Search

Floral morphology and pigmentation are both charismatic and economically relevant traits associated with cultivated sunflower (Helianthus annuus L.). Recent work has linked floral morphology and pigmentation to pollinator efficiency and seed yield. Understanding the genetic architecture of such traits is essential for crop improvement, and gives insight into the role of genetic constraints in shaping floral diversity. A diversity panel of 288 sunflower genotypes was phenotyped for a variety of morphological, phenological, and color traits in both a greenhouse and a field setting. Association mapping was performed using 5788 SNP markers using a mixed linear model approach. Several dozen markers across 10 linkage groups were significantly associated with variation in morphological and color trait variation. Substantial trait plasticity was observed between greenhouse and field phenotyping, and associations differed between environments. Color traits mapped more strongly than morphology in both settings, with markers together explaining 16% of petal carotenoid content in the greenhouse, and 17% and 24% of variation in disc anthocyanin presence in the field and greenhouse, respectively. Morphological traits like disc size mapped more strongly in the field, with markers together explaining up to 19% of disc size variation. Loci identified here through association mapping within cultivated germplasm differ from those identified through biparental crosses between modern cultivated sunflower and either its wild progenitor or domesticated landraces. Several loci lie within genomic regions involved in domestication. Differences between phenotype expression under greenhouse and field conditions highlight the importance of plasticity in determining floral morphology and pigmentation.

With rising food demands, crop production on salinized lands is increasingly necessary. Sunflower (Helianthus annuus), a moderately salt-tolerant crop, exhibits a tradeoff where more vigorous, high-performing genotypes have a greater proportional decline in biomass under salinity stress. Prior research has found deviations from this relationship across genotypes. Here, we identified the traits and genomic regions underlying variation in this expectation-deviation tolerance (the magnitude and direction of deviations from the expected effect of salinity). We grew a sunflower diversity panel under control and salt-stressed conditions and measured a suite of morphological (growth, mass allocation, plant and leaf morphology) and leaf ionomic traits. The genetic basis of variation and plasticity in these traits was investigated via genome-wide association, which also enabled the identification of genomic regions (i.e. haplotypic blocks) influencing multiple traits. We found that the magnitude and direction of plasticity in whole-root mass fraction, fine root mass fraction, and chlorophyll content, as well as leaf sodium and potassium content under saline conditions, were most strongly correlated with expectation-deviation tolerance. We identified multiple genomic regions underlying these traits as well as a single alpha-mannosidase gene directly associated with this tolerance metric. Our results show that, by taking the vigor-salinity effect tradeoff into account, we can identify unique traits and genes associated with salinity tolerance. Since these traits and genomic regions are distinct from those associated with high vigor (i.e. growth in benign conditions), they provide an avenue for increasing salinity tolerance in high-performing sunflower genotypes without compromising vigor.

Species often include multiple ecotypes that are adapted to different environments 1. However, it is unclear how ecotypes arise and how their distinctive combinations of adaptive alleles are maintained despite hybridization with non-adapted populations 2-4. Here, by resequencing 1,506 wild sunflowers from 3 species (Helianthus annuus, Helianthus petiolaris and Helianthus argophyllus), we identify 37 large (1-100 Mbp in size), non-recombining haplotype blocks that are associated with numerous ecologically relevant traits, as well as soil and climate characteristics. Limited recombination in these haplotype blocks keeps adaptive alleles together, and these regions differentiate sunflower ecotypes. For example, haplotype blocks control a 77-day difference in flowering between ecotypes of the silverleaf sunflower H. argophyllus (probably through deletion of a homologue of FLOWERING LOCUS T (FT)), and are associated with seed size, flowering time and soil fertility in dune-adapted sunflowers. These haplotypes are highly divergent, frequently associated with structural variants and often appear to represent introgressions from other-possibly now-extinct-congeners. These results highlight a pervasive role of structural variation in ecotypic adaptation. Local adaptation is common in species that experience different environments across their range, often resulting in the formation of ecotypes-ecological races with distinct morphological and/or physiological characteristics that provide an environment-specific fitness advantage. Despite the prevalence of ecotypic differentiation, much remains to be understood about the genetic basis and evolutionary mechanisms that underlie its establishment and maintenance. In particular , a longstanding evolutionary question-dating to criticisms of Darwin's theories by his contemporaries 4-concerns how such ecological divergence can occur when challenged by hybridization with non-adapted populations 2. Local adaptation typically requires alleles at multiple loci that contribute to increased fitness in the same environment ; however, different ecotypes are often geographically close and interfertile, and hybridization between them should break up adaptive allelic combinations 3. To better understand the genetic basis of local adaptation and ecotypic differentiation, we conducted an in-depth study of genetic, phenotypic and environmental variation in three annual sunflower species, each of which includes multiple reproductively compatible ecotypes. Two species (H. annuus and H. petiolaris) have broad, overlapping distributions across North America. Helianthus annuus, the common sunflower, is generally found on mesic soils, but can grow in a variety of disturbed or extreme habitats, including semi-desert or frequently flooded areas. An especially well-characterized ecotype (formally known as H. annuus subsp. texanus) is adapted to the higher temperatures and herbivore pressures in Texas (USA) 5. Helianthus petiolaris, the prairie sunflower, prefers sandier soils; ecotypes of this species are adapted to sand sheets and dunes 6. The third species-H. argophyllus, the silverleaf sunflower-is endemic to southern Texas and includes both an early-flowering, coastal-island ecotype and a late-flowering inland ecotype 7. Population structure of wild sunflowers In a common garden experiment, we grew 10 plants from each of 151 populations of the 3 species, selected from across their native range (Fig. 1a); for each of these populations, we collected corresponding soil samples. We generated extensive records of developmental and morphological traits, and resequenced the genomes of 1,401 individual plants. We resequenced an additional 105 H. annuus plants to fill gaps in geographical coverage, as well as 12 outgroup taxa (Supplementary Table 1). Sunflower genomes are relatively large (H. annuus, 3.5 Gbp; https://doi.

