Reproductive compensatory photosynthesis in a semi-arid rangeland bunchgrass.

While increased foliar photosynthesis is well documented across many plant species in response to diverse modes of herbivory, the compensatory ability of photosynthetically active reproductive structures is unknown. To address this, we partially defoliated basal florets in seed heads of crested wheatgrass (Agropyron cristatum (L.) Gaertn.), an exotic Eurasian perennial bunchgrass widely distributed across North American sagebrush steppe. We followed direct and indirect responses by tracking post-clipping photosynthesis in clipped basal and unclipped distal florets, respectively, and comparing these to similar florets on unclipped seed heads. Compensatory photosynthesis was apparent 24 h after clipping; over the pre-anthesis period, clipped basal floret photosynthesis was + 62%, stomatal conductance was + 82%, and PSII photochemical yield was - 39% of unclipped controls. After anthesis, intact florets distal to clipped florets had modestly higher photosynthetic rates compared to controls, while basal floret rates did not differ between treatments. Compensatory photosynthesis reduced intrinsic water use efficiency (iWUE; photosynthesis/stomatal conductance) 68-40% below controls over pre- and post-anthesis periods, respectively. Specific mass (dry mass/area) of clipped florets was - 15% of controls, while florets distal to these had specific mass 11% greater than distal or basal florets on unclipped seed heads. These results suggest damaged basal florets provided carbon to unaffected distal florets. This could explain crested wheatgrass's ability to produce viable seeds under conditions limiting to native bunchgrasses, and presents a novel mechanism germane to the development of convergent drought- and grazing-tolerance traits important to arid and semi-arid rangeland plant community resilience to climate variability.

Using Genomic Selection to Develop Performance-Based Restoration Plant Materials.

Effective native plant materials are critical to restoring the structure and function of extensively modified ecosystems, such as the sagebrush steppe of North America's Intermountain West. The reestablishment of native bunchgrasses, e.g., bluebunch wheatgrass (Pseudoroegneria spicata [Pursh] À. Löve), is the first step for recovery from invasive species and frequent wildfire and towards greater ecosystem resiliency. Effective native plant material exhibits functional traits that confer ecological fitness, phenotypic plasticity that enables adaptation to the local environment, and genetic variation that facilitates rapid evolution to local conditions, i.e., local adaptation. Here we illustrate a multi-disciplinary approach based on genomic selection to develop plant materials that address environmental issues that constrain local populations in altered ecosystems. Based on DNA sequence, genomic selection allows rapid screening of large numbers of seedlings, even for traits expressed only in more mature plants. Plants are genotyped and phenotyped in a training population to develop a genome model for the desired phenotype. Populations with modified phenotypes can be used to identify plant syndromes and test basic hypotheses regarding relationships of traits to adaptation and to one another. The effectiveness of genomic selection in crop and livestock breeding suggests this approach has tremendous potential for improving restoration outcomes for species such as bluebunch wheatgrass.

Impacts of hydraulic redistribution on grass-tree competition vs facilitation in a semi-arid savanna.

A long-standing ambition in ecosystem science has been to understand the relationship between ecosystem community composition, structure and function. Differential water use and hydraulic redistribution have been proposed as one mechanism that might allow for the coexistence of overstory woody plants and understory grasses. Here, we investigated how patterns of hydraulic redistribution influence overstory and understory ecophysiological function and how patterns vary across timescales of an individual precipitation event to an entire growing season. To this end, we linked measures of sap flux within lateral and tap roots, leaf-level photosynthesis, ecosystem-level carbon exchange and soil carbon dioxide efflux with local meteorology data. The hydraulic redistribution regime was characterized predominantly by hydraulic descent relative to hydraulic lift. We found only a competitive interaction between the overstory and understory, regardless of temporal time scale. Overstory trees used nearly all water lifted by the taproot to meet their own transpirational needs. Our work suggests that alleviating water stress is not the reason we find grasses growing in the understory of woody plants; rather, other stresses, such as excessive light and temperature, are being ameliorated. As such, both the two-layer model and stress gradient hypothesis need to be refined to account for this coexistence in drylands.

Functional response of U.S. grasslands to the early 21st-century drought.

