Mimulus Luteus Research Articles

Endosperm-based incompatibilities in hybrid monkeyflowers.

Endosperm is an angiosperm innovation central to their reproduction whose development, and thus seed viability, is controlled by genomic imprinting, where expression from certain genes is parent-specific. Unsuccessful imprinting has been linked to failed inter-specific and inter-ploidy hybridization. Despite their importance in plant speciation, the underlying mechanisms behind these endosperm-based barriers remain poorly understood. Here, we describe one such barrier between diploid Mimulus guttatus and tetraploid Mimulus luteus. The two parents differ in endosperm DNA methylation, expression dynamics, and imprinted genes. Hybrid seeds suffer from underdeveloped endosperm, reducing viability, or arrested endosperm and seed abortion when M. guttatus or M. luteus is seed parent, respectively, and transgressive methylation and expression patterns emerge. The two inherited M. luteus subgenomes, genetically distinct but epigenetically similar, are expressionally dominant over the M. guttatus genome in hybrid embryos and especially their endosperm, where paternal imprints are perturbed. In aborted seeds, de novo methylation is inhibited, potentially owing to incompatible paternal instructions of imbalanced dosage from M. guttatus imprints. We suggest that diverged epigenetic/regulatory landscapes between parental genomes induce epigenetic repatterning and global shifts in expression, which, in endosperm, may uniquely facilitate incompatible interactions between divergent imprinting schemes, potentially driving rapid barriers.

A likelihood ratio test for changes in homeolog expression bias

BackgroundGene duplications are a major source of raw material for evolution and a likely contributor to the diversity of life on earth. Duplicate genes (i.e., homeologs, in the case of a whole genome duplication) may retain their ancestral function, sub- or neofunctionalize, or be lost entirely. A primary way that duplicate genes evolve new functions is by altering their expression patterns. Comparing the expression patterns of duplicate genes gives clues as to whether any of these evolutionary processes have occurred.ResultsWe develop a likelihood ratio test for the analysis of the expression ratios of duplicate genes across two conditions (e.g., tissues). We demonstrate an application of this test by comparing homeolog expression patterns of 1448 homeologous gene pairs using RNA-seq data generated from leaves and petals of an allotetraploid monkeyflower (Mimulus luteus). We assess the sensitivity of this test to different levels of homeolog expression bias and compare the method to several alternatives.ConclusionsThe likelihood ratio test derived here is a direct, transparent, and easily implemented method for detecting changes in homeolog expression bias that outperforms alternative approaches. While our method was derived with homeolog analysis in mind, this method can be used to analyze changes in the ratio of expression levels between any two genes in any two conditions.

Absence of postmating barriers between a selfing vs. outcrossing Chilean Mimulus species pair.

Reproductive isolation between sympatric species pairs may be maintained by both pre- and postmating barriers. Here we evaluate potential barriers to mating between the outcrossing Mimulus luteus and its more highly selfing sympatric congener, M. cupreus, two members of the South American luteus complex of Mimulus. Seed set was compared following autonomous self-pollination, manual pollination, conspecific outcrossing, and sympatric and allopatric hybridization, for laboratory-maintained inbred lines and wild-collected accessions. Survival and reproductive fitness of hybrids relative to parental species were examined across environments that differed with respect to temperature and soil nutrients, two factors that vary across the ranges of M. luteus and M. cupreus. Mimulus luteus was minimally capable of autonomous self-fertilization, consistent with reliance on an animal pollinator, whereas M. cupreus was a successful selfer across all tested accessions. Postmating barriers to hybridization are negligible, in both low- and high-stress environments, across multiple sympatric and allopatric populations. As in the North American M. guttatus-M. nasutus species pair, postmating barriers contribute little to isolation between M. luteus and M. cupreus. This result reinforces the importance of premating barriers, specifically species differences in reliance on, and accessibility to, animal pollinators. A unique aspect of the M. luteus-M. cupreus pair is the recent gain of red floral anthocyanin pigmentation in M. cupreus. On the basis of species differences in vegetative anthocyanin production, a facultative stress-protective response, we propose a potential stress-protective role for the constitutive floral anthocyanins of M. cupreus.