Foliar nutrient resorption is a key modulator of plant nutrient use. However, evolutionary patterns for nutrient resorption remain unclear, especially in herbs. We measured nitrogen and phosphorus resorption on preselected leaves across the Helianthus (sunflower) genus in a common garden in Athens, GA. We analyzed our data with published leaf traits and native habitat environmental data. Using phylogenetically controlled analyses, we tested if (1) nutrient resorption correlates with leaf economic, vasculature, and defense traits through evolutionary time, and (2) native habitat environment predicts nutrient resorption evolution. For Helianthus, nutrient resorption capacity is greater in resource-conservative species, as previously defined for Helianthus with a principle components analysis of leaf economic spectrum traits (photosynthetic rate, respiration rate, leaf lifespan, leaf mass per area, and green-leaf N and P concentrations). Nutrient resorption capacity also evolutionarily correlates with individual leaf economic traits, though not always as expected based on broad species surveys. Greater nutrient resorption is also positively associated with leaf chemical defenses, but not leaf vasculature or senescence rate. Finally, nitrogen resorption evolution increases with native habitat precipitation, but native habitat soil fertility does not predict nitrogen or phosphorus resorption. Our results suggest nutrient resorption evolution is more closely tied to resource economic strategy than native habitat.

Background There is a growing appreciation that the evolution of floral traits is shaped by selection not only from pollinators, but also factors like abiotic stress, florivory, and disease. Many suc

Background and Aims Trait-based plant ecology attempts to use small numbers of functional traits to predict plant ecological strategies. However, a major gap exists between our understanding of organ-level ecophysiological traits and our understanding of whole-plant fitness and environmental adaptation. In this gap lie whole-plant organizational traits, including those that describe how plant biomass is allocated among organs and the timing of plant reproduction. This study explores the role of whole-plant organizational traits in adaptation to diverse environments in the context of life history, growth form and leaf economic strategy in a well-studied herbaceous system. Methods A phylogenetic comparative approach was used in conjunction with common garden phenotyping to assess the evolution of biomass allocation and reproductive timing across 83 populations of 27 species of the diverse genus Helianthus (the sunflowers). Key Results Broad diversity exists among species in both relative biomass allocation and reproductive timing. Early reproduction is strongly associated with resource-acquisitive leaf economic strategy, while biomass allocation is less integrated with either reproductive timing or leaf economics. Both biomass allocation and reproductive timing are strongly related to source site environmental characteristics, including length of the growing season, temperature, precipitation and soil fertility. Conclusions Herbaceous taxa can adapt to diverse environments in many ways, including modulation of phenology, plant architecture and organ-level ecophysiology. Although leaf economic strategy captures one key aspect of plant physiology, on their own leaf traits are not particularly predictive of ecological strategies in Helianthus outside of the context of growth form, life history and whole-plant organization. These results highlight the importance of including data on whole-plant organization alongside organ-level ecophysiological traits when attempting to bridge the gap between functional traits and plant fitness and environmental adaptation.

Recent work suggests variation in plant growth strategies is governed by a tradeoff in resource acquisition and use, ranging from a rapid resource acquisition strategy to a resource‐conservative strategy. While evidence for this tradeoff has been found in leaves, knowledge of root trait strategies, and whether they reflect adaptive differentiation across environments, is limited. In the greenhouse, we investigated variation in fine root morphology (specific root length and tissue density), chemistry (nitrogen concentration and carbon:nitrogen), and anatomy (root cross‐sectional traits) in populations of 26 Helianthus species and sister Phoebanthus tenuifolius. We also compared root trait variation in this study with leaf trait variation previously reported in a parallel study of these populations. Root traits varied widely and exhibited little phylogenetic signal, suggesting high evolutionary lability. Specific root length and root tissue density were weakly negatively correlated, but neither was associated with root nitrogen, providing little support for a single axis of root trait covariation. Correlations between traits measured in the greenhouse and native site characteristics were generally weak, suggesting a variety of equally viable root trait combinations exist within and across environments. However, high root nitrogen was associated with lower xylem vessel number and cross‐sectional area, suggesting a tradeoff between nutrient investment and water transport capacity. This led to correlations between root and leaf traits that were not always consistent with an acquisition–conservation tradeoff at the whole‐plant level. Given that roots must balance acquisition of water and nutrients with functions like anchorage, exudation, and microbial symbioses, the varied evidence for root trait covariation likely reflects the complexity of interacting selection pressures belowground. Similarly, the lack of evidence for a single acquisition–conservation tradeoff at the whole‐plant level likely reflects the vastly different selection pressures shaping roots and leaves, and the resources they are optimized to obtain.

Leaf defenses are widely recognized as key adaptations and drivers of plant evolution. Across environmentally diverse habitats, the macroevolution of leaf defenses can be predicted by the univariate trade-off model, which predicts that defenses are functionally redundant and thus trade off, and the resource availability hypothesis, which predicts that defense investment is determined by inherent growth rate and that higher defense will evolve in lower resource environments. Here, we examined the evolution of leaf physical and chemical defenses and secondary metabolites in relation to environmental characteristics and leaf economic strategy across 28 species of Helianthus (the sunflowers). Using a phylogenetic comparative approach, we found few evolutionary trade-offs among defenses and no evidence for defense syndromes. We also found that leaf defenses are strongly related to leaf economic strategy, with higher defense in more resource-conservative species, although there is little support for the evolution of higher defense in low-resource habitats. A wide variety of physical and chemical defenses predict resistance to different insect herbivores, fungal pathogens, and a parasitic plant, suggesting that most sunflower defenses are not redundant in function and that wild Helianthus represents a rich source of variation for the improvement of crop sunflower.

