a population's ecology and evolution, including the nature of selection acting on the mating system and the chances that the population will go extinct during periods of small population size. If inbreeding depression is caused primarily by recessive mutations of large effect on fitness, such as lethals or steriles, then it is expected to be purged rapidly by selection with moderate amounts of inbreeding. In contrast, inbreeding depression primarily caused by many genes with mild effects on fitness will not be rapidly purged with inbreeding, so it should be a more resilient barrier to the evolution of self-fertilization and a more significant threat to the survival of endangered species. Here I show that recessive male-sterility alleles at individual loci are common in a primarily outcrossing population of the plant Mimulus guttatus. Despite the high frequency of these major mutations, most of the inbreeding depression for male fertility and cumulative measures of lifetime fitness results from more mildly deleterious alleles. Male-sterility alleles contribute to 31% of the inbreeding depression for the fraction of viable pollen grains, and to 26% of the inbreeding depression for total fitness. These results suggest that most of the inbreeding depression for male fertility in this population would not be purged, in the short term, with moderate inbreeding.

Alleles↗

The distribution of individual inbreeding coefficients and pairwise relatedness in a population of mimulus guttatus

In order to infer population structure at the individual level, we estimated individual inbreeding coefficients and examined the relationship between geographical distance and genetic relatedness from polymorphic microsatellite data for a population of Mimulus guttatus that has an intermediate selfing rate. Expected heterozygosities for five microsatellites ranged from 0.79 to 0.93. The population inbreeding coefficient was calculated to be 0.19 (SE=0.023). A method-of-moments estimator developed by Ritland (1996b) was used to estimate the distribution of inbreeding among and relatedness between individuals of a natural population. The mean individual inbreeding coefficient (F=0.16) did not differ significantly from the population-level estimate. Most of the individuals appeared to be outbred, and there were very few plants that had estimated inbreeding coefficients greater than one-half. Individuals sampled from one transect showed significantly more inbreeding than individuals sampled along the other (P=0.005). There was no apparent relationship between interplant distance (range: 0-14 m) and mean genetic relatedness between individuals. These results represent the first application of polymorphic microsatellites to estimate fine-scale genetic population structure.

Journal Article↗

Evidence for the partial dominance of viability genes contributing to inbreeding depression in Mimulus guttatus.

The relative importance of different modes of gene expression of viability genes contributing to inbreeding depression was investigated in the wild plant, Mimulus guttatus. Viability genes were identified by self-fertilizing 31 outbred plants, each heterozygous for three to nine unlinked allozyme markers, and analyzing segregation ratios of selfed progeny at maturity for deviations from 1:2:1 ratios. In this study, 24 linkages of viability genes to marker loci were detected. To infer the nature of gene action for these viability genes, a "model-free" graphical method was developed that examines the "space" of segregation ratios allowed by each of seven selection models (i.e., overdominance, complete recessivity, partial recessivity, additivity, partial dominance, complete dominance and underdominance). Using this method, we found that, of 24 linkages detected, 18 were consistent with either partial dominance, complete dominance or underdominance. Six were consistent with either partial recessivity, complete recessivity or overdominance. This finding indicates that, in these chromosomal segments identified by allozyme markers, partial dominance plays the predominant role in inbreeding depression. This is inconsistent with either the dominance or overdominance hypotheses proposed to account for inbreeding depression.

Crosses, Genetic↗

Quantitative trait loci differentiating the outbreeding Mimulus guttatus from the inbreeding M. platycalyx.

Theoretical predictions about the evolution of selfing depend on the genetic architecture of loci controlling selfing (monogenic vs. polygenic determination, large vs. small effect of alleles, dominance vs. recessiveness), and studies of such architecture are lacking. We inferred the genetic basis of mating system differences between the outbreeding Mimulus guttatus and the inbreeding M. platycalyx by quantitative trait locus (QTL) mapping using random amplified polymorphic DNA and isozyme markers. One to three QTL were detected for each of five mating system characters, and each QTL explained 7.6-28.6% of the phenotypic variance. Taken together, QTL accounted for up to 38% of the variation in mating system characters, and a large proportion of variation was unaccounted for. Inferred QTL often affected more than one trait, contributing to the genetic correlation between those traits. These results are consistent with the hypothesis that quantitative variation in plant mating system characters is primarily controlled by loci with small effect.

Chromosome Mapping↗

Quantitative trait loci affecting differences in floral morphology between two species of monkeyflower (Mimulus).

Conspicuous differences in floral morphology are partly responsible for reproductive isolation between two sympatric species of monkeyflower because of their effect on visitation of the flowers by different pollinators. Mimulus lewisii flowers are visited primarily by bumblebees, whereas M. cardinalis flowers are visited mostly by hummingbirds. The genetic control of 12 morphological differences between the flowers of M. lewisii and M. cardinalis was explored in a large linkage mapping population of F2 plants n = 465 to provide an accurate estimate of the number and magnitude of effect of quantitative trait loci (QTLs) governing each character. Between one and six QTLs were identified for each trait. Most (9/12) traits appear to be controlled in part by at least one major QTL explaining >/=25% of the total phenotypic variance. This implies that either single genes of individually large effect or linked clusters of genes with a large cumulative effect can play a role in the evolution of reproductive isolation and speciation.

Chromosome Mapping↗

Inbreeding load, average dominance and the mutation rate for mildly deleterious alleles in Mimulus guttatus.

The goal of this study is to provide information on the genetics of inbreeding depression in a primarily outcrossing population of Mimulus guttatus. Previous studies of this population indicate that there is tremendous inbreeding depression for nearly every fitness component and that almost all of this inbreeding depression is due to mildly deleterious alleles rather than recessive lethals or steriles. In this article I assayed the homozygous and heterozygous fitnesses of 184 highly inbred lines extracted from a natural population. Natural selection during the five generations of selfing involved in line formation essentially eliminated major deleterious alleles but was ineffective in purging alleles with minor fitness effects and did not appreciably diminish overall levels of inbreeding depression. Estimates of the average degree of dominance of these mildly deleterious alleles, obtained from the regression of heterozygous fitness on the sum of parental homozygous fitness, indicate that the detrimental alleles are partially recessive for most fitness traits, with h approximately 0.15 for cumulative measures of fitness. The inbreeding load, B, for total fitness is approximately 1.0 in this experiment. These results are consistent with the hypothesis that spontaneous mildly deleterious mutations occur at a rate >0.1 mutation per genome per generation.

Alleles↗