Evolution of multiple postzygotic barriers between species of the Mimulus tilingii complex.

Species are often defined by their ability to interbreed (i.e., Biological Species Concept), but determining how and why reproductive isolation arises between new species can be challenging. In the Mimulus tilingii species complex, three species (M. caespitosa, M. minor, and M. tilingii) are largely allopatric and grow exclusively at high elevations (>2000 m). The extent to which geographic separation has shaped patterns of divergence among the species is not well understood. In this study, we determined that the three species are morphologically and genetically distinct, yet recently diverged. Additionally, we performed reciprocal crosses within and between the species and identified several strong postzygotic reproductive barriers, including hybrid seed inviability, F1 hybrid necrosis, and F1 hybrid male and female sterility. In this study, such postzygotic barriers are so strong that a cross between any species pair in the M. tilingii complex would cause nearly complete reproductive isolation. We consider how geographical and topographical patterns may have facilitated the evolution of several postzygotic barriers and contributed to speciation of closely related members within the M. tilingii species complex.

Characterization of polymorphic microsatellite loci for invasive wavyleaf basketgrass, Oplismenus undulatifolius (Poaceae).

PREMISE OF THE STUDY: Novel nuclear microsatellite markers were developed for the invasive plant Oplismenus undulatifolius (Poaceae) to facilitate studies of invasion dynamics for this recently introduced, high-risk invasive species in North American mid-Atlantic forests. METHODS AND RESULTS: Sixteen polymorphic microsatellite loci were isolated and characterized from an Illumina paired-end shotgun library of O. undulatifolius after initial assessment of 48 loci. When screened in three populations, these markers identified two to six alleles per locus. Observed heterozygosity ranged from 0.000 to 1.000. All loci were successfully amplified in the North American native congener O. hirtellus. CONCLUSIONS: We provide the first nuclear genetic resources available for population genetic studies within the genus Oplismenus. These markers will serve as a critical tool for exploring patterns of gene flow and mechanisms of invasion of O. undulatifolius across its introduced range. These microsatellites should also be suitable for studies in other Oplismenus species.

Evidence for natural hybridization between native and introduced lineages of Phragmites australis in the Chesapeake Bay watershed.

PREMISE OF THE STUDY: The introduction of nonnative taxa into areas occupied by conspecifics can lead to local extinction of native taxa via habitat modification and competitive dominance, and be exacerbated by outbreeding depression or the formation of invasive hybrid lineages following intraspecific gene flow. The expansion of Eurasian Phragmites australis into tidal wetlands of North America has been accompanied by a dramatic decline of native P. australis, with few relic populations remaining along the Atlantic coastline of the United States, particularly in the Virginia portion of the Chesapeake Bay. METHODS: We sampled populations from the York River and its two major tributaries to determine the pattern of Phragmites invasion and identify remnant native populations that warrant conservation. We used chloroplast DNA haplotypes and nuclear DNA microsatellite profiles to classify individuals as belonging to the native or introduced lineage. KEY RESULTS: Although native Phragmites stands were identified in the brackish upstream reaches of the two York River tributaries, the majority of Phragmites stands surveyed contained the introduced lineage. We also identified a single putative hybrid plant, based on its microsatellite profile. This plant possessed the native cpDNA haplotype and was located in an otherwise native Phragmites stand that is adjacent to an isolated patch of introduced Phragmites. CONCLUSIONS: Although evidence of field hybridization between native and introduced lineages of Phragmites in North America is still relatively rare, the continued encroachment of the introduced lineage into native wetlands may increase the likelihood of future hybrid formation. Careful genetic monitoring to identify remnant native and potential hybrid Phragmites is essential for prioritizing ongoing management efforts.

The genetic basis of a rare flower color polymorphism in Mimulus lewisii provides insight into the repeatability of evolution.