The leaf economics spectrum (LES) is a prominent ecophysiological paradigm that describes global variation in leaf physiology across plant ecological strategies using a handful of key traits. Nearly a decade ago, Shipley et al. (2006) used structural equation modelling to explore the causal functional relationships among LES traits that give rise to their strong global covariation. They concluded that an unmeasured trait drives LES covariation, sparking efforts to identify the latent physiological trait underlying the 'origin' of the LES. Here, we use newly developed phylogenetic structural equation modelling approaches to reassess these conclusions using both global LES data as well as data collected across scales in the genus Helianthus. For global LES data, accounting for phylogenetic non-independence indicates that no additional unmeasured traits are required to explain LES covariation. Across datasets in Helianthus, trait relationships are highly variable, indicating that global-scale models may poorly describe LES covariation at non-global scales.

The leaf economics spectrum (LES) describes a major axis of plant functional trait variation worldwide, defining suites of leaf traits aligned with resource-acquisitive to resource-conservative ecological strategies. The LES has been interpreted to arise from leaf-level trade-offs among ecophysiological traits common to all plants. However, it has been suggested that the defining leaf-level trade-offs of the LES may not hold within specific functional groups (e.g., herbs) nor within many groups of closely related species, which challenges the usefulness of the LES paradigm across evolutionary scales. Here, we examine the evolution of the LES across 28 species of the diverse herbaceous genus Helianthus (the sunflowers), which occupies a wide range of habitats and climate variation across North America. Using a phylogenetic comparative approach, we find repeated evolution of more resource-acquisitive LES strategies in cooler, drier, and more fertile environments. We also find macroevolutionary correlations among LES traits that recapitulate aspects of the global LES, but with one major difference: leaf mass per area is uncorrelated with leaf lifespan. This indicates that whole-plant processes likely drive variation in leaf lifespan across Helianthus, rather than leaf-level trade-offs. These results suggest that LES patterns do not reflect universal physiological trade-offs at small evolutionary scales.

Summary 1. Efforts to understand the effects of plant traits on carbon and nutrient cycling have recently focused on species variation and the potential for species data to improve predictions of past, present and future variation in ecosystems. However, the evolutionary lability of relevant traits among closely related species and the extent of intraspecific variation warrant further consideration. 2. Here, we examine interspecific and intraspecific variation in leaf N, LMA, root N and SRL at multiple scales, using Helianthus as a representative study system. 3. Substantial evolutionary lability of traits is demonstrated by interspecific variation in phylogenetically explicit analyses of closely related Helianthus species, and population differentiation of wild H. anomalus and cultivated H. annuus. 4. Intraspecific variation in leaf N and LMA, including genetic, environmental and ontogenetic responses, demonstrates that trait values for a single species can encompass a surprisingly large portion of the range encompassed by species in the leaf economics spectrum. 5. Synthesis. We recommend using data from selected natural populations to model effects of leaf N and LMA on decomposition, while using data from common garden experiments to determine evolutionary lability and thus inform potential for evolutionary change. If the high evolutionary lability of traits demonstrated for Helianthus is found for other important genera, this would suggest that these key ecophysiological traits are likely to respond to the selective pressures of global climate and land-use change.

• Plant resource-use traits are generally hypothesized to be adaptively differentiated for populations distributed along resource gradients. Although nutrient limitations are expected to select for resource-conservative strategies, water limitations may select for either resource-conservative or -acquisitive strategies. We test whether population differentiation reflects local adaptation for traits associated with resource-use strategies in a desert annual (Helianthus anomalus) distributed along a gradient of positively covarying water and nutrient availability. • We compared quantitative trait variation (Q(ST)) with neutral genetic differentiation (F(ST)), in a common garden glasshouse study, for leaf economics spectrum (LES) and related traits: photosynthesis (A(mass), A(area)), leaf nitrogen (N(mass), N(area)), leaf lifetime (LL), leaf mass per area (LMA), leaf water content (LWC), water-use efficiency (WUE, estimated as δ(13)C) and days to first flower (DFF). • Q(ST)-F(ST) differences support adaptive differentiation for Amass , N(mass), N(area), LWC and DFF. The trait combinations associated with drier and lower fertility sites represent correlated trait evolution consistent with the more resource-acquisitive end of the LES. There was no evidence for adaptive differentiation for A(area), LMA and WUE. • These results demonstrate that hot dry environments can selectively favor correlated evolution of traits contributing to a resource-acquisitive and earlier reproduction 'escape' strategy, despite lower fertility.

Granite outcrops in the southeastern United States are rare and isolated habitats that support edaphically controlled communities dominated by herbaceous plants. They harbor rare and endemic species that are expected to have low genetic variability and high population structure due to small population sizes and their disjunct habitat. We test this expectation for an annual outcrop endemic, Helianthus porteri (Porter's sunflower). Contrary to expectation, H. porteri has relatively high genetic diversity (H e = 0.681) and relatively low genetic structure among the native populations (F ST = 0.077) when compared to 5 other Helianthus species (N = 288; 18 expressed sequence tag-SSR markers). These findings suggest greater gene flow than expected. The potential for gene flow is supported by the analysis of transplant populations established with propagules from a common source in 1959. One population established close to a native population (1.5 km) at the edge of the natural range is genetically similar to and shares rare alleles with the adjacent native population and is distinct from the central source population. In contrast, a transplant population established north of the native range has remained similar to the source population. The relatively high genetic diversity and low population structure of this species, combined with the long-term success of transplanted populations, bode well for its persistence as long as the habitat persists.

