Climate variability supersedes grazing to determine the anatomy and physiology of a dominant grassland species.

Grassland ecosystems are historically shaped by climate, fire, and grazing which are essential ecological drivers. These grassland drivers influence morphology and productivity of grasses via physiological processes, resulting in unique water and carbon-use strategies among species and populations. Leaf-level physiological responses in plants are constrained by the underlying anatomy, previously shown to reflect patterns of carbon assimilation and water-use in leaf tissues. However, the magnitude to which anatomy and physiology are impacted by grassland drivers remains unstudied. To address this knowledge gap, we sampled from three locations along a latitudinal gradient in the mesic grassland region of the central Great Plains, USA during the 2018 (drier) and 2019 (wetter) growing seasons. We measured annual biomass and forage quality at the plot level, while collecting physiological and anatomical traits at the leaf-level in cattle grazed and ungrazed locations at each site. Effects of ambient drought conditions superseded local grazing treatments and reduced carbon assimilation and total productivity in A. gerardii. Leaf-level anatomical traits, particularly those associated with water-use, varied within and across locations and between years. Specifically, xylem area increased when water was more available (2019), while xylem resistance to cavitation was observed to increase in the drier growing season (2018). Our results highlight the importance of multi-year studies in natural systems and how trait plasticity can serve as vital tool and offer insight to understanding future grassland responses from climate change as climate played a stronger role than grazing in shaping leaf physiology and anatomy.

Sterile Spikelets Contribute to Yield in Sorghum and Related Grasses.

Sorghum (Sorghum bicolor) and its relatives in the grass tribe Andropogoneae bear their flowers in pairs of spikelets in which one spikelet (seed-bearing or sessile spikelet [SS]) of the pair produces a seed and the other is sterile or male (staminate). This division of function does not occur in other major cereals such as wheat (Triticum aestivum) or rice (Oryza sativa). Additionally, one bract of the SS spikelet often produces a long extension, the awn, that is in the same position as, but independently derived from, that of wheat and rice. The function of the sterile spikelet is unknown and that of the awn has not been tested in Andropogoneae. We used radioactive and stable isotopes of carbon, RNA sequencing of metabolically important enzymes, and immunolocalization of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to show that the sterile spikelet assimilates carbon, which is translocated to the largely heterotrophic SS. The awn shows no evidence of photosynthesis. These results apply to distantly related species of Andropogoneae. Removal of sterile spikelets in sorghum significantly decreases seed weight (yield) by ∼9%. Thus, the sterile spikelet, but not the awn, affects yield in the cultivated species and fitness in the wild species.

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.

NMR-based Metabolomics to Study the Cold-acclimation Strategy of Two Miscanthus Genotypes.

INTRODUCTION: Abiotic stress is a major cause of yield loss in plant culture. Miscanthus, a perennial C4 grass, is now considered a major source of renewable energy, especially for biofuel production. During the first year of planting in Northern Europe, Miscanthus was exposed to frost temperature, which generated high mortality in young plants and large loss of yield. One strategy to avoid such loss is to apply cold-acclimation, which confers on plants a better resistance to low temperature. OBJECTIVES: The aim of this study is to describe the effect of a cold-acclimation period on the metabolome of two Miscanthus genotypes that vary in their frost sensitivity at the juvenile stage. Miscanthus × giganteus (GIG) is particularly sensitive to frost, whereas Miscanthus sinensis August Feder (AUG) is tolerant. MATERIALS AND METHODS: Polar metabolite extraction was performed on Miscanthus, grown in non-acclimated or cold-acclimated conditions. Extracts were analysed by 1 H-NMR followed by multivariate statistical analysis. Discriminant metabolites were identified. RESULTS: More than 40 metabolites were identified in the two Miscanthus genotypes. GIG and AUG showed a different metabolic background before cold treatment, probably related to their genetic background. After cold-acclimation, GIG and AUG metabolomes remained different. The tolerant genotype showed notably higher levels of accumulation in proline, sucrose and maltose when subjected to cold. CONCLUSION: These two genotypes seem to have a different adaptation strategy in cold conditions. The studied change in the metabolome concerns different types of molecules related to the cold-tolerant behaviour of Miscanthus. Copyright © 2016 John Wiley & Sons, Ltd.

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.

Impact of a short-term heat event on C and N relations in shoots vs. roots of the stress-tolerant C4 grass, Andropogon gerardii.