Grasslands across the United States play a key role in regional livelihood and national food security. Yet, it is still unclear how this important resource will respond to the prolonged warm droughts and more intense rainfall events predicted with climate change. The early 21st-century drought in the southwestern United States resulted in hydroclimatic conditions that are similar to those expected with future climate change. We investigated the impact of the early 21st-century drought on aboveground net primary production (ANPP) of six desert and plains grasslands dominated by C4 (warm season) grasses in terms of significant deviations between observed and expected ANPP. In desert grasslands, drought-induced grass mortality led to shifts in the functional response to annual total precipitation (P(T)), and in some cases, new species assemblages occurred that included invasive species. In contrast, the ANPP in plains grasslands exhibited a strong linear function of the current-year P(T) and the previous-year ANPP, despite prolonged warm drought. We used these results to disentangle the impacts of interannual total precipitation, intra-annual precipitation patterns, and grassland abundance on ANPP, and thus generalize the functional response of C4 grasslands to predicted climate change. This will allow managers to plan for predictable shifts in resources associated with climate change related to fire risk, loss of forage, and ecosystem services.

When vegetation change alters ecosystem water availability.

The combined effects of vegetation and climate change on biosphere-atmosphere water vapor (H2 O) and carbon dioxide (CO2 ) exchanges are expected to vary depending, in part, on how biotic activity is controlled by and alters water availability. This is particularly important when a change in ecosystem composition alters the fractional covers of bare soil, grass, and woody plants so as to influence the accessibility of shallower vs. deeper soil water pools. To study this, we compared 5 years of eddy covariance measurements of H2 O and CO2 fluxes over a riparian grassland, shrubland, and woodland. In comparison with the surrounding upland region, groundwater access at the riparian sites increased net carbon uptake (NEP) and evapotranspiration (ET), which were sustained over more of the year. Among the sites, the grassland used less of the stable groundwater resource, and increasing woody plant density decoupled NEP and ET from incident precipitation (P), resulting in greater exchange rates that were less variable year to year. Despite similar gross patterns, how groundwater accessibility affected NEP was more complex than ET. The grassland had higher respiration (Reco ) costs. Thus, while it had similar ET and gross carbon uptake (GEP) to the shrubland, grassland NEP was substantially less. Also, grassland carbon fluxes were more variable due to occasional flooding at the site, which both stimulated and inhibited NEP depending upon phenology. Woodland NEP was large, but surprisingly similar to the less mature, sparse shrubland, even while having much greater GEP. Woodland Reco was greater than the shrubland and responded strongly and positively to P, which resulted in a surprising negative NEP response to P. This is likely due to the large accumulation of carbon aboveground and in the surface soil. These long-term observations support the strong role that water accessibility can play when determining the consequences of ecosystem vegetation change.

Photochemical performance of reproductive structures in Great Basin bunchgrasses in response to soil-water availability.

Active restoration, especially seeding, is necessary in sagebrush steppe rangelands degraded by the spread and dominance of exotic invasive annual grasses, in part due to low, episodic seed production of native perennial bunchgrasses. In contrast, the widespread exotic bunchgrass, crested wheatgrass, readily produces viable seed cohorts. How soil-water availability affects the ecophysiology of the reproductive structures that may underlie these differences are unclear. To address this, we measured pre- and post-anthesis chlorophyll fluorescence parameters of optimal (F v/F m) and light-adapted PSII quantum yield (ϕ PSII) and ϕ PSII-derived electron transport rate (ETR) response to photosynthetic photon flux density (PPFD) in seed heads and flag leaves of watered and unwatered crested wheatgrass and squirreltail wild rye. Watering increased F v/F m in the sampled structures of both species, but ϕ PSII was similar between watering treatments. Pre- to post-anthesis F v/F m levels were maintained in crested wheatgrass seed heads but declined in flag leaves, with the opposite pattern apparent in squirreltail. Watering did not affect the ETR-PPFD response, but crested wheatgrass seed heads maintained higher ETR across saturating PPFD than did squirreltail. These findings suggest (i) photochemical efficiency is expressed in structures most closely associated with reproductive effort, and (ii) documented differences in seed head photosynthetic characteristics likely include some degree of allocation to individual floret photosynthetic capacity in addition to structural characteristics. We concluded that these physiological and structural differences may contribute to the differential ability of these species to establish from seed, and may help in effective plant material selection needed to improve restoration and conservation success in sagebrush steppe rangelands.

