Carbon Isotope Composition of Nighttime Leaf-Respired CO2 in the Agricultural-Pastoral Zone of the Songnen Plain,Northeast China

Variations in the carbon isotope signature of leaf dark-respired CO₂ (δ¹³C_R) within a single night is a widely observed phenomenon. However, it is unclear whether there are plant functional type differences with regard to the amplitude of the nighttime variation in δ¹³C_R. These differences, if present, would be important for interpreting the short-term variations in the stable carbon signature of ecosystem respiration and the partitioning of carbon fluxes. To assess the plant functional type differences relating to the magnitude of the nighttime variation in δ¹³C_R and the respiratory apparent fractionation, we measured the δ¹³C_R, the leaf gas exchange, and the δ¹³C of the respiratory substrates of 22 species present in the agricultural-pastoral zone of the Songnen Plain, northeast China. The species studied were grouped into C₃ and C₄ plants, trees, grasses, and herbs. A significant nocturnal shift in δ¹³C_R was detected in 20 of the studied species, with the magnitude of the shift ranging from 1‰ to 5.8‰. The magnitude of the nighttime variation in δ¹³C_R was strongly correlated with the daytime cumulative carbon assimilation, which suggests that variation in δ¹³C_R were influenced, to some extent, by changes in the contribution of malate decarboxylation to total respiratory CO₂ flux. There were no differences in the magnitude of the nighttime variation in δ¹³C_R between the C₃ and C₄ plants, as well as among the woody plants, herbs and graminoids. Leaf respired CO₂ was enriched in ¹³C compared to biomass, soluble carbohydrates and lipids; however the magnitude of enrichment differed between 8 pm and 4 am, which were mainly caused by the changes in δ¹³C_R. We also detected the plant functional type differences in respiratory apparent fractionation relative to biomass at 4 am, which suggests that caution should be exercised when using the δ¹³C of bulk leaf material as a proxy for the δ¹³C of leaf-respired CO₂.     

deltaC Values in Maize Leaf Correlate with Phosphoenolpyruvate Carboxylase Levels

Values of δ¹³C and levels of phosphoenolpyruvate carboxylase and ribulose 1,5-bisphosphate carboxylase/oxygenase were analyzed in segments from the fourth leaf of young maize (Zea mays L.) plants. The δ¹³C values became significantly more negative from the base to the tip of the leaves. Phosphoenolpyruvate carboxylase levels and ribulose bisphosphate carboxylase levels both increased from the base to the tip. The principal effect of phosphoenolpyruvate carboxylase levels or δ¹³C should arise through its effect on the carboxylation/diffusion balance in the mesophyll. In this case, δ¹³C values should become more negative as phosphoenolpyruvate carboxylase levels increase, unless there are offsetting changes in stomatal aperture. The principal effect of ribulose bisphosphate carboxylase/oxygenase on δ¹³C should occur through its effect on the extent of leakage of CO₂ from the bundle sheath cells. In this case, δ¹³C values should become more positive as ribulose bisphosphate carboxylase levels increase. Accordingly, the variation in δ¹³C values seen in maize leaves appears to be the result of variations in the level of phosphoenolpyruvate carboxylase.     

Relationships between C3 Plant Foliar Carbon Isotope Composition and Element Contents of Grassland Species at High Altitudes on the Qinghai-Tibet Plateau,China

Relationships of foliar carbon isotope composition (δ¹³C) with foliar C, N, P, K, Ca, Mg contents and their ratios of 219 C₃ species leaf samples, obtained in August in 2004 to 2007 from 82 high altitude grassland sites on the Qinghai-Tibet Plateau China, were examined. This was done with reference to the proposition that foliar δ¹³C increases with altitude and separately for the life-form groups of graminoids, forbs and shrubs and for the genera Stipa and Kobresia. For all samples, foliar δ¹³C was negatively related to foliar K, P and ∑_{K+ Ca+ Mg}, and positively correlated to foliar C, C/N and C/P. The significance of these correlations differed for the taxonomic and life-form groups. Lack of a relationship of foliar δ¹³C with foliar N was inconsistent with the majority of studies that have shown foliar δ¹³C to be positively related to foliar N due to a decrease of C_i/C_a (the ratio between intercellular and atmospheric concentration of CO₂) and explained as a result of greater photosynthetic capacity at higher foliar N concentration. However this inconsistency relates to other high altitude studies that have found that photosynthetic capacity remains constant as foliar N increases. After accounting for the altitudinal relationship with foliar δ¹³C, of the elements only the K effect was significant and was most strongly expressed for Kobresia. It is concluded that factors critical to plant survival and growth at very high altitudes, such as low atmospheric pressure and low temperatures, may preclude expression of relationships between foliar δ¹³C and foliar elements that have been observed at lower altitudes.     