Pollination ecology and breeding system of two Calceolaria species in Chile

71°00′ W) in the Region del Maule, Chile. The sample site is located in the Reserve at 2,200 m a.s.l. and has a Mediterranean-type climate, with most rainfall concentrated in the winter season (di Castri and Hajek 1976). In the study site, C. filicaulis and C. arachnoidea inhabit in sympatry and are distributed in monospecific patches, surrounded by a vegetation matrix that includes Mimulus luteus Linnaeus (Schrophulareaceae), Mimulus cupreus Dombrain (Schrophulareaceae), Hypochoeris acaulis Britton (Asteraceae), Pozoa coriacea Lagasca & Segura (Apiaceae), and Azorella incisa Weddell (Apiaceae). To determine the breeding system of Calceolaria species, a pollination-controlled experiment was conducted in 120 buds per species. Buds were bagged with a mesh, and each of the following treatments were randomly assigned: (1) control (C), intact flowers exposed to natural pollination; (2) hand cross-pollination (HC), emasculated flowers pollinated with pollen from another plant situated at least 1 m apart; (3) automatic pollination (AP), bagged buds without manipulation; and (4) hand self-pollination (HS), emasculated flowers pollinated by its own pollen. Pollination success was estimated as seed set per plant, and self-incompatibility (ISI) and autogamy (IAS) indexes (Arroyo and Uslar 1993) were estimated as

Assessing the impact of the invasive buff-tailed bumblebee (Bombus terrestris) on the pollination of the native Chilean herb Mimulus luteus

The arrival of exotic pollinators to new habitats may introduce new patterns of floral preference and foraging behavior that modify the structure of the resident plant–pollinator community. The aim of this paper is to examine the potential impact of the exotic bumblebee Bombus terrestris on the pollination service provided by the native pollinator assemblage of the herb Mimulus luteus. The study was performed in a high-elevation locality in the Chilean Andes during the summer seasons of 2010, 2011, and 2012. We recorded visitation rate, and the number of pollen grains transported on the body of B. terrestris and native pollinators and the pollen deposition on the stigmas of M. luteus. Pollinator effectiveness (pollen deposited × visitation rate) was compared among species. Results revealed that B. terrestris was an inefficient pollinator, due to the low amount of pollen delivered on stigmas and the low and intermittent visitation rate across years. The parallel inter-annual variation in the visitation rate of B. terrestris and the native bumblebee Bombus dahlbomii suggests that the integration of B. terrestris had no important consequences for the congeneric species. In general, B. terrestris accounted for a low proportion of the pollen transfer in M. luteus, reaching 4.6 % in 2010, absence of effect in 2011, and 0.01 % in 2012. These results suggest that in spite of being a quickly spreading species in Chile, B. terrestris is still in the initial phase of invasion in this area.

Hybridisation and genetic diversity in introduced Mimulus (Phrymaceae)

Hybridisation among taxa with different ploidy levels is often associated with hybrid sterility. Clonal reproduction can stabilise these hybrids, but pervasive clonality may have a profound impact on the distribution of genetic diversity in natural populations. Here we investigate a widespread triploid taxon resulting from hybridisation between diploid Mimulus guttatus and tetraploid Mimulus luteus, two species that were introduced into the United Kingdom (UK) in the nineteenth century. This hybrid, Mimulus x robertsii, is largely sterile but capable of prolific vegetative propagation and has been recorded in the wild since 1872. We surveyed 40 Mimulus populations from localities across the UK to examine the current incidence of hybrids, and selected seventeen populations for genetic analysis using codominant markers. Cluster analyses revealed two main groups of genetically distinct individuals, corresponding to either diploid (M. guttatus) or polyploid (M. luteus and M. x robertsii) samples. Triploid hybrids were found in around 50% of sampled sites, sometimes coexisting with one of the parental species (M. guttatus). The other parent, M. luteus, was restricted to a single locality. Individual populations of M. x robertsii were genetically variable, containing multiple, highly heterozygous clones, with the majority of genetic variation distributed among- rather than within populations. Our findings demonstrate that this largely sterile, clonal taxon can preserve non-negligible amounts of genetic variation. The presence of genetically variable hybrid populations may provide the material for the continued success of asexual taxa in diverse environments.