Premise of the study Patterns of plant stem traits are expected to align with a "fast-slow" plant economic spectrum across taxa. Although broad patterns support such tradeoffs in field studies, tests of hypothesized correlated trait evolution and adaptive differentiation are more robust when taxa relatedness and environment are taken into consideration. Here we test for correlated evolution of stem and leaf traits and their adaptive differentiation across environments in the herbaceous genus, Helianthus. Methods Stem and leaf traits of 14 species of Helianthus (28 populations) were assessed in a common garden greenhouse study. Phylogenetically independent contrasts were used to test for evidence of correlated evolution of stem hydraulic and biomechanical properties, correlated evolution of stem and leaf traits, and adaptive differentiation associated with source habitat environments. Key results Among stem traits, there was evidence for correlated evolution of some hydraulic and biomechanical properties, supporting an expected tradeoff between stem theoretical hydraulic efficiency and resistance to bending stress. Population differentiation for suites of stem and leaf traits was found to be consistent with a "fast-slow" resource-use axis for traits related to water transport and use. Associations of population traits with source habitat characteristics supported repeated evolution of a resource-acquisitive "drought-escape" strategy in arid environments. Conclusions This study provides evidence of correlated evolution of stem and leaf traits consistent with the fast-slow spectrum of trait combinations related to water transport and use along the stem-to-leaf pathway. Correlations of traits with source habitat characteristics further indicate that the correlated evolution is associated, at least in part, with adaptive differentiation of Helianthus populations among native habitats differing in climate.

Plant roots exude numerous metabolites into the soil that influence nutrient availability. Although root exudate composition is hypothesized to be under selection in low fertility soils, few studies have tested this hypothesis in a phylogenetic framework. In this study, we examined root exudates of three pairs of Helianthus species chosen as phylogenetically-independent contrasts with respect to native soil nutrient availability. Under controlled environmental conditions, seedlings were grown to the three-leaf-pair stage, then transferred to either high or low nutrient treatments. After five days of nutrient treatments, we used gas chromatography-mass spectrometry for analysis of root exudates, and detected 37 metabolites across species. When compared in the high nutrient treatment, species native to low nutrient soils exhibited overall higher exudation than their sister species native to high nutrient soils in all three species pairs, providing support for repeated evolutionary shifts in response to native soil fertility. Species native to low nutrient soils and those native to high nutrient soils responded similarly to low nutrient treatments with increased exudation of organic acids (fumaric, citric, malic acids) and glucose, potentially as a mechanism to enhance nutrition acquisition. However, species native to low nutrient soils also responded to low nutrient treatments with a larger decrease in exudation of amino acids than species native to high nutrient soils in all three species pairs. This indicates that species native to low nutrient soils have evolved a unique sensitivity to changes in nutrient availability for some, but not all, root exudates. Overall, these repeated evolutionary divergences between species native to low nutrient soils and those native to high nutrient soils provide evidence for the adaptive value of root exudation, and its plasticity, in contrasting soil environments.

Assessing levels and patterns of population genetic variation is an important step for evaluating rare or endangered species and determining appropriate conservation strategies. This is particularly important for ensuring the preservation of novel genetic variation in wild relatives of crops, which could provide beneficial alleles for plant breeding and improvement. In this study, we evaluate the population genetics of Helianthus niveus ssp. tephrodes (the Algodones sunflower), which is an endangered, wild relative of cultivated sunflower (H. annuus L.). This rare sunflower species is native to the sand dunes of the Sonoran Desert in southern California, southwestern Arizona, and northern Mexico and is thought to harbor beneficial alleles for traits related to drought tolerance. We genotyped nine populations of this species with a set of simple-sequence repeat markers derived from expressed sequence tags (EST-SSRs) and investigated levels of genetic diversity and population structure, in H. niveus ssp. tephrodes. We also compared our results to findings from five related sunflower species that have been analyzed with these same markers, including annuals and perennials that range from rare to widespread. The Algodones sunflower harbors lower levels of standing genetic variation, but similar levels of population structure as compared to other sunflower species. We also discovered that a disjunct population from northern Mexico was genetically distinct from populations elsewhere in the range. Given the occurrence of such a genetically unique population, our recommendations include population surveys of the southern portion of the range in hopes of bolstering the existing germplasm collection.

Woody plants native to mesic habitats tend to be more vulnerable to drought-induced cavitation than those in xeric habitats. Cavitation resistance in herbaceous plants, however, is rarely studied and whether or not annual plants in arid habitats conform to the trends observed in woody plants is unknown. This question is addressed by comparing the hydraulic properties of annual plants endemic to relatively mesic and seasonally xeric habitats in the Great Basin Desert, in both native and experimental settings. Vulnerability to cavitation between species differed as predicted when vulnerability curves of similar-sized native individuals were compared. Contrary to expectations, Helianthus anomalus from the relatively mesic dune sites, on average, exhibited higher native embolism, lower soil-to-leaf hydraulic conductance (k(L)) and lower transpiration rates, than its xeric analogue, H. deserticola. In transplant gardens, H. anomalus' vulnerability to cavitation was unaffected by transplant location or watering treatment. In H. deserticola, however, vulnerability to cavitation varied significantly in response to watering in transplant gardens and varied as a function of stem water potential (Psi(stem)). H. deserticola largely avoided cavitation through its higher water status and generally more resistant xylem, traits consistent with a short life cycle and typical drought-escape strategy. By contrast, H. anomalus' higher native embolism is likely to be adaptive by lowering plant conductance and transpiration rate, thus preventing the loss of root-to-soil hydraulic contact in the coarse sand dune soils. For H. anomalus this dehydration avoidance strategy is consistent with its relatively long 3-4 month life cycle and low-competition habitat. We conclude that variance of hydraulic parameters in herbaceous plants is a function of soil moisture heterogeneity and is consistent with the notion that trait plasticity to fine-grained environmental variation can be adaptive.

