Multidecadal records of intrinsic water-use efficiency in the desert shrub Encelia farinosa reveal strong responses to climate change.

While tree rings have enabled interannual examination of the influence of climate on trees, this is not possible for most shrubs. Here, we leverage a multidecadal record of annual foliar carbon isotope ratio collections coupled with 39 y of survey data from two populations of the drought-deciduous desert shrub Encelia farinosa to provide insight into water-use dynamics and climate. This carbon isotope record provides a unique opportunity to examine the response of desert shrubs to increasing temperature and water stress in a region where climate is changing rapidly. Population mean carbon isotope ratios fluctuated predictably in response to interannual variations in temperature, vapor pressure deficit, and precipitation, and responses were similar among individuals. We leveraged the well-established relationships between leaf carbon isotope ratios and the ratio of intracellular to ambient CO2 concentrations to calculate intrinsic water-use efficiency (iWUE) of the plants and to quantify plant responses to long-term environmental change. The population mean iWUE value increased by 53 to 58% over the study period, much more than the 20 to 30% increase that has been measured in forests [J. Peñuelas, J. G. Canadell, R. Ogaya, Glob. Ecol. Biogeogr. 20, 597-608 (2011)]. Changes were associated with both increased CO2 concentration and increased water stress. Individuals whose lifetimes spanned the entire study period exhibited increases in iWUE that were very similar to the population mean, suggesting that there was significant plasticity within individuals rather than selection at the population scale.

A tale of ENSO, PDO, and increasing aridity impacts on drought-deciduous shrubs in the Death Valley region.

Germination, establishment, phenology, and death among three drought-deciduous shrubs were influenced by ENSO/PDO and precipitation, based on 37 years of annual surveys. Encelia farinosa forms near monospecific stands on slopes, whereas E. frutescens and Ambrosia salsola dominate wash habitats. All shrubs exhibited phenological coherence. While germination, establishment, and mortality patterns were similar among wash species, these dynamics contrasted with E. farinosa on slopes. Germination was associated with El Niño years. Slope plant establishment was dependent on precipitation in the subsequent year, but not evidently so in wash species. Major mortality events were episodic, with Encelia mortality just as likely to occur in years with below or above average precipitation. In both Encelia species, mortality was associated with transitions to a cold PDO phase. In E. frutescens this response was more rapid, whereas in E. farinosa mortality lagged 1 year, resulting in contrasting slope-wash mortality patterns. 50% of newly established shrubs died within 5, 5, and 18 years for E. frutescens, E. farinosa, and A. salsola, respectively. The 90% mortality ages were 26 years for E. frutescens, 24 years for E. farinosa, and 51 years for A. salsola. While maximum life expectancies are unknown, estimated maximum life expectancies were 56, 66, and 86 years for E. frutescens, E. farinosa, and A. salsola, respectively. Overall, as the climate has become more arid over the past four decades, the populations in both slope and wash habitats have exhibited similar responses: reduced shrub abundances and reduced total supportable leaf areas.

Soil microbial responses to nitrogen addition in arid ecosystems.

The N cycle of arid ecosystems is influenced by low soil organic matter, high soil pH, and extremes in water potential and temperature that lead to open canopies and development of biological soil crusts (biocrusts). We investigated the effects of N amendment on soil microbial dynamics in a Larrea tridentata-Ambrosia dumosa shrubland site in southern Nevada USA. Sites were fertilized with a NO3-NH4 mix at 0, 7, and 15 kg N ha(-1) y(-1) from March 2012 to March 2013. In March 2013, biocrust (0-0.5 cm) and bulk soils (0-10 cm) were collected beneath Ambrosia canopies and in the interspaces between plants. Biomass responses were assessed as bacterial and fungal SSU rRNA gene copy number and chlorophyll a concentration. Metabolic responses were measured by five ecoenzyme activities and rates of N transformation. By most measures, nutrient availability, microbial biomass, and process rates were greater in soils beneath the shrub canopy compared to the interspace between plants, and greater in the surface biocrust horizon compared to the deeper 10 cm soil profile. Most measures responded positively to experimental N addition. Effect sizes were generally greater for bulk soil than biocrust. Results were incorporated into a meta-analysis of arid ecosystem responses to N amendment that included data from 14 other studies. Effect sizes were calculated for biomass and metabolic responses. Regressions of effect sizes, calculated for biomass, and metabolic responses, showed similar trends in relation to N application rate and N load (rate × duration). The critical points separating positive from negative treatment effects were 88 kg ha(-1) y(-1) and 159 kg ha(-1), respectively, for biomass, and 70 kg ha(-1) y(-1) and 114 kg ha(-1), respectively, for metabolism. These critical values are comparable to those for microbial biomass, decomposition rates and respiration reported in broader meta-analyses of N amendment effects in mesic ecosystems. However, large effect sizes at low N addition rates indicate that arid ecosystems are sensitive to modest increments in anthropogenic N deposition.

