Improving drought-, salinity-, and heat-tolerance in transgenic plants by co-overexpressing Arabidopsis vacuolar pyrophosphatase gene AVP1 and Larrea Rubisco activase gene RCA.

The severity and frequency of many abiotic stresses such as drought, salinity and heat, cause substantial crop losses worldwide, which poses a serious challenge in food security. To increase crop production, new approaches are needed. Previous research has shown that overexpression of the tonoplast H+ pyrophosphatase gene AVP1 leads to improved drought and salt tolerance in transgenic plants. Other research showed that overexpression of thermotolerant ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase gene could maintain photosynthesis at higher temperatures, which contributes to higher heat tolerance in transgenic plants. In nature, abiotic stresses rarely come alone, instead these stresses often occur in various combinations. Therefore, it is desirable to make crops more tolerant to multiple stresses, which will likely lead to higher crop yield under various stress conditions. It is shown here that co-overexpression of the Arabidopsis gene AVP1 and the Larrea Rubisco activase gene RCA significantly increases drought, salinity and heat tolerance, resulting in higher biomass and seed yield than wild-type plants. AVP1/RCA co-overexpressing plants are as more drought- and salt-tolerant as AVP1-overexpressing plants, and as more heat-tolerant as RCA-overexpressing plants. More importantly, they produce higher seed yields than AVP1-overexpressing, RCA-overexpressing, and wild-type plants under combined drought and heat conditions.

Non-structural carbohydrate dynamics associated with antecedent stem water potential and air temperature in a dominant desert shrub.

Non-structural carbohydrates (NSCs) are necessary for plant growth and affected by plant water status, but the temporal dynamics of water stress impacts on NSC are not well understood. We evaluated how seasonal NSC concentrations varied with plant water status (predawn xylem water potential, Ψ) and air temperature (T) in the evergreen desert shrub Larrea tridentata. Aboveground sugar and starch concentrations were measured weekly or monthly for ~1.5 years on 6-12 shrubs simultaneously instrumented with automated stem psychrometers; leaf photosynthesis (Anet ) was measured monthly for 1 year. Leaf sugar increased during the dry, premonsoon period, associated with lower Ψ (greater water stress) and high T. Leaf sugar accumulation coincided with declines in leaf starch and stem sugar, suggesting the prioritization of leaf sugar during low photosynthetic uptake. Leaf starch was strongly correlated with Anet and peaked during the spring and monsoon seasons, while stem starch remained relatively constant except for depletion during the monsoon. Recent photosynthate appeared sufficient to support spring growth, while monsoon growth required the remobilization of stem starch reserves. The coordinated responses of different NSC fractions to water status, photosynthesis, and growth demands suggest that NSCs serve multiple functions under extreme environmental conditions, including severe drought.

Temporal shifts in iso/anisohydry revealed from daily observations of plant water potential in a dominant desert shrub.

Plant species are characterized along a spectrum of isohydry to anisohydry depending on their regulation of water potential (Ψ), but the plasticity of hydraulic strategies is largely unknown. The role of environmental drivers was evaluated in the hydraulic behavior of Larrea tridentata, a drought-tolerant desert shrub that withstands a wide range of environmental conditions. With a 1.5 yr time-series of 2324 in situ measurements of daily predawn and midday Ψ, the temporal variability of hydraulic behavior was explored in relation to soil water supply, atmospheric demand and temperature. Hydraulic behavior in Larrea was highly dynamic, ranging from partial isohydry to extreme anisohydry. Larrea exhibited extreme anisohydry under wet soil conditions corresponding to periods of high productivity, whereas partial isohydry was exhibited after prolonged dry or cold conditions, when productivity was low. Environmental conditions can strongly influence plant hydraulic behavior at relatively fast timescales, which enhances our understanding of plant drought responses. Although species may exhibit a dominant hydraulic behavior, variable environmental conditions can prompt plasticity in Ψ regulation, particularly for species in seasonally dry climates.

Antecedent soil water content and vapor pressure deficit interactively control water potential in Larrea tridentata.