C(3) plants dominate many landscapes and are critically important for ecosystem water cycling. At night, plant water losses can include transpiration (E(night)) from the canopy and hydraulic redistribution (HR) from roots. We tested whether E(night) limits the magnitude of HR in a greenhouse study using Artemisia tridentata, Helianthus anomalus and Quercus laevis. Plants were grown with their roots split between two compartments. HR was initiated by briefly withholding all water, followed by watering only one rooting compartment. Under study conditions, all species showed substantial E(night) and HR (highest minus lowest soil water potential [Psi(s)] during a specified diel period). Suppressing E(night) by canopy bagging increased HR during the nightly bagging period (HR(N)) for A. tridentata and H. anomalus by 73 and 33% respectively, but did not affect HR(N) by Q. laevis. Total daily HR (HR(T)) was positively correlated with the Psi(s) gradient between the rooting compartments, which was correlated with light and/or atmospheric vapour pressure deficit (VPDa) the prior day. For A. tridentata, HR(T) was negatively correlated with night-time VPDa. Ecological implications of the impact of E(night) on HR may include decreased plant productivity during dry seasons, altered ecosystem water flux patterns and reduced nutrient cycling in drying soils.

Habitats that differ in soil resource availability are expected to differ for selection on resource-related plant traits. Here, we examined spatial and temporal variation in phenotypic selection on leaf ecophysiological traits for 10 Helianthus populations, including two species of hybrid origin, Helianthus anomalus and Helianthus deserticola, and artificial hybrids of their ancestral parents. Leaf traits assessed were leaf size, succulence, nitrogen (N) concentration and water-use efficiency (WUE). Biomass and leaf traits of artificial hybrids indicate that the actively moving dune habitat of H. anomalus was more growth limiting, with lower N availability but higher relative water availability than the stabilized dune habitat of H. deserticola. Habitats differed for direct selection on leaf N and WUE, but not size or succulence, for the artificial hybrids. However, within the H. anomalus habitat, direct selection on WUE also differed among populations. Across years, direct selection on leaf traits did not differ. Leaf N was the only trait for which direct selection differed between habitats but not within the H. anomalus habitat, suggesting that nutrient limitation is an important selective force driving adaptation of H. anomalus to the active dune habitat.

Natural hybridization accompanied by a shift in niche preference by hybrid genotypes can lead to hybrid speciation. Natural selection may cause the fixation of advantageous alleles in the ecologically diverged hybrids, and the loci experiencing selection should exhibit a reduction in allelic diversity relative to neutral loci. Here, we analyzed patterns of genetic diversity at 59 microsatellite loci associated with expressed sequence tags (ESTs) in a homoploid hybrid sunflower species, Helianthus anomalus. We used two indices, ln RV and ln RH, to compare variation and heterozygosity (respectively) at each locus between the hybrid species and its two parental species, H. annuus and H. petiolaris. Mean values of ln RV and ln RH were significantly lower than zero, which implies that H. anomalus experienced a population bottleneck during its recent evolutionary history. After correcting for the apparent bottleneck, we found six loci with a significant reduction in variation or with heterozygosity in the hybrid species, compared to one or both of the parental species. These loci should be viewed as a ranked list of candidate loci, pending further sequencing and functional analyses. Sequence data were generated for two of the candidate loci, but population genetics tests failed to detect deviations from neutral evolution at either locus. Nonetheless, a greater than eight-fold excess of nonsynonymous substitutions was found near a putative N-myristoylation motif at the second locus (HT998), and likelihood-based models indicated that the protein has been under selection in H. anomalus in the past and, perhaps, in one or both parental species. Finally, our data suggest that selective sweeps may have united populations of H. anomalus isolated by a mountain range, indicating that even low gene-flow species may be held together by the spread of advantageous alleles.

The leaf economics spectrum (LES) describes a major axis of plant functional trait variation worldwide, defining suites of leaf traits aligned with resource-acquisitive to resource-conservative ecological strategies. The LES has been interpreted to arise from leaf-level trade-offs among ecophysiological traits common to all plants. However, it has been suggested that the defining leaf-level trade-offs of the LES may not hold within specific functional groups (e.g., herbs) nor within many groups of closely related species, which challenges the usefulness of the LES paradigm across evolutionary scales. Here, we examine the evolution of the LES across 28 species of the diverse herbaceous genus Helianthus (the sunflowers), which occupies a wide range of habitats and climate variation across North America. Using a phylogenetic comparative approach, we find repeated evolution of more resource-acquisitive LES strategies in cooler, drier, and more fertile environments. We also find macroevolutionary correlations among LES traits that recapitulate aspects of the global LES, but with one major difference: leaf mass per area is uncorrelated with leaf lifespan. This indicates that whole-plant processes likely drive variation in leaf lifespan across Helianthus, rather than leaf-level trade-offs. These results suggest that LES patterns do not reflect universal physiological trade-offs at small evolutionary scales.

We investigated the response of Helianthus species nighttime conductance (g night) and transpiration (E night) to soil nutrient and water limitations in nine greenhouse studies. The studies primarily used wild Helianthus annuus, but also included a commercial and early domesticate of H. annuus and three additional wild species (Helianthus petiolaris Nutt., Helianthus deserticola Heiser, and Helianthus anomalus Blake). Well-watered plants of all species showed substantial g night (0.023–0.225 mol m−2 s−1) and E night (0.29–2.46 mmol m−2 s−1) measured as instantaneous gas exchange. Based on the potential for transpiration to increase mass flow of mobile nutrients to roots, we hypothesized that g night and E night would increase under limiting soil nutrients but found no evidence of responses in all six studies testing this. Based on known daytime responses to water limitation, we hypothesized that g night and E night would decrease when soil water availability was limited, and results from all four studies testing this supported our hypothesis. We also established that stomatal conductance at night was on average 5 times greater than cuticular conductance. Additionally, g night and E night varied nocturnally and across plant reproductive stages while remaining relatively constant as leaves aged. Our results further the ability to predict conditions under which nighttime water loss will be biologically significant and demonstrate that for Helianthus, g night can be regulated.

We examined how plant traits related to growth and resource use have evolved during hybrid speciation and specialization into stressful habitats. Two desert sunflower species of homoploid hybrid origin are endemic to habitats with lower soil nutrient levels than those of their ancestral parent species. We hypothesized that the hybrid species would exhibit greater tolerance to low levels of soil nutrients than their parental species. The 2 hybrid species, Helianthus anomalus and H. deserticola, and their parental species, H. annuus and H. petiolaris, were compared for plant traits and growth through reproduction under 3 nutrient levels in a greenhouse study. An additional seedling study compared species for maximum seedling relative growth rate under optimum conditions. The hybrid species did have greater tolerance of nutrient limitation than the parental species, demonstrated by a resistance to change in stem height and diameter growth across treatments. A similar trend was observed in total biom...