Mimulus peregrinus (Phrymaceae): A new British allopolyploid species

Polyploidization plays an important role in species formation as chromosome doubling results in strong reproductive isolation between derivative and parental taxa. In this note I describe a new species, Mimulus peregrinus (Phrymaceae), which represents the first recorded instance of a new British polyploid species of Mimulus (2n = 6x = 92) that has arisen since the introduction of this genus into the United Kingdom in the 1800’s. Mimulus peregrinus presents floral and vegetative characteristics intermediate between Mimulus guttatus and Mimulus luteus, but can be distinguished from all naturalized British Mimulus species and hybrids based on a combination of reproductive and vegetative traits. Mimulus peregrinus displays high pollen and seed fertility as well as traits usually associated with genome doubling such as increased pollen and stomata size. The intermediate characteristics of Mimulus peregrinus between Mimulus guttatus (2n = 2x = 28)and Mimulus luteus (2n = 4x = 60-62), and its close affinity with the highly sterile, triploid (2n = 3x = 44-45) hybrid taxon Mimulus × robertsii (Mimulus guttatus × Mimulus luteus), suggests that Mimulus peregrinus mayconstitute an example of recent allopolyploid speciation.

Cyclohexanones from Mimulus glabratus and M. luteus

The phytochemical study of Mimulus glabratus A.Gray allowed the isolation of two cyclohexenones: the new compound 6-chlorohalleridone 1 and halleridone 2. Halleridone was also identified in Mimulus luteus L., together with dihydroalleridone 3, the naphtoquinone alpha-dunnione 4, ursolic acid and beta-sitosterol.

Effects of herkogamy and inbreeding on the mating system of Mimulus luteus in the absence of pollinators

Self-pollination mechanisms are often invoked to explain plant reproduction in environments devoid of pollinators. However, populations may evolve a diverse variety of strategies depending on the ecological context of pollination. In this study we examined the pollination environment, inbreeding depression, and herkogamy of Mimulus luteus (Phrymaceae) in two consecutive years in an attempt to evaluate the extent to which these factors determine the current level of autogamy in this population. Results indicate that pollinators were almost absent in the study site in 2005 and 2006. Likewise, no evidence of significant inbreeding depression for seed production and seed germination was detected in the 2 years. Because M. luteus presents hermaphrodite flowers with ‘movement herkogamy’ (touch-sensitive stigmata that close upon contact), and anther–stigma separation, we evaluated the role of these traits in self-fertilization. First, we assessed the effects of pollen source (self- and xenogamous pollen, and control solution) on stigmata dynamics after pollen deposition, and seed production. We observed that stigmata that received pollen (self- and xenogamous hand-pollinated) remained closed for a longer time than flowers that received no pollen (control flowers). Seed production, however, was unaffected by pollen source, indicating that movement herkogamy does not prevent self-fertilization in this population. Second, a phenotypic selection analysis revealed that seed production increased with a reduction of anther–stigma separation in absence of pollinators (bagged flowers), suggesting that low herkogamy levels are promoted in this population. Our results indicate that lack of inbreeding depression, ineffective movement herkogamy, and selection against anther–stigma separation are factors that may contribute to the prevalence of autogamous reproductive mechanisms when M. luteus faces environments with scarce mating opportunity.

Is floral diversification associated with pollinator divergence? Flower shape, flower colour and pollinator preference in Chilean Mimulus.

Adaptation to different pollinators is thought to drive divergence in flower colour and morphology, and may lead to interspecific reproductive isolation. Floral diversity was tested for association with divergent pollinator preferences in a group of four closely related wildflower species: the yellow-flowered Mimulus luteus var. luteus and the red-pigmented M. l. variegatus, M. naiandinus and M. cupreus. Patterns of pollinator visitation were evaluated in natural plant populations in central Chile, including both single-species and mixed-species sites. Floral anthocyanin pigments were identified, and floral morphology and nectar variation were quantified in a common garden experiment using seeds collected from the study sites. Mimulus l. luteus, M. l. variegatus and M. naiandinus are morphologically similar and share a single generalist bumblebee pollinator, Bombus dahlbomii. Mimulus cupreus differs significantly from the first three taxa in corolla shape as well as nectar characteristics, and had far fewer pollinator visits. This system shows limited potential for pollinator-mediated restriction of gene flow as a function of flower colour, and no evidence of transition to a novel pollinator. Mimulus cupreus may experience reduced interspecific gene flow due to a lack of bumblebee visitation, but not because of its red pigmentation: rare yellow morphs are equally undervisited by pollinators. Overall, the results suggest that factors other than pollinator shifts may contribute to the maintenance of floral diversity in these Chilean Mimulus species.