A long-standing question in evolutionary biology asks whether the genetic changes contributing to phenotypic evolution are predictable. Here, we identify a genetic change associated with segregating variation in flower color within a population of Mimulus lewisii. To determine whether these types of changes are predictable, we combined this information with data from other species to investigate whether the spectrum of mutations affecting flower color transitions differs based on the evolutionary time-scale since divergence. We used classic genetic techniques, along with gene expression and population genetic approaches, to identify the putative, loss-of-function mutation that generates rare, white flowers instead of the common, pink color in M. lewisii. We found that a frameshift mutation in an anthocyanin pathway gene is responsible for the white-flowered polymorphism found in this population of M. lewisii. Comparison of our results with data from other species reveals a broader spectrum of flower color mutations segregating within populations relative to those that fix between populations. These results suggest that the genetic basis of fixed differences in flower color may be predictable, but that for segregating variation is not.

Microsatellite loci in Ipomopsis aggregata (Polemoniaceae) and cross-species applicability for ecological genetics studies.

PREMISE OF THE STUDY: Novel microsatellite primers were developed for the native wildflower Ipomopsis aggregata to facilitate ongoing studies of the genetics of local adaptation and patterns of hybridization with closely related species within the genus. METHODS AND RESULTS: Thirteen primer sets were successfully developed and tested using populations from the Rocky Mountains of Colorado, USA. The primers amplified di-, tri-, and tetranucleotide repeats with 1-15 alleles per locus. All primers also amplified fragments with varying success in closely related Ipomopsis species and more distant members of the Polemoniaceae. CONCLUSIONS: The polymorphism levels observed across all loci suggest that these microsatellites may be useful for population genetic studies in Ipomopsis, as well as in studies of other related taxa in the Polemoniaceae.

Natural variation for drought-response traits in the Mimulus guttatus species complex.

Soil moisture is a key factor affecting plant abundance and distribution, both across and within species. In response to water limitation, plants have evolved numerous morphological, physiological, and phenological adaptations. In both well-watered and water-limited conditions, we identified considerable natural variation in drought-related whole-plant and leaf-level traits among closely related members of the Mimulus guttatus species complex that occupy a diversity of habitats in the field. The self-fertilizing Mimulus nasutus and serpentine-endemic Mimulus nudatus demonstrated the overall greatest tolerance to soil water limitation, exhibiting the smallest reduction in seed set relative to well-watered conditions. This may be due in part to early flowering, faster fruit development, and low stomatal density. In contrast, flowering of coastal M. guttatus was so delayed that it precluded any seed production in water-limited conditions. This range of phenotypic responses to soil water deficit in Mimulus, coupled with developing genomic resources, holds considerable promise for identifying genomic variation responsible for adaptive responses to soil water availability.

Photosynthetic and growth responses of reciprocal hybrids to variation in water and nitrogen availability.

PREMISE OF THE STUDY: Fitness of plant hybrids often depends upon the environment, but physiological mechanisms underlying the differential responses to habitat are poorly understood. We examined physiological responses of Ipomopsis species and hybrids, including reciprocal F(1)s and F(2)s, to variation in soil moisture and nitrogen. • METHODS: To examine responses to moisture, we subjected plants to a dry-down experiment. Nitrogen was manipulated in three independent experiments, one in the field and two in common environments. • KEY RESULTS: Plants with I. tenuituba cytoplasmic background had lower optimal soil moisture for photosynthesis, appearing better adapted to dry conditions, than plants with I. aggregata cytoplasm. This result supported a prediction from prior studies. The species and hybrids did not differ greatly in physiological responses to nitrogen. An increase in soil nitrogen increased leaf nitrogen, carbon assimilation, integrated water-use efficiency, and growth, but the increases in growth were not mediated primarily by an increase in photosynthesis. In neither the field, nor in common-garden studies, did physiological responses to soil nitrogen differ detectably across plant types, although only I. aggregata and hybrids increased seed production in the field. • CONCLUSIONS: These results demonstrate differences in photosynthetic responses between reciprocal hybrids and suggest that water use is more important than nitrogen in explaining the relative photosynthetic performance of these hybrids compared to their parents.