Can the complex phenotypes that characterize naturally occurring hybrid species be re-created in early-generation artificial hybrids? We address this question with three homoploid hybrid species (Helianthus anomalus, Helianthus deserticola, Helianthus paradoxus) and their ancestral parents (Helianthus annuus, Helianthus petiolaris) that are phenotypically distinct and ecologically differentiated. These species, and two synthetic hybrid populations of the ancestral parents, were characterized for morphological, physiological, and life-history traits in greenhouse studies. Among the synthetic hybrids, discriminant analysis identified a few individuals with the multitrait phenotype of the natural hybrid species: 0.7%–1.1% were H. anomalus-like, 0.5%–13% were H. deserticola-like, and only 0.4% were H. paradoxus-like. These relative frequencies mirror previous findings that genetic correlations are favorable for generating the hybrid species’ phenotypes, and they correspond well with phylogeographic evidence that demonstrates multiple natural origins of H. deserticola and H. anomalus but a single origin for H. paradoxus. Even though synthetic hybrids with hybrid species phenotypes are rare, their phenotypic correlation matrices share most of the same principal components (eigenvectors), setting the stage for predictable recovery of hybrid species’ phenotypes from different hybrid populations. Our results demonstrate past hybridization could have generated hybrid species-like multitrait phenotypes suitable for persistence in their respective environments in just three generations after initial hybridization.

The diploid hybrid species Helianthus deserticola inhabits the desert floor, an extreme environment relative to its parental species Helianthus annuus and Helianthus petiolaris. Adaptation to the desert floor may have occurred via selection acting on transgressive, or extreme, traits in early hybrids between the parental species. We explored this possibility through a field experiment in the hybrid species' native habitat using H. deserticola, H. annuus, H. petiolaris, and two populations of early-generation (BC(2)) hybrids between the parental species, which served as proxies for the ancestral genotype of the ancient hybrid species. Character expression was evaluated for each genotypic class. Helianthus deserticola was negatively transgressive for stem diameter, leaf area, and flowering date, and the latter two traits are likely to be advantageous in a desert environment. The BC(2) hybrids contained a range of variation that overlapped these transgressive trait means, and an analysis of phenotypic selection revealed that some of the selective pressures on leaf size and flowering date, but not stem diameter, would move the BC(2) population toward the H. deserticola phenotype. Thus, H. deserticola may have originated from habitat-mediated directional selection acting on hybrids between H. annuus and H. petiolaris in a desert environment.

Hybridization is frequent in many organismal groups, but its role in adaptation is poorly understood. In sunflowers, species found in the most extreme habitats are ancient hybrids, and new gene combinations generated by hybridization are speculated to have contributed to ecological divergence. This possibility was tested through phenotypic and genomic comparisons of ancient and synthetic hybrids. Most trait differences in ancient hybrids could be recreated by complementary gene action in synthetic hybrids and were favored by selection. The same combinations of parental chromosomal segments required to generate extreme phenotypes in synthetic hybrids also occurred in ancient hybrids. Thus, hybridization facilitated ecological divergence in sunflowers.

The primary requirement for a new diploid species to arise via hybridization is ecological divergence from its parental species. Ecological divergence protects the nascent hybrid species from competition with its progenitor species and may contribute to reproductive isolation. However, the means by which hybridization might facilitate the necessary adaptive transitions are poorly understood. Here, we report the results of a glasshouse experiment in which 42 morphological and ecophysiological traits were measured in three hybrid sunflower species (Helianthus anomalus, Helianthus deserticola, and Helianthus paradoxus) and their parental species (Helianthus annuus and Helianthus petiolaris). A surprisingly high proportion of traits were extreme relative to the parental species (24%, 20%, and 39% of traits in H. anomalus, H. deserticola, and H. paradoxus, respectively). Most of the extreme traits have previously been reported in the literature as adaptations to dune (H. anomalus), high‐desert (H. deserticola)...

Leaf defenses have long been studied in the context of plant growth rate, resource availability, and optimal investment theory. Likewise, one of the central modern paradigms of plant ecophysiology, the leaf economics spectrum (LES), has been extensively studied in the context of these factors across ecological scales ranging from global species data sets to temporal shifts within individuals. Despite strong physiological links between LES strategy and leaf defenses in structure, function, and resource investment, the relationship between these trait classes has not been well explored. This study investigates the relationship between leaf defenses and LES strategy across whole-plant ontogeny in three diverse Helianthus species known to exhibit dramatic ontogenetic shifts in LES strategy, focusing primarily on physical and quantitative chemical defenses. Plants were grown under controlled environmental conditions and sampled for LES and defense traits at four ontogenetic stages. Defenses were found to shift strongly with ontogeny, and to correlate strongly with LES strategy. More advanced ontogenetic stages with more conservative LES strategy leaves had higher tannin activity and toughness in all species, and higher leaf dry matter content in two of three species. Modeling results in two species support the conclusion that changes in defenses drive changes in LES strategy through ontogeny, and in one species that changes in defenses and LES strategy are likely independently driven by ontogeny. Results of this study support the hypothesis that leaf-level allocation to defenses might be an important determinant of leaf economic traits, where high investment in defenses drives a conservative LES strategy.