Glacial trees from the La Brea tar pits show physiological constraints of low CO2.

• While studies of modern plants indicate negative responses to low [CO2] that occurred during the last glacial period, studies with glacial plant material that incorporate evolutionary responses are rare. In this study, physiological responses to changing [CO2] were compared between glacial (La Brea tar pits) and modern Juniperus trees from southern California. • Carbon isotopes were measured on annual rings of glacial and modern Juniperus. The intercellular:atmospheric [CO2] ratio (c(i) /c(a) ) and intercellular [CO2] (c(i) ) were then calculated on an annual basis and compared through geologic time. • Juniperus showed constant mean c(i) /c(a) between the last glacial period and modern times, spanning 50,000 yr. Interannual variation in physiology was greatly dampened during the last glacial period relative to the present, indicating constraints of low [CO2] that reduced responses to other climatic factors. Furthermore, glacial Juniperus exhibited low c(i) that rarely occurs in modern trees, further suggesting limiting [CO2] in glacial plants. • This study provides some of the first direct evidence that glacial plants remained near their lower carbon limit until the beginning of the glacial-interglacial transition. Our results also suggest that environmental factors that dominate carbon-uptake physiology vary across geologic time, resulting in major alterations in physiological response patterns through time.

Past climate changes and ecophysiological responses recorded in the isotope ratios of saguaro cactus spines.

The stable isotope composition of spines produced serially from the apex of columnar cacti has the potential to be used as a record of changes in climate and physiology. To investigate this potential, we measured the delta(18)O, delta(13)C and F(14)C values of spines from a long-lived columnar cactus, saguaro (Carnegiea gigantea). To determine plant age, we collected spines at 11 different heights along one rib from the stem apex (3.77 m height) to the base of a naturally occurring saguaro. Fractions of modern carbon (F(14)C) ranged from 0.9679 to 1.5537, which is consistent with ages between 1950 and 2004. We observed a very strong positive correlation (r = 0.997) between the F(14)C age of spines and the age of spines determined from direct and repeated height measurements taken on this individual over the past 37 years. A series of 96 spines collected from this individual had delta(18)O values ranging from 38 per thousand to 50 per thousand [Vienna standard mean ocean water (VSMOW)] and delta(13)C values from -11.5 per thousand to -8.5 per thousand [Vienna Peedee belemnite (VPDB)]. The delta(18)O and delta(13)C values of spines were positively correlated (r = 0.45, P < 0.0001) and showed near-annual oscillations over the approximately 15-year record. This pattern suggests that seasonal periods of reduced evaporative demand or greater precipitation input may correspond to increased daytime CO(2) uptake. The lowest delta(18)O and delta(13)C values of spines observed occurred during the 1983 and 1993 El Niño years, suggesting that the stable isotope composition recorded in spine tissue may serve as a proxy for these climate events. We compared empirical models and data from potted experimental cacti to validate these observations and test our hypotheses. The isotopic records presented here are the first ever reported from a chronosequence of cactus spines and demonstrate that tissues of columnar cacti, and potentially other long-lived succulents, may contain a record of past physiological and climatic variation.