Inter- and under-canopy soil water, leaf-level and whole-plant gas exchange dynamics of a semi-arid perennial C4 grass.

It is not clear if tree canopies in savanna ecosystems exert positive or negative effects on soil moisture, and how these might affect understory plant carbon balance. To address this, we quantified rooting-zone volumetric soil moisture (θ(25 cm)), plant size, leaf-level and whole-plant gas exchange of the bunchgrass, bush muhly (Muhlenbergia porteri), growing under and between mesquite (Prosopis velutina) in a southwestern US savanna. Across two contrasting monsoon seasons, bare soil θ(25 cm) was 1.0-2.5% lower in understory than in the intercanopy, and was consistently higher than in soils under grasses, where θ(25 cm) was similar between locations. Understory plants had smaller canopy areas and volumes with larger basal diameters than intercanopy plants. During an above-average monsoon, intercanopy and understory plants had similar seasonal light-saturated leaf-level photosynthesis (A(net-sat)), stomatal conductance (g(s-sat)), and whole-plant aboveground respiration (R(auto)), but with higher whole-plant photosynthesis (GEP(plant)) and transpiration (T(plant)) in intercanopy plants. During a below-average monsoon, intercanopy plants had higher diurnally integrated GEP(plant), R(auto), and T(plant). These findings showed little evidence of strong, direct positive canopy effects to soil moisture and attendant plant performance. Rather, it seems understory conditions foster competitive dominance by drought-tolerant species, and that positive and negative canopy effects on soil moisture and community and ecosystem processes depends on a suite of interacting biotic and abiotic factors.

Growing season ecosystem and leaf-level gas exchange of an exotic and native semiarid bunchgrass.

The South African grass, Lehmann lovegrass (Eragrostis lehmanniana), may alter ecosystem processes across extensive semiarid grasslands and savannahs of western North America. We compared volumetric soil moisture (theta), total and green tissue leaf area index (LAI), ecosystem (i.e. whole-plant and soil), and leaf-level gas exchange of Lehmann lovegrass and the native bush muhly (Muhlenbergia porteri) over the 2008 monsoon season in a semiarid savanna in southern Arizona, USA, to see if these were consistent with high productivity associated with lovegrass invasive success. theta across 0-5 and 0-25 cm was higher while evapotranspiration (ET) was similar between lovegrass and bush muhly plots, except shortly after rainfall, when ET was 32-81% higher in lovegrass plots. Lehmann lovegrass had lower, quickly developing LAI with greater leaf proportions than bush muhly. When early season theta was high, net ecosystem CO(2) exchange (NEE) was similar, but as storm frequency and theta declined, NEE was more negative in lovegrass (-0.69 to -3.00 micromol m(-2) s(-1)) than bush muhly (+1.75 to -1.55 micromol m(-2) s(-1)). Ecosystem respiration (R (eco)) responded quickly to monsoon onset and late-season rains, and was lower in lovegrass (2.44-3.74 micromol m(-2) s(-1)) than bush muhly (3.60-5.3 micromol m(-2) s(-1)) across the season. Gross ecosystem photosynthesis (GEP) was greater in Lehmann lovegrass, concurrent with higher leaf-level photosynthesis and stomatal conductance. We conclude that canopy structure facilitates higher theta under Lehmann lovegrass, reducing phenological constraints and stomatal limitations to whole-plant carbon uptake through the short summer monsoon growing season.

Photosynthetic regulation in seed heads and flag leaves of sagebrush-steppe bunchgrasses.