Diploid hybrid lineages often are ecologically distinct from their parental species. However, it is unclear whether this niche divergence is typically achieved via hybrid intermediacy, a mixture of parental traits, and/or the evolution of extreme (transgressive) morphological and ecophysiological features. Here we compare an extensively studied hybrid sunflower species, Helianthus anomalus, with its putative parents, H. annuus and H. petiolaris, for 41 morphological and 12 ecophysiological traits. Helianthus anomalus was morphologically intermediate for one trait (2.4%), parental-like for 23 traits (56.1%), and transgressive for 17 traits (41.5%). For ecophysiological traits, H. anomalus was not significantly different from one or both parents for nine traits (75%), and was transgressive for the remaining three (25%). Thus, H. anomalus appears to be a mosaic of parental-like and transgressive phenotypes. Although the fitness effects of the transgressive characters are not yet known, many of these characters are consistent with adaptations reported for other sand dune plants. Genetic studies are currently underway to ascertain whether these extreme characters arose as a direct byproduct of hybridization or whether they evolved via mutational divergence.

The leaf economics spectrum (LES) describes large cross-species variation in suites of leaf functional traits ranging from resource-acquisitive to resource-conservative strategies. Such strategies have been integral in explaining plant adaptation to diverse environments, and have been linked to numerous ecosystem processes. The LES has previously been found to be significantly modulated by climate, soil fertility, biogeography, growth form, and life history. One largely unexplored aspect of LES variation, whole-plant ontogeny, is investigated here using multiple populations of three very different species of sunflower: Helianthus annuus, Helianthus mollis, and Helianthus radula. Plants were grown under environmentally controlled conditions and assessed for LES and related traits at four key developmental stages, using recently matured leaves to standardize for leaf age. Nearly every trait exhibited a significant ontogenetic shift in one or more species, with trait patterns differing among populations and species. Photosynthetic rate, leaf nitrogen concentration, and leaf mass per area exhibited surprisingly large changes, spanning over two-thirds of the original cross-species LES variation and shifting from resource-acquisitive to resource-conservative strategies as the plants matured. Other traits being investigated in relation to the LES, such as leaf water content, pH, and vein density, also showed large changes. The finding that ontogenetic variation in LES strategy can be substantial leads to a recommendation of standardization by developmental stage when assessing ‘species values’ of labile traits for comparative approaches. Additionally, the substantial ontogenetic trait shifts seen within single individuals provide an opportunity to uncover the contribution of gene regulatory changes to variation in LES traits.

Background and aims Low soil fertility limits growth and productivity in many natural and agricultural systems, where the ability to sense and respond to nutrient limitation is important for success. Helianthus anomalus is an annual sunflower of hybrid origin that is adapted to desert sand-dune substrates with lower fertility than its parental species, H. annuus and H. petiolaris. Previous studies have shown that H. anomalus has traits generally associated with adaptation to low-fertility habitats, including a lower inherent relative growth rate and longer leaf lifetime. Methods Here, a cDNA microarray is used to identify gene expression differences that potentially contribute to increased tolerance of low fertility of the hybrid species by comparing the nitrogen stress response of all three species with high- and low-nutrient treatments. Key results Relative to the set of genes on the microarray, the genes showing differential expression in the hybrid species compared with its parents are enriched in stress-response genes, developmental genes, and genes involved in responses to biotic or abiotic stimuli. After a correction for multiple comparisons, five unique genes show a significantly different response to nitrogen limitation in H. anomalus compared with H. petiolaris and H. annuus. The Arabidopsis thaliana homologue of one of the five genes, catalase 1, has been shown to affect the timing of leaf senescence, and thus leaf lifespan. Conclusions The five genes identified in this analysis will be examined further as candidate genes for the adaptive stress response in H. anomalus. Genes that improve growth and productivity under nutrient stress could be used to improve crops for lower soil fertility which is common in marginal agricultural settings.

Studies of the genetic architecture of ecologically important traits shed light on the evolution of those traits in natural populations. Studies of genetic architecture of complex traits are important because they can predict whether a phenotypic transition involved major leaps or occurred more smoothly (Burke et al. 2002). Also, alleles with large effects will fix more rapidly than will those of very small effect (Barton and Keightley 2002). An historical illustration of genetic architecture is the study by Beadle (1972) examining the inheritance of maize domestication traits. Using a primitive landrace of maize and a teosinte accession, Beadle observed the segregation of the maize and teosinte phenotypes at a 1:500 ratio, indicating that 4 or 5 genes of major effect were responsible for the bulk of changes from teosinte to maize (Beadle 1972). This demonstrated that the phenotypic transition from wild progenitor to primitive maize likely occurred in major leaps. This contrasts with sunflower, where domestication appears to have involved many small quantitative trait loci (QTLs), suggesting a smoother and more gradual transition to the domesticated form (Burke et al. 2002). Phenotypic differences between wild sunflower species (Helianthus) also seem to be controlled by a large number of loci, each with a relatively small contribution to the phenotype (Kim and Rieseberg 1999; Rieseberg et al. 2003; Lexer et al. 2005). Two Helianthus species that have been the targets of previous genetic study, Helianthus annuus and Helianthus petiolaris, are of particular interest because they hybridized to form 3 stable, ancient, diploid hybrid species Helianthus anomalus, Helianthus deserticola, and Helianthus paradoxus (Rieseberg et al. 1990; Rieseberg 1991), which occupy extreme habitats compared with the parents: active sand dunes, dry desert floors, and brackish salt marshes, respectively. The range of phenotypes observed in the hybrid species is also more extreme than those of the parental species (Schwarzbach et al. 2001; Rosenthal et al. 2002). Evolution of the hybrid sunflower species appears to have occurred quickly (Buerkle CA and Rieseberg LH, in review; Ungerer et al. 1998), and 2 of the hybrid species may have multiple origins (H. anomalus, Schwarzbach and Rieseberg 2002 and H. deserticola, Gross et al. 2003). Additionally, phenotypes of the stable hybrid species are present in populations of early hybrids between H. annuus and H. petiolaris (Rosenthal, Rieseberg, and Donovan 2005). If the preadaptive phenotypic differences important for colonizing the extreme habitats of the hybrid species are controlled by multiple genes of small effect, then the rate of fixation of individual QTL alleles is likely to be slow and the sizes of parental chromosomal blocks in the hybrid species should be smaller than when loci of large effect control the phenotype (Barton and Keightley 2002). It seems paradoxical that most of the QTLs in Helianthus tend to be of small effect, but the hybrid species genomes seem to have stabilized relatively quickly (Buerkle and Rieseberg, in review; Ungerer et al. 1998). It may be the case, however, that variation in some of the traits that are important to the survival of early-generation hybrids in Helianthus are controlled by large-effect QTLs. Also, we know that pollen sterility QTLs, which also affect hybrid genomic composition (Rieseberg et al. 1996; Karrenberg et al. forthcoming), are controlled by major QTLs (Lai et al. 2005). Here, we examine leaf ecophysiological traits, some of which greatly affect fitness of plants in the wild. Leaf nitrogen is likely to be ecologically important because it roughly estimates investment in nitrogen-rich photosynthetic enzymes, with investment per unit leaf area influencing photosynthetic capacity (Field and Mooney 1986). Foliar nitrogen is positively transgressive in the hybrid sunflower species H. anomalus (Rosenthal et al. 2002) and has been shown to impact survival in its native habitat (Ludwig et al. 2004). Despite being an important physiological trait, leaf nitrogen has only recently received attention in QTL analyses (Ishimaru et al. 2001; Hall et al. 2005; Takai et al. 2006; Weih et al. 2006). Other leaf chemistry traits measured in this study assess important aspects of plant physiology that are related to nitrogen allocation. Leaf carbon is a rough assessment of investment in carbon-rich structural molecules such as cellulose and correlates with construction cost of leaves (Nagel et al. 2002). Carbon isotopic ratio (δ13C) is often used as a proxy for water-use efficiency in C3 plants (Farquhar et al. 1989; Ehleringer et al. 1992). Differences in leaf nitrogen isotopic composition (δ15N) for plants grown under similar conditions imply differences in nitrogen uptake or use (Evans 2001). Photosynthetic nitrogen use efficiency (PNUE) is the instantaneous rate of photosynthesis per unit leaf nitrogen and may be under selection in nutrient-poor or water-limited habitats (Field and Mooney 1986; Fredeen et al. 1991; Wright et al. 2002). Here we report the results of QTL analyses for these ecologically important, physiologically based leaf traits.

