Adaptive genetic potential and plasticity of trait variation in the foundation prairie grass Andropogon gerardii across the US Great Plains' climate gradient: Implications for climate change and restoration.

Plant response to climate depends on a species' adaptive potential. To address this, we used reciprocal gardens to detect genetic and environmental plasticity effects on phenotypic variation and combined with genetic analyses. Four reciprocal garden sites were planted with three regional ecotypes of Andropogon gerardii, a dominant Great Plains prairie grass, using dry, mesic, and wet ecotypes originating from western KS to Illinois that span 500-1,200 mm rainfall/year. We aimed to answer: (a) What is the relative role of genetic constraints and phenotypic plasticity in controlling phenotypes? (b) When planted in the homesite, is there a trait syndrome for each ecotype? (c) How are genotypes and phenotypes structured by climate? and (d) What are implications of these results for response to climate change and use of ecotypes for restoration? Surprisingly, we did not detect consistent local adaptation. Rather, we detected co-gradient variation primarily for most vegetative responses. All ecotypes were stunted in western KS. Eastward, the wet ecotype was increasingly robust relative to other ecotypes. In contrast, fitness showed evidence for local adaptation in wet and dry ecotypes with wet and mesic ecotypes producing little seed in western KS. Earlier flowering time in the dry ecotype suggests adaptation to end of season drought. Considering ecotype traits in homesite, the dry ecotype was characterized by reduced canopy area and diameter, short plants, and low vegetative biomass and putatively adapted to water limitation. The wet ecotype was robust, tall with high biomass, and wide leaves putatively adapted for the highly competitive, light-limited Eastern Great Plains. Ecotype differentiation was supported by random forest classification and PCA. We detected genetic differentiation and outlier genes associated with primarily precipitation. We identified candidate gene GA1 for which allele frequency associated with plant height. Sourcing of climate adapted ecotypes should be considered for restoration.

Local adaptation, genetic divergence, and experimental selection in a foundation grass across the US Great Plains' climate gradient.

Many prior studies have uncovered evidence for local adaptation using reciprocal transplant experiments. However, these studies are rarely conducted for a long enough time to observe succession and competitive dynamics in a community context, limiting inferences for long-lived species. Furthermore, the genetic basis of local adaptation and genetic associations with climate has rarely been identified. Here, we report on a long-term (6-year) experiment conducted under natural conditions focused on Andropogon gerardii, the dominant grass of the North American Great Plains tallgrass ecosystem. We focus on this foundation grass that comprises 80% of tallgrass prairie biomass and is widely used in 20,000 km2 of restoration. Specifically, we asked the following questions: (a) Whether ecotypes are locally adapted to regional climate in realistic ecological communities. (b) Does adaptive genetic variation underpin divergent phenotypes across the climate gradient? (c) Is there evidence of local adaptation if the plants are exposed to competition among ecotypes in mixed ecotype plots? Finally, (d) are local adaptation and genetic divergence related to climate? Reciprocal gardens were planted with 3 regional ecotypes (originating from dry, mesic, wet climate sources) of Andropogon gerardii across a precipitation gradient (500-1,200 mm/year) in the US Great Plains. We demonstrate local adaptation and differentiation of ecotypes in wet and dry environments. Surprisingly, the apparent generalist mesic ecotype performed comparably under all rainfall conditions. Ecotype performance was underpinned by differences in neutral diversity and candidate genes corroborating strong differences among ecotypes. Ecotype differentiation was related to climate, primarily rainfall. Without long-term studies, wrong conclusions would have been reached based on the first two years. Further, restoring prairies with climate-matched ecotypes is critical to future ecology, conservation, and sustainability under climate change.

The role of ecotypic variation and the environment on biomass and nitrogen in a dominant prairie grass.