Plant water potential Ψ is regulated by stomatal responses to atmospheric moisture demand D and soil water availability W, but the timescales of influence and interactions between these drivers of plant Ψ are poorly understood. Here, we quantify the effects of antecedent D and W on plant Ψ in the desert shrub Larrea tridentata. Repeated measurements of plant baseline water potential ΨB and diurnal water potential ΨD were analyzed in a Bayesian framework to evaluate the influence of antecedent D and W at daily and subdaily timescales. Both ΨB and ΨD exhibited negative, 2- to 4-d lagged responses to daily-scale D; conversely, plant ΨD responded almost instantaneously to subdaily D, though the direction of this response depended on antecedent moisture conditions. Plant ΨB and ΨD responded positively and immediately (no lag) to shallow W, which contrasts the negative, lagged (6-7 d) response to deep W. The changing sensitivity of ΨD to subdaily D highlights shifting modes of plant Ψ regulation: D effects on ΨD range from negative to neutral to positive depending on past conditions and time of day. Explicit consideration of antecedent conditions across multiple timescales can reveal important complexities in plant responses.

The role of vegetation-microclimate feedback in promoting shrub encroachment in the northern Chihuahuan desert.

Many arid and semi-arid landscapes around the world are affected by a shift from grassland to shrubland vegetation, presumably induced by climate warming, increasing atmospheric CO2 concentrations, and/or changing land use. This major change in vegetation cover is likely sustained by positive feedbacks with the physical environment. Recent research has focused on a feedback with microclimate, whereby cold intolerant shrubs increase the minimum nocturnal temperatures in their surroundings. Despite the rich literature on the impact of land cover change on local climate conditions, changes in microclimate resulting from shrub expansion into desert grasslands have remained poorly investigated. It is unclear to what extent such a feedback can affect the maximum extent of shrub expansion and the configuration of a stable encroachment front. Here, we focus on the case of the northern Chihuahuan desert, where creosotebush (Larrea tridentata) has been replacing grasslands over the past 100-150 years. We use a process-based coupled atmosphere-vegetation model to investigate the role of this feedback in sustaining shrub encroachment in the region. Simulations indicate that the feedback allows juvenile shrubs to establish in the grassland during average years and, once established, reduce their vulnerability to freeze-induced mortality by creating a warmer microclimate. Such a feedback is crucial in extreme cold winters as it may reduce shrub mortality. We identify the existence of a critical zone in the surroundings of the encroachment front, in which vegetation dynamics are bistable: in this zone, vegetation can be stable both as grassland and as shrubland. The existence of these alternative stable states explains why in most cases the shift from grass to shrub cover is found to be abrupt and often difficult to revert.

Seasonal stomatal behavior of a common desert shrub and the influence of plant neighbors.

Stomata simultaneously regulate plant carbon gain and water loss, and patterns of stomatal conductance (g(s)) provide insight into water use strategies. In arid systems, g(s) varies seasonally based on factors such as water availability and temperature. Moreover, the presence and species identity of neighboring plants likely affects g(s) of the focal plant by altering available soil water and microclimate conditions. We investigated stomatal behavior in Larrea tridentata, a drought-tolerant, evergreen shrub occurring throughout the arid southwestern United States. We measured g(s) in Larrea over multiple seasons in the presence of neighbors representing different woody species. The data were analyzed in the context of a commonly used phenomenological model that relates g(s) to vapor pressure deficit (D) to understand spatial and temporal differences in stomatal behavior. We found that g(s) in Larrea was affected by neighborhood association, and these effects varied seasonally. The greatest effect of neighborhood association on g(s) occurred during the winter period, where Larrea growing alone (without neighbors) had higher g(s) compared to Larrea growing with neighbors. Larrea's stomatal sensitivity to D and reference conductance (i.e., g(s) at D = 1 kPa) also differed significantly among different neighbor associations. Random effects indicated reference g(s) varied over short time scales (daily), while stomatal sensitivity showed little daily or seasonal variation, but was notably affected by neighbor associations such that neighboring species, especially trees, reduced Larrea's sensitivity to D. Overall, seasonal dynamics and neighborhood conditions appear critical to understanding temporal and spatial variation in Larrea's physiological behavior.

Freezing regime and trade-offs with water transport efficiency generate variation in xylem structure across diploid populations of Larrea sp. (Zygophyllaceae).