The influence of insects and hummingbirds on the geographical variation of the flower phenotype in Mimulus luteus

We describe the geographical variation of corolla and nectar guide size in seven populations of Mimulus luteus (Phrymaceae) in central Chile, and examine whether flower phenotypes associate with taxonomic composition and flower visit patterns of pollinators across populations. Flowers showed higher variation in nectar guide size than corolla size. Mean corolla size increased with the proportion of bees and decreased with the proportion of lepidopterans in the pollinator assemblages. Nectar guide size increased with the proportion of hummingbirds in the pollinator assemblages. When the frequency of flower visits rather than taxonomic composition was considered, the results revealed similar patterns. Because these traits previously have been described as targets of bee‐ and hummingbird‐mediated selection in M. luteus, our results have implications for understanding the processes that determine flower diversification in Chilean Mimulus. Although we cannot rule out ecological sorting as an explanation for the geographical association between pollinators and flower phenotypes, changes in the prevalence and importance of bees and hummingbirds across populations appear to account, at least in part, for the flower phenotypic variation across populations. The extent to which insect and hummingbird pollination in M. luteus produces pollinator‐mediated divergence among populations needs to be examined in future studies.

Nonadditive effects of flower damage and hummingbird pollination on the fecundity of Mimulus luteus

Flower herbivory and pollination have been described as interactive processes that influence each other in their effects on plant reproductive success. Few studies, however, have so far examined their joint effects in natural populations. In this paper we evaluate the influence of flower damage and pollination by the hummingbird Oreotrochilus leucopleurus on the fecundity of the Andean monkey flower Mimulus luteus. We performed a 2x2 factorial experiment, with artificial clipping of lower petals and selective exclusion of the hummingbird as main factors. In spite of the relatively low proportion (27.5%) of the variance in seed production accounted for by the full factorial model, artificial damage and hummingbird exclusion, as well as their interaction, were highly significant, indicating nonadditive effects of factors on plant fecundity. In the presence of hummingbirds, undamaged flowers had a seed production that was 1.7-fold higher than for damaged flowers, suggesting that the effect of flower damage on female reproductive success occurs probably as a consequence of hummingbird discrimination against damaged corollas. This result indicates that the impact of flower herbivory on plant fecundity was contingent on the presence or absence of hummingbirds, suggesting that pollinators may indirectly select for undamaged and probably resistant flower phenotypes. A second interaction effect revealed that undamaged flowers produced 78.5% more seeds in the absence of rather than in the presence of O. leucopleurus, raising the question of the ecological mechanism involved. We suggest that the strong territorial behavior exhibited by the bee Centris nigerrima may confine the foraging activities of the remaining bee species to safe sites within exclosures. Overall, our results provide evidence that hummingbird pollination and flower herbivory have interdependent effects on M. luteus fecundity, which indicates that it will be difficult to predict their ecological and evolutionary consequences unless interactions are analyzed in an integrated form.

Nectar guide fluctuating asymmetry does not relate to female fitness in Mimulus luteus

Most studies assessing the importance of developmental instability of floral characters for pollinator visits and plant fitness have focused on the fluctuating asymmetry (FA) of the corolla phenotype. The importance of stability process for nectar guide characters that signal floral reward has not been considered in the literature. In principle, flowers with symmetrical guides should be more successful at attracting pollinators, therefore increasing their reproductive success in comparison to asymmetrical flowers. In this paper we test this hypothesis in a population of 171 individuals of the Andean monkey flower, Mimulus luteus in northern Chile. This species shows a conspicuous red spot in the landing yellow petal, which permits assessment of the functional relationship between nectar guide FA and female fitness. Our results did not reveal a significant linear nor nonlinear relationship between nectar guide FA and fitness. This result was consistent after controlling the level of FA by guide and corolla size. Because the corolla of M. luteus did not show evidence of UV wavelength reflectance, our negative result could not be attributable to a confounding effect of UV guides. Even though we can not rule out that nectar guide FA correlates better with male than female fitness, the low fraction of the variance in female fitness accounted for nectar guide FA, suggests that other components of the floral phenotype as well as environmental factors may be more important to predict pollinator preference and reproductive success in this species.

VARIABILIDAD EN RASGOS POLINICOS EN EL COMPLEJO MIMULUS LUTEUS L. (PHRYMACEAE) EN CHILE CENTRAL

We quantified the variability of pollen morphology and life history traits among three natural populations of two taxa in the Mimulus luteus complex and a group of putative hybrids that coexist in the Andean ranges of central Chile. Pollen grain production and polar/equatorial length ratio, an index of pollen grain morphology, differed significantly among the taxa. The differences detected in these generally conservative traits, may reflect introgression, known to be common in Mimulus