Effects of Ontogeny on δ13C of Plant- and Soil-Respired CO2 and on Respiratory Carbon Fractionation in C3 Herbaceous Species

Knowledge gaps regarding potential ontogeny and plant species identity effects on carbon isotope fractionation might lead to misinterpretations of carbon isotope composition (δ¹³C) of respired CO₂, a widely-used integrator of environmental conditions. In monospecific mesocosms grown under controlled conditions, the δ¹³C of C pools and fluxes and leaf ecophysiological parameters of seven herbaceous species belonging to three functional groups (crops, forage grasses and legumes) were investigated at three ontogenetic stages of their vegetative cycle (young foliage, maximum growth rate, early senescence). Ontogeny-related changes in δ¹³C of leaf- and soil-respired CO₂ and ¹³C/¹²C fractionation in respiration (Δ_R) were species-dependent and up to 7‰, a magnitude similar to that commonly measured in response to environmental factors. At plant and soil levels, changes in δ¹³C of respired CO₂ and Δ_R with ontogeny were related to changes in plant physiological status, likely through ontogeny-driven changes in the C sink to source strength ratio in the aboveground plant compartment. Our data further showed that lower Δ_R values (i.e. respired CO₂ relatively less depleted in ¹³C) were observed with decreasing net assimilation. Our findings highlight the importance of accounting for ontogenetic stage and plant community composition in ecological studies using stable carbon isotopes.     

Evidence of Physiological Decoupling from Grassland Ecosystem Drivers by an Encroaching Woody Shrub

Shrub encroachment of grasslands is a transformative ecological process by which native woody species increase in cover and frequency and replace the herbaceous community. Mechanisms of encroachment are typically assessed using temporal data or experimental manipulations, with few large spatial assessments of shrub physiology. In a mesic grassland in North America, we measured inter- and intra-annual variability in leaf δ¹³C in Cornus drummondii across a grassland landscape with varying fire frequency, presence of large grazers and topographic variability. This assessment of changes in individual shrub physiology is the largest spatial and temporal assessment recorded to date. Despite a doubling of annual rainfall (in 2008 versus 2011), leaf δ¹³C was statistically similar among and within years from 2008-11 (range of −28 to −27‰). A topography*grazing interaction was present, with higher leaf δ¹³C in locations that typically have more bare soil and higher sensible heat in the growing season (upland topographic positions and grazed grasslands). Leaf δ¹³C from slopes varied among grazing contrasts, with upland and slope leaf δ¹³C more similar in ungrazed locations, while slopes and lowlands were more similar in grazed locations. In 2011, canopy greenness (normalized difference vegetation index – NDVI) was assessed at the centroid of individual shrubs using high-resolution hyperspectral imagery. Canopy greenness was highest mid-summer, likely reflecting temporal periods when C assimilation rates were highest. Similar to patterns seen in leaf δ¹³C, NDVI was highest in locations that typically experience lowest sensible heat (lowlands and ungrazed). The ability of Cornus drummondii to decouple leaf physiological responses from climate variability and fire frequency is a likely contributor to the increase in cover and frequency of this shrub species in mesic grassland and may be generalizable to other grasslands undergoing woody encroachment.     

Fractionation of the stable isotopes of inorganic carbon by seagrasses

The δ¹³C values for seagrasses collected along the Texas Gulf Coast range from −10.9 to −11.4‰. These values are similar to the δ¹³C values of terrestrial C₄ plants, but seagrasses lack bundle sheath cells which are important in determining the δ¹³C values of C₄ plants. This work attempts to explain the reason the δ¹³C values of seagrasses resemble the δ¹³C values of C₄ plants.     

Influence of Ozone on the Stable Carbon Isotope Composition, deltaC, of Leaves and Grain of Spring Wheat (Triticum aestivum L.)