Global warming will increase heat waves, but effects of abrupt heat stress on shoot-root interactions have rarely been studied in heat-tolerant species, and abrupt heat-stress effects on root N uptake and shoot C flux to roots and soil remains uncertain. We investigated effects of a high-temperature event on shoot vs. root growth and function, including transfer of shoot C to roots and soil and uptake and translocation of soil N by roots in the warm-season drought-tolerant C4 prairie grass, Andropogon gerardii. We heated plants in the lab and field (lab=5.5days at daytime of 30+5 or 10°C; field=5days at ambient (up to 32°C daytime) vs. ambient +10°C). Heating had small or no effects on photosynthesis, stomatal conductance, leaf water potential, and shoot mass, but increased root mass and decreased root respiration and exudation per g. (13)C-labeling indicated that heating increased transfer of recently-fixed C from shoot to roots and soil (the latter likely via increased fine-root turnover). Heating decreased efficiency of N uptake by roots (uptake/g root), but did not affect total N uptake or the transfer of labeled soil (15)N to shoots. Though heating increased soil temperature in the lab, it did not do so in the field (10cm depth); yet results were similar for lab and field. Hence, acute heating affected roots more than shoots in this stress-tolerant species, increasing root mass and C loss to soil, but decreasing function per g root, and some of these effects were likely independent of direct effects from soil heating.

Bud production and dynamics of flowering and vegetative tillers in Andropogon gerardii (Poaceae): the role of developmental constraints.

PREMISE OF THE STUDY: Perennial grasses maintain aboveground tiller populations through vegetative reproduction via belowground buds and sexual reproduction via seed. The maintenance of a bud bank has important demographic consequences for perennial grasses. A tradeoff between these reproductive modes would be expected for a plant with limited resource availability. However, the ontogeny of the tiller could affect its ability to allocate between these two modes of reproduction. METHODS: Vegetative bud production and dynamics and tiller production were examined biweekly through an annual cycle on vegetative and flowering tillers of Andropogon gerardii. KEY RESULTS: Andropogon gerardii maintains a large reserve of dormant buds. Although vegetative and flowering tillers had similar bud phenology, flowering tillers produced larger numbers of buds of larger size, and transitioned a larger proportion of their buds to tiller, than did vegetative tillers. Therefore, a negative consequence of sexual reproduction on vegetative reproduction was not evident at the tiller level. A size threshold for floral induction likely exists that results in flowering tillers having more buds per tiller than vegetative tillers. The increased bud outgrowth of flowering tillers could be a result of their larger bud size or weaker apical dominance as compared to vegetative tillers. CONCLUSIONS: Plant development can place significant constraints on tradeoffs between the reproductive modes in perennial grasses and could affect their plasticity in plant reproductive allocation. Differences in developmental phenology and bud production between flowering and vegetative tillers may influence grass responses to environmental changes such as altered precipitation regimes or resource availability.

Land-use changes alter radiative energy and water vapor fluxes of a tall-grass Andropogon field and a savanna-woodland continuum in the Orinoco lowlands.

Changes in land use in the Orinoco lowlands affect the daily trends of energy and water vapor fluxes. We analyzed these fluxes along a disturbance gradient beginning from a cultivated tall-grass Andropogon field (S1) and extending over three savanna sites with increasing woody cover over herbaceous vegetation. The savanna sites encompass a herbaceous savanna (S2), a tree savanna (S3) and a woodland savanna (S4). In the wet season, there were differences in the radiation budget: seasonally averaged albedo for S1 (0.17) exceeded that of S2-S4 (0.13-0.14). Eddy covariance fluxes indicate that the partitioning of the daily net radiation (Rn) into sensible and latent heat (lambda E) fluxes depends on land use. During the wet season, evapotranspiration (i.e., lambda E) over the S1-S4 sites accounted for a variable fraction of Rn (i.e., 0.75, 0.52, 0.67 and 0.68, respectively). Therefore, the Bowen ratio was typically below 1. As the dry season progressed, the lambda E/Rn ratio decreased markedly with increasing air and canopy temperatures and air humidity mole fraction deficit. The maximum evaporation rate over the S1-S4 sites was 3.2, 2.5, 3.5 and 4.1 mm day(-1), respectively, and the annual values were 721, 538, 771 and 732 mm year(-1), respectively, equivalent to 49, 65, 52 and 88% of the rainfall. Soil water content fell from a maximum above 0.28 in the wet season to 0.030, 0.026, 0.030 and 0.028 m(3) m(-3) at sites S1-S4, respectively, in the dry season. Leaf area index was greatly reduced as herbaceous vegetation dried out.

Wild plants, Andropogon virginicus L. and Miscanthus sinensis Anders, are tolerant to multiple stresses including aluminum, heavy metals and oxidative stresses.