Lifetime fitness in two generations of Ipomopsis hybrids.

Various models purporting to explain natural hybrid zones make different assumptions about the fitness of hybrids. One class of models assumes that hybrids have intrinsically low fitness due to genetic incompatibilities, whereas other models allow hybrid fitness to vary across natural environments. We used the intrinsic rate of increase to assess lifetime fitness of hybrids between two species of montane plants Ipomopsis aggregata and Ipomopsis tenuituba planted as seed into multiple field environments. Because fitness is predicted to depend upon genetic composition of the hybrids, we included F1 hybrids, F2 hybrids, and backcrosses in our field tests. The F2 hybrids had female fitness as high, or higher, than expected under an additive model of fitness. These results run counter to any model of hybrid zone dynamics that relies solely on intrinsic nuclear genetic incompatibilities. Instead, we found that selection was environmentally dependent. In this hybrid zone, cytoplasmic effects and genotype-by-environment interactions appear more important in lowering hybrid fitness than do intrinsic genomic incompatibilities between nuclear genes.

Review. The strength and genetic basis of reproductive isolating barriers in flowering plants.

Speciation is characterized by the evolution of reproductive isolation between two groups of organisms. Understanding the process of speciation requires the quantification of barriers to reproductive isolation, dissection of the genetic mechanisms that contribute to those barriers and determination of the forces driving the evolution of those barriers. Through a comprehensive analysis involving 19 pairs of plant taxa, we assessed the strength and patterns of asymmetry of multiple prezygotic and postzygotic reproductive isolating barriers. We then reviewed contemporary knowledge of the genetic architecture of reproductive isolation and the relative role of chromosomal and genic factors in intrinsic postzygotic isolation. On average, we found that prezygotic isolation is approximately twice as strong as postzygotic isolation, and that postmating barriers are approximately three times more asymmetrical in their action than premating barriers. Barriers involve a variable number of loci, and chromosomal rearrangements may have a limited direct role in reproductive isolation in plants. Future research should aim to understand the relationship between particular genetic loci and the magnitude of their effect on reproductive isolation in nature, the geographical scale at which plant speciation occurs, and the role of different evolutionary forces in the speciation process.

Leaf physiology reflects environmental differences and cytoplasmic background in Ipomopsis (Polemoniaceae) hybrids.

Natural hybridization can produce individuals that vary widely in fitness, depending upon the performance of particular genotypes in a given environment. In a hybrid zone with habitat heterogeneity, differences in physiological responses to abiotic conditions could influence the fitness and spatial distribution of hybrids and parental species. This study compared gas exchange physiology of Ipomopsis aggregata, I. tenuituba, and their natural hybrids in situ and assessed whether physiological differences were consistent with their native environmental conditions. We also produced reciprocal F2s in a greenhouse study to test for cytonuclear effects on water-use efficiency (WUE). The relative performance of natural hybrids and parentals was consistent with their native habitats: I. aggregata at the coolest, wettest locations had the lowest WUE, while hybrids from the most xeric sites had the highest WUE. In hybrids, the mechanism by which both natural and experimental hybrids achieved this high WUE depended on cytotype: those with I. tenuituba cytoplasm had reduced transpiration, while those with I. aggregata cytoplasm had an increased photosynthetic rate, consistent with patterns in the cytoplasmic parent. The high WUE in hybrids may contribute to their high survival in the dry center of the natural hybrid zone, consistent with environment-dependent models of hybrid zone dynamics.

Environmental stressors differentially affect leaf ecophysiological responses in two Ipomopsis species and their hybrids.