Knowledge of the relative strength of evolution and the environment on a phenotype is required to predict species responses to environmental change and decide where to source plant material for ecological restoration. This information is critically needed for dominant species that largely determine the productivity of the central U.S. grassland. We established a reciprocal common garden experiment across a longitudinal gradient to test whether ecotypic variation interacts with the environment to affect growth and nitrogen (N) storage in a dominant grass. We predicted plant growth would increase from west to east, corresponding with increasing precipitation, but differentially among ecotypes due to local adaptation in all ecotypes and a greater range of growth response in ecotypes originating from west to east. We quantified aboveground biomass, root biomass, belowground net primary production (BNPP), root C:N ratio, and N storage in roots of three ecotypes of Andropogon gerardii collected from and reciprocally planted in central Kansas, eastern Kansas, and s6uthern Illinois. Only the ecotype from the most mesic region (southern Illinois) exhibited more growth from west to east. There was evidence for local adaptation in the southern Illinois ecotype by means of the local vs. foreign contrast within a site and the home vs. away contrast when growth in southern Illinois was compared to the most distant 'site in central Kansas. Root biomass of the eastern Kansas ecotype was higher at home than at either away site. The ecotype from the driest region, central Kansas, exhibited the least response across the environmental gradient, resulting in a positive relationship between the range of biomass response and precipitation in ecotype region of origin. Across all sites, ecotypes varied in root C:N ratio (highest in the driest-origin ecotype) and N storage in roots (highest in the most mesic-origin ecotype). The low and limited range of biomass, higher C:N ratio of roots, and lower N storage in the central Kansas ecotype relative to the southern Illinois ecotype suggests that introducing ecotypes of A. gerardii from much drier regions into highly mesic prairie would reduce productivity and alter belowground ecosystem processes under a wide range of conditions.

Intraspecific variation of a dominant grass and local adaptation in reciprocal garden communities along a US Great Plains' precipitation gradient: implications for grassland restoration with climate change.

Identifying suitable genetic stock for restoration often employs a 'best guess' approach. Without adaptive variation studies, restoration may be misguided. We test the extent to which climate in central US grasslands exerts selection pressure on a foundation grass big bluestem (Andropogon gerardii), widely used in restorations, and resulting in local adaptation. We seeded three regional ecotypes of A. gerardii in reciprocal transplant garden communities across 1150 km precipitation gradient. We measured ecological responses over several timescales (instantaneous gas exchange, medium-term chlorophyll absorbance, and long-term responses of establishment and cover) in response to climate and biotic factors and tested if ecotypes could expand range. The ecotype from the driest region exhibited greatest cover under low rainfall, suggesting local adaptation under abiotic stress. Unexpectedly, no evidence for cover differences between ecotypes exists at mesic sites where establishment and cover of all ecotypes were low, perhaps due to strong biotic pressures. Expression of adaptive differences is strongly environment specific. Given observed adaptive variation, the most conservative restoration strategy would be to plant the local ecotype, especially in drier locations. With superior performance of the most xeric ecotype under dry conditions and predicted drought, this ecotype may migrate eastward, naturally or with assistance in restorations.

Species-specific enzymatic tolerance of sulfide toxicity in plant roots.

Toxic effects of sulfide come from a poisoning of a number of enzymes, especially cytochrome c oxidase, which catalyzes the terminal step in mitochondrial aerobic respiration. Despite this, some estuarine plants live in sulfide-rich sediments. We hypothesized estuarine and flooding-tolerant species might be more tolerant of sulfide compared to upland species, and this was tested by measures of root cytochrome c oxidase and alcohol dehydrogenase activities in extracts exposed to sulfide. Enzyme activities were measured in 0, 5, 10, 15, and 20 µM sodium sulfide, and compared among 17 species of plants. Activities of alcohol dehydrogenase and cytochrome c oxidase were both reduced by increasing sulfide concentration, but cytochrome c oxidase was more sensitive to sulfide compared to alcohol dehydrogenase. Activities of cytochrome c oxidase were reduced to near zero at 5-10 µM sulfide whereas alcohol dehydrogenase activities were only reduced by about 50% at 10 µM sulfide. All species were sensitive to increasing sulfide, but to different degrees. Cytochrome c oxidase in flooding-sensitive species was decreased to near zero activity at 5 µM sulfide, whereas activities in some flooding-tolerant species were still detectable until 15 µM sulfide. Cytochrome c oxidase activities in some estuarine species were low even in the absence of sulfide, perhaps an adaptation to avoid sulfide vulnerability in their native, sulfide-rich habitat. This illustrates the potent metabolic effects of sulfide, and this is the first demonstration of varying sensitivities of cytochrome c oxidase to sulfide across organisms, making these data of novel importance.

Ecotypes of an ecologically dominant prairie grass (Andropogon gerardii) exhibit genetic divergence across the U.S. Midwest grasslands' environmental gradient.