POLLINATOR-MEDIATED SELECTION ON THE NECTAR GUIDE PHENOTYPE IN THE ANDEAN MONKEY FLOWER, MIMULUS LUTEUS

Mimulus luteus (Scrophulariaceae) is a perennial herb occurring in the South American Andes that shows a wide variation in the size and shape of a red spot on the lower lobe of the yellow flower. We describe the preference of four insects (three bees and one butterfly) and one hummingbird species for floral characters, and estimated the strength, direction, and form of pollinator-mediated selection through female fitness. We applied geometric morphometrics to describe the preference of pollinator species for different guide shapes. Our results revealed striking differences in the floral phenotypes preferred by insects and hummingbirds. Insects visited flowers with corollas 1.25-fold larger and guides 1.72-fold larger than the hummingbird species did. While insects preferred flowers with nectar guides pointing toward the corolla tube, the hummingbird preferred flowers with heart-shaped nectar guides. Most of the floral preferences shown by pollinators translated into significant linear and nonlinear selection coefficients. When selection was analyzed on a per-flower basis and for female fitness, corolla size was under positive directional selection, and nectar guide size and shape were under disruptive selection. Because the insect and hummingbird pollinators showed a strong segregation in their daily activity time, we suggest that current disruptive selection on the nectar guide phenotype can result from the differential availability of the rewarding floral variants over a day. Our findings suggest that pollinator-mediated selection favoring extreme phenotypes in M. luteus may not only contribute to high nectar guide variation found in this species, but also can promote divergence of corolla and nectar guide traits.

Copper tolerance testing in populations of Mimulus luteus var. variegatus exposed and non-exposed to copper mine pollution

Several plant species have evolved metal tolerance as a response to increasing metal concentration in soils due to human activities. A well-described example is the evolution of copper-tolerant ecotypes of the monkey flower ( Mimulus guttatus ) in Copperopolis, California. Although copper mine activities have been intense in some areas of the Andes Range in central Chile, metal-tolerant ecotypes have not been reported for any native or endemic plant species. Copper tolerance of two populations of Mimulus luteus var. variegatus from pristine basins (Río Cipreses and Las Cayanas) and one population from a highly polluted copper mine area (Sewell) were tested to determine their copper tolerance. Tolerance was tested on cuttings collected in the field from adult plants (Sewell population) and on cuttings from plants cultivated from seeds in the laboratory (Río Cipreses, Las Cayanas and Sewell). All cuttings were grown under increasing concentrations of copper (0, 0.5 and 2.0 mg l −1 ) in calcium nitrate (0.5 g l −1 ). Results indicated that root number and length were strongly inhibited in M. luteus var. variegatus from Río Cipreses and Las Cayanas, but tolerant individuals were found in the Sewell population. The degree of copper tolerance of M. luteus var. variegatus from Sewell was, however, lower than that of individuals of Mimulus guttatus from Copperopolis.

Induction of haploid parthenogenesis in Mimulus luteus by in-vitro pollination with foreign pollen

Pistils were isolated from flowers of a clone of Mimulus luteus . The ovary wall was removed and pollen of Torenia fournieri was placed directly onto the ovules. In 1% of the carpels treated in this way a haploid parthenogenesis was induced.

Malonic acid: The acyl moiety of the Mimulus luteus anthocyanin

Malonic acid has been identified as the acylating agent of the anthocyanin produced by the Mimulus luteus complex.

Pattern partitioning of floral flavonoids in the Mimulus luteus complex

Some flavonoids are differentially distributed between the spots and the background in the flowers of the Mimulus luteus complex; others are not. T

Plant polyphenols—XV : Flavonols as yellow flower pigments

The flavonol quercetagetin has been identified as a yellow flower pigment in species of Coronilla, Lotus, Papaver, Primula and Rhododendron; it has been isolated as the 3-galactoside, 3-gentiotrioside or 7-glucoside. It is accompanied in Lotus corniculatus by its 7-methyl ether. A flavonol tentatively identified as the related 6-hydroxykaempferol has been found in Mimulus luteus. Quercetagetin is accompanied in Papaver nudicaule by nudicaulin, a water-soluble yellow pigment earlier considered to be a flavylium salt; spectral studies now indicate that this is unlikely. Syringetin and isorhamnetin have been found as yellow flower pigments in Lathyrus pratensis. While the above compounds, unlike the common flavonols, undoubtedly contribute to yellow flower colour, they accompany yellow carotenoids in many of the plants that have been examined. During the present survey, a water-soluble carotenoid has been found in the petals of only one plant, Nemesia strumosa. This pigment was readily distinguished by spectral means from the yellow flavonols and was identified as crocein.