Native sagebrush-steppe bunchgrass populations are threatened by the spread and dominance of exotic invasive annual grasses, in part due to low, episodic seed production. In contrast, the widespread exotic bunchgrass, crested wheatgrass, readily produces viable seed cohorts. The mechanisms underlying these differences are unclear. To address this, we measured seed head specific mass (g m-2) and net photosynthetic assimilation (A net) as a function of internal [CO2] (A/Ci curves) in pre- and post-anthesis seed heads and flag leaves of crested wheatgrass and four native bunchgrasses to determine if differences in allocation and photosynthetic characteristics of seed heads was consistent with differential reproductive success. Crested wheatgrass seed heads had 2-fold greater specific mass compared to the native grasses, concurrent with greater CO2-saturated photosynthesis (A max), mesophyll carboxylation efficiency (CE), and higher intrinsic water-use efficiency (WUE i ; A net/stomatal conductance (g s)), but with similar relative stomatal limitations to photosynthesis (RSL). Post-anthesis seed head A max, CE, RSL and g s decreased in native grasses, while crested wheatgrass RSL decreased and CE increased dramatically, likely due to tighter coordination between seed head structural changes with stomatal and biochemical dynamics. Our results suggest native sagebrush-steppe bunchgrasses have greater stomatal and structural constraints to reproductive photosynthesis, while the exotic grass has evolved seed heads functionally similar to leaves. This study shows elucidating reproduction-related ecophysiological mechanisms provide understanding of plant attributes that underlie restoration success and could help guide the development of native plant materials with functional attributes needed to overcome demographic bottlenecks that limit their restoration into degraded sagebrush-steppe.

Fate and effects of heavy metals in salt marsh sediments.

The fate and effects of selected heavy metals were examined in sediment from a restored salt marsh. Sediment cores densely covered with Spartina patens were collected and kept either un-amended or artificially amended with nickel (Ni) under standardized greenhouse conditions. Ni-amendment had no significant effect on the fate of other metals in sediments, however, it increased root uptake of the metals. Metal translocation into the shoots was small for all metals. Higher Ni concentrations in plants from amended cores were accompanied by seasonal reductions in plant biomass, photosynthetic capacity and transfer efficiency of open photosystem II reaction centers; these effects, however, were no longer significant at the end of the growing season. Root colonization by arbuscular mycorrhizal fungi (AMF) resembled that of natural salt marshes with up to 20% root length colonized. Although Ni-amendment increased AMF colonization, especially during vegetative growth, in general AMF were largely unaffected.

Dynamic response of plant chlorophyll fluorescence to light, water and nutrient availability.

Chlorophyll molecules absorb photosynthetic active radiation (PAR). The resulting excitation energy is dissipated by three competing pathways at the level of photosystem: (i) photochemistry (and, by extension, photosynthesis); (ii) regulated and constitutive thermal energy dissipation; and (iii) chlorophyll-a fluorescence (ChlF). Because the dynamics of photosynthesis modulate the regulated component of thermal energy dissipation (widely addressed as non-photochemical quenching (NPQ)), the relationship between photosynthesis, NPQ and ChlF changes with water, nutrient and light availability. In this study we characterised the relationship between photosynthesis, NPQ and ChlF when conducting light-response curves of photosynthesis in plants growing under different water, nutrient and ambient light conditions. Our goals were to test whether ChlF and photosynthesis correlate in response to water and nutrient deficiency, and determine the optimum PAR level at which the correlation is maximal. Concurrent gas exchange and ChlF light-response curves were measured for Camelina sativa (L.) Crantz and Triticum durum (L.) Desf plants grown under (i) intermediate light growth chamber conditions, and (ii) high light environment field conditions respectively. Plant stress was induced by withdrawing water in the chamber experiment, and applying different nitrogen levels in the field experiment. Our study demonstrated that ChlF was able to track the variations in photosynthetic capacity in both experiments, and that the light level at which plants were grown was optimum for detecting both water and nutrient deficiency with ChlF. The decrease in photosynthesis was found to modulate ChlF via different mechanisms depending on the treatment: through the action of NPQ in response to water stress, or through the action of changes in leaf chlorophyll concentration in response to nitrogen deficiency. This study provides support for the use of remotely sensed ChlF as a proxy to monitor plant stress dynamics from space.

The effects of parental CO2 environment on seed quality and subsequent seedling performance in Bromusrubens.