PREMISE OF THE STUDY: The impact of changing temperature regime on plant distributions may depend on the nature of physiological variation among populations. The arid-land genus Larrea spans habitats with a range of freezing frequency in North and South America. We hypothesized that variation in xylem anatomy among populations and species within this genus is driven by plasticity and trade-offs between safety from freeze-thaw embolism and water transport efficiency. METHODS: We measured vessel density and diameter distributions to predict freeze-thaw embolism and water transport capacity for high and low latitude populations of three Larrea species grown in the field and a greenhouse common garden. KEY RESULTS: Among field-grown L. divaricata, low latitude plants had larger mean vessel diameter and greater predicted freeze-thaw embolism, but higher water transport capacity compared with high latitude plants. Though high latitude L. tridentata and L. nitida had abundant smaller vessels, these plants also produced very large vessels and had semi ring-porous wood structure. Thus, their predicted embolism and water transport capacity were comparable to those of low latitude plants. Differences among field-grown and common-garden-grown plants demonstrate that plasticity contributes to population differentiation in xylem characters, though high latitude L. divaricata exhibited relatively lower plasticity. CONCLUSIONS: Our results indicate that a trade-off between transport safety and efficiency contributes substantially to variation in xylem structure within the genus Larrea. In addition, we suggest that xylem plasticity may play a role in negotiating these trade-offs, with implications for responses to future climate change.

Does a decade of elevated [CO2] affect a desert perennial plant community?

Understanding the effects of elevated [CO2 ] on plant community structure is crucial to predicting ecosystem responses to global change. Early predictions suggested that productivity in deserts would increase via enhanced water-use efficiency under elevated [CO2], but the response of intact arid plant communities to elevated [CO2 ] is largely unknown. We measured changes in perennial plant community characteristics (cover, species richness and diversity) after 10 yr of elevated [CO2] exposure in an intact Mojave Desert community at the Nevada Desert Free-Air CO2 Enrichment (FACE) Facility. Contrary to expectations, total cover, species richness, and diversity were not affected by elevated [CO2]. Over the course of the experiment, elevated [CO2] had no effect on changes in cover of the evergreen C3 shrub, Larrea tridentata; alleviated decreases in cover of the C4 bunchgrass, Pleuraphis rigida; and slightly reduced the cover of C3 drought-deciduous shrubs. Thus, we generally found no effect of elevated [CO2] on plant communities in this arid ecosystem. Extended drought, slow plant growth rates, and highly episodic germination and recruitment of new individuals explain the lack of strong perennial plant community shifts after a decade of elevated [CO2].

Variation in seedling freezing response is associated with climate in Larrea.

Variation in freezing severity is hypothesized to have influenced the distribution and evolution of the warm desert evergreen genus Larrea. If this hypothesis is correct, performance and survival of species and populations should vary predictably along gradients of freezing severity. If freezing environment changes in the future, the ability of Larrea to adapt will depend on the structure of variation for freezing resistance within populations. To test whether freezing responses vary among and within Larrea populations, we grew maternal families of seedlings from high and low latitude L. divaricata and high latitude L. tridentata populations in a common garden. We measured survival, projected plant area and dark-adapted chlorophyll fluorescence (F (v) /F (m)) before and after cold acclimation and for 2 weeks following a single freeze. We detected significant variation in freezing resistance among species and populations. Maternal family lines differed significantly in their responses to cold acclimation and/or freezing for two out of the three populations: among L. tridentata maternal families and among low latitude L. divaricata maternal families. There were no significant differences across maternal families of high latitude L. divaricata. Our results indicate that increased freezing resistance in high latitude populations likely facilitated historical population expansion of both species into colder climates, but this may have occurred to a greater extent for L. tridentata than for L. divaricata. Differences in the structure of variation for cold acclimation and freezing responses among populations suggest potential differences in their ability to evolve in response to future changes in freezing severity.

Maintenance of C sinks sustains enhanced C assimilation during long-term exposure to elevated [CO2] in Mojave Desert shrubs.