Functional diversity of carbon-gain, water-use, and leaf-allocation traits in trees of a threatened lowland dry forest in Hawaii.

We examined carbon-gain, water-use, and leaf-allocation traits for six tree species of a Hawaiian dry forest to better understand the functional diversity within this threatened ecosystem. Tropical dry forests are among the most endangered ecosystems on Earth, and in Hawaii, as elsewhere, declining biodiversity threatens ecosystem processes that may depend on forest functional diversity. We found broad variation among species including a two-fold difference for mean photosynthetic rate, a greater than three-fold difference for predawn water potential, and a nearly three-fold difference for leaf life span. Principal component analysis showed a clear separation of species based on carbon-gain vs. water-use related axes, and δ(13)C analysis revealed differing limitations (supply vs. demand) on carbon assimilation. The broad functional variation not only spanned traditional classifications (avoiders vs. tolerators), but also included unusual strategies (e.g., fast growth with drought tolerance). Correlations among traits, including leaf life span, leaf mass per area, and %N, followed typical global patterns, but some exceptions appeared as a result of unique life-history characteristics, such as latex-rich sap and root parasitism. Elucidating functional variation provides important information that can be used to link plant biodiversity with ecosystem processes and also facilitate the management and preservation of tropical dry forests and other threatened communities.

Comparative analysis of cellulose preparation techniques for use with 13C, 14C, AND 18O isotopic measurements.

A number of operationally defined methods exist for pretreating plant tissues in order to measure C, N, and O isotopes. Because these isotope measurements are used to infer information about environmental conditions that existed at the time of tissue growth, it is important that these pretreatments remove compounds that may have exchanged isotopes or have been synthesized after the original formation of these tissues. In stable isotope studies, many pretreatment methods focus on isolating "cellulose" from the bulk tissue sample because cellulose does not exchange C and O isotopes after original synthesis. We investigated the efficacy of three commonly applied pretreatment methods, the Brendel method and two variants of the Brendel method, the Jayme-Wise method and successive acid/base/acid washes, for use on three tissue types (wood, leaves, roots). We then compared the effect of each method on C and O isotope composition (13C, 14C, 18O), C and N content, and chemical composition of the residue produced (using 13C nuclear magnetic resonance (NMR)). Our results raised concerns over use of the Brendel method as published, as it both added C and N to the sample and left a residue that contains remnant lipids and waxes. Furthermore, this method resulted in 18O values that are enriched relative to the other methods. Modifying the Brendel method by adding a NaOH step (wash) solved many of these problems. We also found that processed residues vary by tissue type. For wood and root tissues, the 13C NMR spectra and the 18O and 13C data showed only small differences between residues for the Jayme-Wise and modified Brendel methods. However, for leaf tissue, 13C NMR data showed that Jayme-Wise pretreatments produced residues that are more chemically similar to cellulose than the other methods. The acid/base/acid washing method generated 13C NMR spectra with incomplete removal of lignin for all tissues tested and both isotopic, and 13C NMR results confirmed that this method should not be used if purified cellulose is desired.

Precipitation pulses and carbon fluxes in semiarid and arid ecosystems.

In the arid and semiarid regions of North America, discrete precipitation pulses are important triggers for biological activity. The timing and magnitude of these pulses may differentially affect the activity of plants and microbes, combining to influence the C balance of desert ecosystems. Here, we evaluate how a "pulse" of water influences physiological activity in plants, soils and ecosystems, and how characteristics, such as precipitation pulse size and frequency are important controllers of biological and physical processes in arid land ecosystems. We show that pulse size regulates C balance by determining the temporal duration of activity for different components of the biota. Microbial respiration responds to very small events, but the relationship between pulse size and duration of activity likely saturates at moderate event sizes. Photosynthetic activity of vascular plants generally increases following relatively larger pulses or a series of small pulses. In this case, the duration of physiological activity is an increasing function of pulse size up to events that are infrequent in these hydroclimatological regions. This differential responsiveness of photosynthesis and respiration results in arid ecosystems acting as immediate C sources to the atmosphere following rainfall, with subsequent periods of C accumulation should pulse size be sufficient to initiate vascular plant activity. Using the average pulse size distributions in the North American deserts, a simple modeling exercise shows that net ecosystem exchange of CO2 is sensitive to changes in the event size distribution representative of wet and dry years. An important regulator of the pulse response is initial soil and canopy conditions and the physical structuring of bare soil and beneath canopy patches on the landscape. Initial condition influences responses to pulses of varying magnitude, while bare soil/beneath canopy patches interact to introduce nonlinearity in the relationship between pulse size and soil water response. Building on this conceptual framework and developing a greater understanding of the complexities of these eco-hydrologic systems may enhance our ability to describe the ecology of desert ecosystems and their sensitivity to global change.