Seeds were collected and compared from parent plants of Bromusrubens L. (Poaceae), an exotic Mojave Desert annual grass, grown in ambient (360 µmol mol-1) and elevated (700 µmol mol-1) CO2 to determine if parental CO2 growth conditions affected seed quality. Performance of seeds developed on the above plants was evaluated to determine the influence of parental CO2 growth conditions on germination, growth rate, and leaf production. Seeds of B. rubens developed on parents grown in elevated CO2 had a larger pericarp surface area, higher C:N ratio, and less total mass than ambient-developed seeds. Parental CO2 environment did not have an effect on germination percentage or mean germination time, as determined by radicle emergence. Seedlings from elevated-CO2-developed seeds had a reduced relative growth rate and achieved smaller final mass over the same growth period. Elevated-CO2-developed seeds had smaller seed reserves than ambient seeds, as determined by growing seedlings in sterile media and monitoring senescence. It appears that increased seed C:N ratios associated with plants grown under elevated CO2 may have a major effect on seed quality (morphology, nutrition) and seedling performance (e.g., growth rate and leaf production). Since the invasive success of B. rubens is primarily due to its ability to rapidly germinate, increase leaf area and maintain a relatively high growth rate compared to native annuals and perennial grasses, reductions in seed quality and seedling performance in elevated CO2 may have significant impacts on future community composition in the Mojave Desert.

Ecophysiological consequences of contrasting microenvironments on the desiccation tolerant moss Tortula ruralis.

Tortula ruralis is a homoiochlorophyllous-desiccation-tolerant (HDT) moss that retains all pigments when dehydrated and rapidly recovers physiological function upon rehydration. This moss forms extensive cover in exposed and shaded areas in the sandy semi-arid grasslands of Central Europe. We hypothesized that contrasting drying regimes between these microhabitats would affect plant N status, constraints to gas exchange and growth, as well as result in altered pigment concentrations and ratios, and photochemical light-response dynamics. Furthermore, we believed T. ruralis's HDT habit would limit its ability to acclimate to altered light environment. We found that sun plant T. ruralis had lower plant mass, as well as lower tissue N, C, total photosynthetic pigment concentrations and carbon isotope discrimination (Δ) values compared to shade plant counterparts. Carotenoid/chlorophyll ratios in sun plants were typical of high light-adapted tissue, but chlorophyll a/chlorophyll b ratios were lower, more characteristic of low light-adapted tissue. This unique combination of pigment responses was accompanied by sustained lower levels of optimal quantum efficiency of PSII (F v/F m) in sun plant T. ruralis, even during favorable diurnal conditions, and reduced engagement of energy-dependent thermal dissipation (NPQ). Reciprocal transplants of sun and shade plants showed that T. ruralis is capable of short-term adjustment to altered light level, as evidenced by increases in F v/F m, NPQ, and light-adapted PSII yield (φPSII) in transplanted sun plants, and concurrent decreases in sun-transplanted shade plants. However, the performance of transplanted sun plants remained consistently below that of undisturbed shade plants. These findings show that microenvironmental variation results in different patterns of resource acquisition in this HDT moss, and that growth in the open imparts greater desiccation tolerance, and the development of a greater standing engagement of slowly reversing photoprotective mechanisms. In contrast, prolonged activity and greater resource acquisition in shaded populations may allow T. ruralis to rapidly adjust to changes following disturbance to the plant canopy, fostering the persistence of T. ruralis in these semi-arid grasslands.

Transgenic poplar characterized by ectopic expression of a pine cytosolic glutamine synthetase gene exhibits enhanced tolerance to water stress.