During the first few years of elevated atmospheric [CO(2)] treatment at the Nevada Desert FACE Facility, photosynthetic downregulation was observed in desert shrubs grown under elevated [CO(2)], especially under relatively wet environmental conditions. Nonetheless, those plants maintained increased A (sat) (photosynthetic performance at saturating light and treatment [CO(2)]) under wet conditions, but to a much lesser extent under dry conditions. To determine if plants continued to downregulate during long-term exposure to elevated [CO(2)], responses of photosynthesis to elevated [CO(2)] were examined in two dominant Mojave Desert shrubs, the evergreen Larrea tridentata and the drought-deciduous Ambrosia dumosa, during the eighth full growing season of elevated [CO(2)] treatment at the NDFF. A comprehensive suite of physiological processes were collected. Furthermore, we used C labeling of air to assess carbon allocation and partitioning as measures of C sink activity. Results show that elevated [CO(2)] enhanced photosynthetic performance and plant water status in Larrea, especially during periods of environmental stress, but not in Ambrosia. δ(13)C analyses indicate that Larrea under elevated [CO(2)] allocated a greater proportion of newly assimilated C to C sinks than Ambrosia. Maintenance by Larrea of C sinks during the dry season partially explained the reduced [CO(2)] effect on leaf carbohydrate content during summer, which in turn lessened carbohydrate build-up and feedback inhibition of photosynthesis. δ(13)C results also showed that in a year when plant growth reached the highest rates in 5 years, 4% (Larrea) and 7% (Ambrosia) of C in newly emerging organs were remobilized from C that was assimilated and stored for at least 2 years prior to the current study. Thus, after 8 years of continuous exposure to elevated [CO(2)], both desert perennials maintained their photosynthetic capacities under elevated [CO(2)]. We conclude that C storage, remobilization, and partitioning influence the responsiveness of these desert shrubs during long-term exposure to elevated [CO(2)].

Drought increases freezing tolerance of both leaves and xylem of Larrea tridentata.

Drought and freezing are both known to limit desert plant distributions, but the interaction of these stressors is poorly understood. Drought may increase freezing tolerance in leaves while decreasing it in the xylem, potentially creating a mismatch between water supply and demand. To test this hypothesis, we subjected Larrea tridentata juveniles grown in a greenhouse under well-watered or drought conditions to minimum temperatures ranging from -8 to -24 °C. We measured survival, leaf retention, gas exchange, cell death, freezing point depression and leaf-specific xylem hydraulic conductance (k1). Drought-exposed plants exhibited smaller decreases in gas exchange after exposure to -8 °C compared to well-watered plants. Drought also conferred a significant positive effect on leaf, xylem and whole-plant function following exposure to -15 °C; drought-exposed plants exhibited less cell death, greater leaf retention, higher k1 and higher rates of gas exchange than well-watered plants. Both drought-exposed and well-watered plants experienced 100% mortality following exposure to -24 °C. By documenting the combined effects of drought and freezing stress, our data provide insight into the mechanisms determining plant survival and performance following freezing and the potential for shifts in L. tridentata abundance and range in the face of changing temperature and precipitation regimes.

Shrub invasion decreases diversity and alters community stability in northern Chihuahuan Desert plant communities.

BACKGROUND: Global climate change is rapidly altering species range distributions and interactions within communities. As ranges expand, invading species change interactions in communities which may reduce stability, a mechanism known to affect biodiversity. In aridland ecosystems worldwide, the range of native shrubs is expanding as they invade and replace native grassland vegetation with significant consequences for biodiversity and ecosystem functioning. METHODOLOGY: We used two long-term data sets to determine the effects of shrub encroachment by Larrea tridentata on subdominant community composition and stability in formerly native perennial grassland dominated by Bouteloua eriopoda in New Mexico, USA. PRINCIPAL FINDINGS: Our results indicated that Larrea invasion decreased species richness during the last 100 years. We also found that over shorter temporal scales species-poor subdominant communities in areas invaded by Larrea were less stable (more variable in time) compared to species rich communities in grass-dominated vegetation. Compositional stability increased as cover of Bouteloua increased and decreased as cover of Larrea increased. SIGNIFICANCE: Changes in community stability due to altered interspecific interactions may be one mechanism by which biodiversity declines in grasslands following shrub invasion. As global warming increases, shrub encroachment into native grasslands worldwide will continue to alter species interactions and community stability both of which may lead to a decline in biodiversity.

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.