Carbon isotope discrimination differences within and between contrasting populations of Encelia farinosa raised under common-environment conditions.

Previous studies of the desert shrub Encelia farinosa have shown variation of morpholological and physiological integration that appears to match environmental differences among populations. Such findings led us to ask if there is a genetic basis for such differentiation that may be related to physiological control of intercellular CO(2) concentrations as indicated by carbon isotope discrimination (Delta) values, and if genetic variance for Delta is detectable within populations. Under common environment conditions, Delta values were compared between two populations of E. farinosa from desert regions with contrasting rainfall patterns: Superior, Ariz., a region with high annual rainfall and droughts of short duration, and Oatman, Ariz. a region with lower annual rainfall and longer drought periods. Superior plants had consistently greater mean Delta values than Oatman plants across a broad range of soil water potentials, indicating that there is a genetic basis for Delta variation between these populations. At the intrapopulation level only Oatman plants showed detectable genetic variance of Delta based on: (1) consistent individual-rank values for Delta among soil-drought stages, and (2) evidence of heritable genetic variance for Delta during one drought stage. No genetic variance in Delta was evident for the Superior population. It is hypothesized that the high spatio-temporal heterogeneity of water availability at Oatman may facilitate the maintenance of genetic variance for carbon isotope discrimination within this population. Both the inter- and intra-population level findings suggest that selection associated with rainfall and drought has resulted in genetic divergence of the physiological factors involved in Delta determination for these populations. There appears to be strong differences of water-use and carbon-gain strategies among populations, and broader functional breadth among plants in the habitat of greatest environmental heterogeneity.

Population- and family-level variation of brittlebush (Encelia farinosa, Asteraceae) pubescence: its relation to drought and implications for selection in variable environments.

Because leaf pubescence of the desert shrub Encelia farinosa increases in response to drought and influences photosynthesis and transpiration, we hypothesized that differences in water availability across the range of this species may result in genetic differentiation for pubescence and associated productivity traits. We examined maternal family variation of pubescence-moderated light absorption (absorptance) in three populations of E. farinosa. Absorptance was always greatest for plants from the high-rainfall environment and lowest for those from the driest site, but the rate of absorptance change in response to drought was similar among all populations. Similar patterns were found when we compared families within populations-all genotypes had similar initial leaf absorptances, differentiated very early in seasonal growth, then had concordant changes in absorptance thereafter. However, family-level variance was greatest for plants from the driest site, a region with highly heterogeneous precipitation patterns, whereas low variance was found for plants from the wettest, least heterogeneous site. The concordance of leaf absorptance changes, within and among populations, may be due to integration with other drought-related traits; however, the differences in absorptance values within and among populations suggest that variation of leaf pubescence results from selection associated with geographical and local patterns of water availability.

Heavy and light beer: a carbon isotope approach to detect C(4) carbon in beers of different origins, styles, and prices.