Physiological responses to water stress in hybrid poplar (INRA 7171-B4, Populus tremula L. x P. alba L.) lines transformed to overexpress a pine cytosolic glutamine synthetase (GS1) gene were compared with those of non-transgenic plants. Before, during and after a drought treatment, net photosynthetic rates (Anet) were higher in transgenic than in non-transgenic plants. Stomatal conductance (gs) was higher in transgenic than in non-transgenic plants before, but not after exposure to drought. Before drought treatment, a sudden reduction in photosynthetic photon flux caused a greater burst of CO2 efflux in transgenic than non-transgenic plants, indicating greater photorespiratory activity. Drought caused greater reductions in photochemical quenching, photosystem II (PSII) antennae transfer efficiency (Fv'/Fm') and light-adapted PSII yield (PhiPSII) in non-transgenic than in transgenic plants, especially at low irradiances. Antennae-based thermal dissipation was higher in transgenic plants than in non-transgenic plants both during the imposition of drought and 1 or 3 days after the relief of drought. Under severe water stress and subsequently, transgenic plants maintained a higher expression of glutamine synthetase, glutamate synthase and Rubisco and higher concentrations of chlorophyll and glycine than non-transgenic plants. These findings indicate that overexpression of pine cytosolic GS1 enhanced sustained photosynthetic electron transport capacity during severe stomatal limitation. The data also suggest that ectopic expression of cytosolic GS increases photorespiratory activity, and that this serves as a protective sink for electrons from photosynthetic reaction centers.

Interactions among plant species and microorganisms in salt marsh sediments.

The interactions among Spartina patens and sediment microbial populations and the interactions among Phragmites australis and sediment microbial populations were studied at monotypic sites in Piermont Marsh, a salt marsh of the Hudson River north of New York, N.Y., at key times during the growing season. Arbuscular mycorrhizal fungi (AMF) effectively colonized S. patens but not P. australis, and there were seasonal increases and decreases that coincided with plant growth and senescence (17 and 6% of the S. patens root length were colonized, respectively). In sediment samples from the Spartina site, the microbial community and specific bacterial populations were at least twice as large in terms of number and biomass as the microbial community and specific bacterial populations in sediment samples from the Phragmites site, and peak values occurred during reproduction. Members of the domain Bacteria, especially members of the alpha-, gamma-, and delta-subdivisions of the Proteobacteria, were the most abundant organisms at both sites throughout the growing season. The populations were generally more dynamic in samples from the Spartina site than in samples from the Phragmites site. No differences between the two sites and no differences during the growing season were observed when restriction fragment length polymorphism analyses of nifH amplicons were performed in an attempt to detect shifts in the diversity of nitrogen-fixing bacteria. Differences were observed only in the patterns generated by PCR or reverse transcription-PCR for samples from the Spartina site, suggesting that there were differences in the overall and active populations of nitrogen-fixing bacteria. Regression analyses indicated that there was a positive interaction between members of the delta-subdivision of the Proteobacteria and root biomass but not between members of the delta-subdivision of the Proteobacteria and macroorganic matter at both sites. In samples from the Spartina site, there were indications that there were bacterium-fungus interactions since populations of members of the alpha-subdivision of the Proteobacteria were negatively associated with AMF colonization and populations of members of the gamma-subdivision of the Proteobacteria were positively associated with AMF colonization.

Carbon isotope discrimination and foliar nutrient status of Larrea tridentata (creosote bush) in contrasting Mojave Desert soils.

We investigated the relationships between foliar stable carbon isotope discrimination (Delta), % foliar N, and predawn water potentials (psi(pd)) and midday stomatal conductance ( g(s)) of Larrea tridentata across five Mojave Desert soils with different age-specific surface and sub-surface horizon development and soil hydrologies. We wished to elucidate how this long-lived evergreen shrub optimizes leaf-level physiological performance across soils with physicochemical characteristics that affect the distribution of limiting water and nitrogen resources. We found that in young, coarse alluvial soils that permit water infiltration to deeper soil horizons, % foliar N was highest and Delta, g(s) and psi(pd) were lowest, while %N was lowest and Delta, g(s) and psi(pd) were highest in fine sandy soils; Larrea growing in older soils with well-developed surface and sub-surface horizons exhibited intermediate values for these parameters. Delta showed negative linear relationships with % N (R(2)=0.54) and a positive relationship with psi(pd) (R(2)=0.14). Multiple regression analyses showed a strong degree of multicolinearity of g(s) and Delta with psi(pd) and N, suggesting that soil-mediated distribution of co-limiting water and nitrogen resources was the primary determinant of stomatal behavior, which is the primary limitation to productivity in this shrub. These findings show that subtle changes in the soil medium plays a strong role in the spatial and temporal distribution and utilization of limiting water and nitrogen resources by this long-lived desert evergreen, and that this role can be detected through carbon isotope ratios.