Plant water-use efficiency (WUE) is expected to affect plant fitness and thus be under natural selection in arid habitats. Although many natural population studies have assessed plant WUE, only a few related WUE to fitness. The further determination of whether selection on WUE is direct or indirect through functionally related traits has yielded no consistent results. For natural populations of two desert annual sunflowers, Helianthus anomalus and H. deserticola, we used phenotypic selection analysis with vegetative biomass as the proxy for fitness to test (1) whether there was direct and indirect selection on WUE (carbon isotope ratio) and related traits (leaf N, area, succulence) and (2) whether direct selection was consistent with hypothesized drought/dehydration escape and avoidance strategies. There was direct selection for lower WUE in mesic and dry H. anomalus populations, consistent with dehydration escape, even though it is the longer lived of the two species. For mesic H. anomalus, direct selection favored lower WUE and higher N, suggesting that plants may be "wasting water" to increase N delivery via the transpiration stream. For the shorter lived H. deserticola in the direr habitat, there was indirect selection for lower WUE, inconsistent with drought escape. There was also direct selection for higher leaf N, succulence and leaf size. There was no direct selection for higher WUE consistent with dehydration avoidance in either species. Thus, in these natural populations of two desert dune species higher fitness was associated with some combination direct and indirect selection for lower WUE, higher leaf N and larger leaf size. Our understanding of the adaptive value of plant ecophysiological traits will benefit from further consideration of related traits such as leaf nitrogen and more tests in natural populations.

Recent research has shown that many C3 plant species have significant stomatal opening and transpire water at night even in desert habitats. Day-time stomatal regulation is expected to maximize carbon gain and prevent runaway cavitation, but little is known about the effect of soil resource availability on night-time stomatal conductance (g) and transpiration (E). Water (low and high) and nutrients (low and high) were applied factorially during the growing season to naturally occurring seedlings of the annual Helianthus anomalus. Plant height and biomass were greatest in the treatment where both water and nutrients were added, confirming resource limitations in this habitat. Plants from all treatments showed significant night-time g (approximately 0.07 mol m(-2) s(-1)) and E (approximately 1.5 mol m(-2) s(-1)). In July, water and nutrient additions had few effects on day- or night-time gas exchange. In August, however, plants in the nutrient addition treatments had lower day-time photosynthesis, g and E, paralleled by lower night-time g and E. Lower predawn water potentials and higher integrated photosynthetic water-use efficiency suggests that the nutrient addition indirectly induced a mild water stress. Thus, soil resources can affect night-time g and E in a manner parallel to day-time, although additional factors may also be involved.

Much of our knowledge of speciation genetics stems from quantitative trait locus (QTL) studies. However, interpretations of the size and distribution of QTL underlying species differences are complicated by differences in the way QTL magnitudes are estimated. Also, many studies fail to exploit information about QTL directions or to compare inter- and intraspecific QTL variation. Here, we comprehensively analyze an extensive QTL data set for an interspecific backcross between two wild annual sunflowers, Helianthus annuus and H. petiolaris, interpret different estimates of QTL magnitudes, identify trait groups that have diverged through selection, and compare inter- and intraspecific QTL magnitudes. Our results indicate that even minor QTL (in terms of backcross variance) may be surprisingly large compared to levels of standing variation in the parental species or phenotypic differences between them. Morphological traits, particularly flower morphology, were more strongly or consistently selected than life history or physiological traits. Also, intraspecific QTL were generally smaller than interspecific ones, consistent with the prediction that larger QTL are more likely to spread to fixation across a subdivided population. Our results inform the genetics of species differences in Helianthus and suggest an approach for the simultaneous mapping of inter- and intraspecific QTL.