The carbon isotope ratios (delta(13)C) of 160 beers from around the world ranged from -27.3 to -14.9 per thousand, primarily due to variation in the percentage of C(3) or C(4) plant carbon in the final product. Thirty-one percent of beers had a carbon signature of C(3) plants (barley, rice, etc.), whereas the remaining 69% contained some C(3)-C(4) mixture (mean of mixtures, 39 +/- 11% C(4) carbon). Use of C(4) carbon (corn, cane sugar, etc.) was not confined to beers from any particular region (Pacific Rim, Mexico, Brazil, Europe, Canada, and the United States). However, the delta(13)C of European beers indicated mostly C(3) plant carbon. In contrast, U.S. and Canadian beers contained either only C(3) or C(3)-C(4) mixtures; Brazilian, Mexican, and Pacific Rim beers were mostly C(3)-C(4) mixtures. Among different lagers, U.S.-style lagers generally contained more C(4) carbon than did imported pilsners. Among different ales, those brewed by large high-production breweries contained significant proportions of C(4) carbon, while C(4) carbon was not detected in microbrewery or home-brew ales. Furthermore, inexpensive beers generally contained more C(4) carbon than expensive beers.

Intraspecific variation of drought adaptation in brittlebush: leaf pubescence and timing of leaf loss vary with rainfall.

Reflective leaf pubescence of the desert shrub Encelia farinosa (brittlebrush) reduces leaf temperature and plant water loss, and is considered adaptive in xeric environments. Yet, little is known about intraspecific variation in this trait. Among three populations in the northern range of E. farinosa, which span a very broad precipitation gradient, both leaf absorptance variation and differences in the timing of drought-induced leaf loss were broadly associated with climatic variability. Where mean annual rainfall was greatest, drought-induced leaf loss was earliest, but these plants also had higher population-level mean leaf absorptance values. Higher absorptance increases the relative dependence on latent heat transfer (transpirational cooling), but it also provides greater instantaneous carbon assimilation. Plants at the driest site reached lower leaf absorptance values and maintained leaves longer into the drought period. Lower leaf absorptance reduces water consumption, and extended leaf longevity may buffer against the unpredictability of growing conditions experienced in the driest site. These observations are consistent with a trade-off scenario in which plants from wetter regions might trade off water conservation for higher instantaneous carbon gain, whereas plants from drier regions reduce water consumption and extend leaf longevity to maintain photosynthetic activity in the face of unpredictable growing conditions.

Comparisons of carbon isotope discrimination in populations of aridland plant species differing in lifespan.

Carbon isotope discrimination (Δ) was compared between populations of dominant perennial plant species, differing in life expectancy, in two deserts with contrasting vegetation types. In both deserts, plants of the shorter-lived species showed significantly higher Δ and greater intrapopulation variance in this character compared to the long-lived species. These results indicate underlying differences in gas-exchange physiology, and suggest a positive correlation between water-use efficiency and lifespan in desert plants. Differences in variance for this character may reflect greater microenvironmental variation experienced by shorter-lived plants and/or different forms of selection acting on water-use traits. Spatial distributions were significantly clustered for the shorter-lived species and significantly uniform for the long-lived species, indicating that competition has been important in the development of the long-lived populations. The long-lived Larrea tridentata showed a significant, negative correlation between Δ and Thiessen polygon area, suggesting a positive relationship between water-use efficiency and longevity within this species. This relationship was weakly supported in the other warm desert species, Encelia farinosa, but was not observed within populations of the cold desert species, Gutierrezia microcephala and Coleogyne ramosissima. These results suggest that Δ reflects key aspects of plant metabolism related to lifespan; these differences may ultimately influence interactions among desert plants and the structure of desert plant communities.

Differential utilization of summer rains by desert plants.

Seasonal changes in the hydrogen isotope ratios of xylem waters were measured to determine water sources used for growth in desert plants of southern Utah. While all species used winter-spring recharge precipitation for spring growth, utilization of summer rains was life-form dependent. Annuals and succulent perennials exhibited a complete dependence on summer precipitation. Herbaceous and woody perennial species simultaneously utilized both summer precipitation and remaining winter-spring precipitation, with herbaceous species much more reliant on the summer precipitation component. Several of the woody perennials exhibited no response to summer precipitation. Currently, precipitation in southern Utah is evenly partitioned between winter and summer time periods; however, global circulation models predict that summer precipitation will increase in response to anticipated climate change. Our data indicate that components within the community will differentially responde to the change in precipitation patterns. These results are discussed in relation to impact on competition and possible changes in community structure.