The recombination that follows natural hybridization may produce hybrid genotypes with traits that are intermediate or extreme relative to the parental species, and these traits may influence the relative fitness of the hybrids. Here we examined leaf ecophysiological traits that may influence fitness patterns in a natural plant hybrid zone. We compared the biochemical photosynthetic capacity of Ipomopsis aggregata, I. tenuituba, and early generation hybrids, as well as their photosynthetic responses to varying light and temperature, two abiotic factors found to differ among sites along the hybrid zone. In general, ecophysiological traits expressed in these plants were consistent with their natural habitat, even when grown under common greenhouse conditions. I. tenuituba reached higher photosynthetic rates (A) at higher light levels than I. aggregata, and also had a higher optimal temperature for photosynthesis (Topt). This suite of traits may reflect adaptations to the more exposed, rocky sites where I. tenuituba is found, compared to the more vegetated, mesic I. aggregata site. Hybrids had characters that were largely intermediate or tenuituba-like, but particular individual hybrids were extreme for some traits, including light saturation level, light-saturated A, and Topt. Many of these traits are consistent with adaptations reported for plants found in warm, dry sites, so they may put certain hybrids at an advantage at the relatively xeric center of the natural hybrid zone.

Ecophysiology of first and second generation hybrids in a natural plant hybrid zone.

Hybrids between related species vary widely in relative fitness, and that fitness can depend upon the environment. We investigated aspects of physiology that might influence fitness patterns in a plant hybrid zone. Seeds of Ipomopsis aggregata, I. tenuituba, F1 hybrids, F2 hybrids, and offspring of crosses between natural hybrids were planted into the relatively mesic site of origin for I. aggregata and the drier site for natural hybrids. We measured rates of photosynthesis (Amax), transpiration (E), instantaneous (A/E) and long-term (delta13C) indices of water use efficiency (WUE), and leaf nitrogen and carbon. We also examined correlations of these traits with plant size. Photosynthetic rate and A/E were higher in vegetative than flowering plants. WUE varied between sites and years, but differences among genotypic classes were spatially and temporally consistent. Instantaneous WUE was higher for F1 hybrids than for the average of the parental species, thereby showing heterosis. There was no evidence of hybrid breakdown, as WUE was no different in the F2 than the average across the F1 and parental species. Nor did WUE depend on cross direction in producing F1 progeny. Carbon isotope discrimination revealed higher long-term water use efficiency in I. tenuituba than I. aggregata. Leaf nitrogen was higher in I. tenuituba than I. aggregata, and higher in offspring of natural hybrids than in the F2. Results indicate heterosis for water use efficiency, with no hybrid breakdown. Heterosis in WUE may help to explain the relatively high survival of both reciprocal F1 hybrids in dry sites within the natural hybrid zone.

Cytoplasmic and nuclear markers reveal contrasting patterns of spatial genetic structure in a natural Ipomopsis hybrid zone.

Spatial variation in natural selection may play an important role in determining the genetic structure of hybridizing populations. Previous studies have found that F1 hybrids between naturally hybridizing Ipomopsis aggregata and Ipomopsis tenuituba in central Colorado differ in fitness depending on both genotype and environment: hybrids had higher survival when I. aggregata was the maternal parent, except in the centre of the hybrid zone where both hybrid types had high survival. Here, we developed both maternally (cpDNA PCR-RFLP) and biparentally inherited (nuclear AFLP) species-diagnostic markers to characterize the spatial genetic structure of the natural Ipomopsis hybrid zone, and tested the prediction that the majority of natural hybrids have I. aggregata cytoplasm, except in areas near the centre of the hybrid zone. Analyses of 352 individuals from across the hybrid zone indicate that cytoplasmic gene flow is bidirectional, but contrary to expectation, most plants in the hybrid zone have I. tenuituba cytoplasm. This cytotype distribution is consistent with a hybrid zone in historical transition, with I. aggregata nuclear genes advancing into the contact zone. Further, nuclear data show a much more gradual cline than cpDNA markers that is consistent with morphological patterns across the hybrid populations. A mixture of environment- and pollinator-mediated selection may contribute to the current genetic structure of this hybrid system.