Big bluestem (Andropogon gerardii) is an ecologically dominant grass with wide distribution across the environmental gradient of U.S. Midwest grasslands. This system offers an ideal natural laboratory to study population divergence and adaptation in spatially varying climates. Objectives were to: (i) characterize neutral genetic diversity and structure within and among three regional ecotypes derived from 11 prairies across the U.S. Midwest environmental gradient, (ii) distinguish between the relative roles of isolation by distance (IBD) vs. isolation by environment (IBE) on ecotype divergence, (iii) identify outlier loci under selection and (iv) assess the association between outlier loci and climate. Using two primer sets, we genotyped 378 plants at 384 polymorphic AFLP loci across regional ecotypes from central and eastern Kansas and Illinois. Neighbour-joining tree and PCoA revealed strong genetic differentiation between Kansas and Illinois ecotypes, which was better explained by IBE than IBD. We found high genetic variability within prairies (80%) and even fragmented Illinois prairies, surprisingly, contained high within-prairie genetic diversity (92%). Using Bayenv2, 14 top-ranked outlier loci among ecotypes were associated with temperature and precipitation variables. Six of seven BayeScanFST outliers were in common with Bayenv2 outliers. High genetic diversity may enable big bluestem populations to better withstand changing climates; however, population divergence supports the use of local ecotypes in grassland restoration. Knowledge of genetic variation in this ecological dominant and other grassland species will be critical to understanding grassland response and restoration challenges in the face of a changing climate.

Environmental and genetic variation in leaf anatomy among populations of Andropogon gerardii (Poaceae) along a precipitation gradient.

PREMISE OF THE STUDY: Phenotypes of two Andropogon gerardii subspecies, big bluestem and sand bluestem, vary throughout the prairie ecosystem of North America. This study sought to determine the role of genetics and environment in driving adaptive variation of leaf structure in big bluestem and sand bluestem. • METHODS: Four populations of big bluestem and one population of sand bluestem were planted in common gardens at four sites across a precipitation gradient from western Kansas to southern Illinois. Internal leaf structure and trichome density of A. gerardii were examined by light microscopy to separate genetic and environmentally controlled traits. Leaf thickness, midrib thickness, bulliform cells, interveinal distance, vein size, and trichome density were quantified. • KEY RESULTS: At all planting sites, sand bluestem and the xeric population of A. gerardii had thicker leaves and fewer bulliform cells compared with mesic populations. Environment and genetic source population were both influential for leaf anatomy. Leaves from plants grown in mesic sites (Carbondale, Illinois and Manhattan, Kansas) had thicker midribs, larger veins, fewer trichomes, and a greater proportion of bulliform cells compared to plants grown in drier sites (Colby and Hays, Kansas). • CONCLUSIONS: Water availability has driven adaptive variation in leaf structure in populations of A. gerardii, particularly between sand bluestem and big bluestem. Genetically based differences in leaves of A. gerardii indicate adaptive variation and evolutionary forces differentiating sand bluestem from big bluestem. Environmental responses of A. gerardii leaves suggest an ability to adjust to drought, even in populations adapted to mesic home environments.

Diversity in leaf anatomy, and stomatal distribution and conductance, between salt marsh and freshwater species in the C(4) genus Spartina (Poaceae).

Leaf anatomy, stomatal density, and leaf conductance were studied in 10 species of Spartina (Poaceae) from low versus high salt marsh, and freshwater habitats. Internal structure, external morphology, cuticle structure, and stomatal densities were studied with light and electron microscopy. Functional significance of leaf structure was examined by measures of CO(2) uptake and stomatal distributions. All species have Kranz anatomy and C(4)delta(13)C values. Freshwater species have thin leaves with small ridges on adaxial sides and stomata on both adaxial and abaxial sides. By contrast, salt marsh species have thick leaves with very pronounced ridges on the adaxial side and stomata located almost exclusively on adaxial leaf surfaces. Salt marsh species also have a thicker cuticle on the abaxial than on the adaxial side of leaves, and CO(2) uptake during photosynthesis is restricted to the adaxial leaf surface. Salt marsh species are adapted to controlling water loss by having stomata in leaf furrows on the adaxial side, which increases the boundary layer, and by having large leaf ridges that fit together as the leaf rolls during water stress. Differences in structural-functional features of photosynthesis in Spartina species are suggested to be related to adaptations to saline environments.