To isolate high-tolerant plants against aluminum (Al), heavy metals and/or oxidative stresses as a final goal, screening of Al tolerant plants from a collection of 49 wild plants was first of all performed in this study. Andropogon virginicus L. and Miscanthus sinensis Anders showed high Al tolerant phenotypes (more than 35% values in both relative root growth and germination frequency even under 900 microM Al concentration) in our screening. Al tolerance mechanisms in these two plants were characterized and the results suggested that (1) a transport system of toxic Al ions from root to shoot, (2) a suppression of Al accumulation in root tip region and (3) a suppression of oxidative damages by an induction of anti-peroxidation enzymes, such as superoxide dismutase (SOD) and catalase, were involved in the tolerance mechanisms. Six wild plants [Andropogon, Miscanthus, Dianthus japonicus Thunb, Echinochloa crus-galli (L.) Beauv, Reynoutria japonica Houtt, and Sporobolus fertilis (Steud.) W. Clayton] were furthermore tested for their sensitivity against heavy metal stresses and oxidative stresses. The two high Al tolerant plants, Andropogon and/or Miscanthus, showed tolerance to Cr, Zn, diamide or hydrogen peroxide, suggesting common tolerance mechanisms among the tested stresses. Reynoutria showed tolerance to diamide and hydrogen peroxide, Sporobolus to Cr and Echinocholoa to Cd and Cu. Moreover, the collection of wild plants used in this study was a very useful kit to isolate tolerant plants against various abiotic stresses within a short period of time.

Influence of root herbivory on plant communities in heterogeneous nutrient environments.

While plant species respond differently to nutrient patches, the forces that drive this variability have not been extensively examined. In particular, the role of herbivory in modifying plant-resource interactions has been largely overlooked. We conducted a glasshouse study in which nutrient heterogeneity and root herbivory were manipulated, and used differences in foraging among plant species to predict the influence of root herbivores on these species in competition. We also tracked the influence of neighborhood composition, heterogeneity, and herbivory on whole-pot plant biomass. When herbivores were added to mixed-species neighborhoods, Eupatorium compositifolium, the most precise forager, was the only plant species to display a reduction in shoot biomass. Neighborhood composition had the greatest influence on whole-pot biomass, followed by nutrient heterogeneity; root herbivory had the smallest influence. These results suggest that root herbivory is a potential cost of morphological foraging in roots. Root herbivores reduced standing biomass and influenced the relative growth of species in mixed communities, but their effect was not strong enough at the density examined to overwhelm the bottom-up effects of resource distribution.

Evaluation of microbial inoculation and vegetation to enhance the dissipation of atrazine and metolachlor in soil.

Four greenhouse studies were conducted to evaluate the effects of native prairie grasses and two pesticide-degrading bacteria to remediate atrazine and metolachlor in soils from agricultural dealerships (Alpha site soil, northwest Iowa, USA; Bravo site soil, central Iowa, USA). The Alpha soil contained a low population of atrazine-degrading microorganisms relative to the Bravo soil. Each soil freshly treated with atrazine or metolachlor was aged for a short or long period of time, respectively. An atrazine-degrading bacterium, Agrobacterium radiobacter strain J14a; a metolachlor-degrading bacterium, Pseudomonas fluorescens strain UA5-40; and a mixture of three native prairie grasses-big bluestem (Andropogon gerardii Vitman), yellow Indian grass (Sorghastrum nutans [L.] Nash), and switchgrass (Panicum virgatum L.)-were added to the soils after the soils were aged for long periods of time. The soils aged for short periods of time were treated with J14a, the prairie grasses, or both after aging. The J14a and the grasses significantly reduced the concentration of atrazine in Alpha soil when the soil was aged for a short period of time. However, these treatments had no statistically significant effect when the soil was aged for a long period of time or on atrazine in Bravo soil. Inoculation with UA5-40 did not enhance metolachlor dissipation in either soil, but vegetation did increase metolachlor dissipation. Our results indicate that the dissipation of atrazine by J14a is affected by the presence of indigenous atrazine-mineralizing microorganisms and probably by the bioavailability of atrazine in the soil.

Defoliation effects on the community structure of arbuscular mycorrhizal fungi based on 18S rDNA sequences.

The effects of defoliation on arbuscular mycorrhizal (AM) associations in the field were investigated in terms of the community structure of AM fungi colonizing roots of grassland plants; the carbohydrate balance of the host plants was also determined. We focused on two plant species dominating Japanese native grasslands: the grazing-intolerant species Miscanthus sinensis and the grazing-tolerant species Zoysia japonica. Community structures of AM fungi were determined from 18S rRNA gene sequences. The dominant fungal group in both plant species was the Glomus clade, which was classified into several phylogenetic groups based on genetic distances and topology. In Miscanthus roots, the Glomus-Ab, Glomus-Ac, and Glomus-Ad groups were detected almost equally. In Zoysia roots, the Glomus-Ab group was dominant. Defoliation effects on the community structure of AM fungi differed between the plant species. In Miscanthus roots, the percentage of root length colonized (%RLC) by the Glomus-Ac and Glomus-Ad groups was significantly reduced by defoliation treatment. On the other hand, AM fungal group composition in Zoysia roots was unaffected by defoliation except on the last sampling date. Decreased %RLC by Glomus-Ac and Glomus-Ad coincided with decreased non-structural carbohydrate (NSC) levels in host plants; also, significant positive correlations were found between the %RLC and some NSC levels. On the other hand, the %RLC by Glomus-Ab in both plant species was unaffected by the NSC level. These results suggest that AM fungal groups have different carbohydrate requirements from host plants.