The relative composition of stable carbon isotopes, δ¹³C, was determined in flag leaves and grain of spring wheat (Triticum aestivum L. cv Albis) grown in open-top field fumigation chambers and exposed to different O₃ levels during the growing season. The aim of the study was to establish exposure-response relationships for the radiation-weighted seasonal mean O₃ concentration and δ¹³C (relative deviation of the ¹³C/¹²C ratio) values of the two plant parts. Samples were collected at harvest in 1986, 1987, and 1988. With increasing O₃ concentration, δ¹³C values increased (became less negative) proportionally. Year to year δ¹³C differences at equivalent O₃ concentrations were small. The shift in δ¹³C caused by O₃ was more pronounced in grain than in leaves. According to models of ¹³C discrimination in C₃ plants, these results indicate increasing limitation of photosynthesis by CO₂ diffusion relative to limitation by carboxylation with increasing O₃ exposure. This conclusion is not in agreement with results from gas exchange analysis. Water use efficiency in green flag leaves tended to decrease with increasing O₃, indicating a dominating effect of O₃ on CO₂ carboxylation.     

Photosynthetic carbon metabolism of a marine grass

The δ¹³C value of a tropical marine grass Thalassia testudinum is −9.04‰. This value is similar to the δ¹³C value of terrestrial tropical grasses. The δ¹³C values of the organic acid fraction, the amino acid fraction, the sugar fraction, malic acid, and glucose are: −11.2‰, −13.1‰, −10.1‰, −11.1‰, and −11.5‰, respectively. The δ¹³C values of malic acid and glucose of Thalassia are similar to the δ¹³C values of these intermediates in sorghum leaves and attest to the presence of the photosynthetic C₄-dicarboxylic acid pathway in this marine grass. The inorganic HCO₃⁻ for the growth of the grass fluctuates between −6.7 to −2.7‰ during the day. If CO₂ fixation in Thalassia is catalyzed by phosphoenolpyruvate carboxylase (which would result in a −3‰ fractionation between HCO₃⁻ and malic acid), the predicted δ¹³C value for Thalassia would be −9.7 to −5.7‰. This range is close to the observed range of −12.6 to −7.8‰ for Thalassia and agree with the operation of the C₄-dicarboxylic acid pathway in this plant. The early products of the fixation of HCO₃⁻ in the leaf sections are malic acid and aspartic acid which are similar to the early products of CO₂ fixation in C₄ terrestrial plants.     

Relative Roles of Soil Moisture,Nutrient Supply,Depth, and Mechanical Impedance in Determining Composition and Structure of Wisconsin Prairies

Ecologists have long classified Midwestern prairies based on compositional variation assumed to reflect local gradients in moisture availability. The best known classification is based on Curtis’ continuum index (CI), calculated using the presence of indicator species thought centered on different portions of an underlying moisture gradient. Direct evidence of the extent to which CI reflects differences in moisture availability has been lacking, however. Many factors that increase moisture availability (e.g., soil depth, silt content) also increase nutrient supply and decrease soil mechanical impedance; the ecological effects of the last have rarely been considered in any ecosystem. Decreased soil mechanical impedance should increase the availability of soil moisture and nutrients by reducing the root costs of retrieving both. Here we assess the relative importance of soil moisture, nutrient supply, and mechanical impedance in determining prairie composition and structure. We used leaf δ¹³C of C₃ plants as a measure of growing-season moisture availability, cation exchange capacity (CEC) x soil depth as a measure of mineral nutrient availability, and penetrometer data as a measure of soil mechanical impedance. Community composition and structure were assessed in 17 remnant prairies in Wisconsin which vary little in annual precipitation. Ordination and regression analyses showed that δ¹³C increased with CI toward “drier” sites, and decreased with soil depth and % silt content. Variation in δ¹³C among remnants was 2.0‰, comparable to that along continental gradients from ca. 500–1500 mm annual rainfall. As predicted, LAI and average leaf height increased significantly toward “wetter” sites. CI accounted for 54% of compositional variance but δ¹³C accounted for only 6.2%, despite the strong relationships of δ¹³C to CI and CI to composition. Compositional variation reflects soil fertility and mechanical impedance more than moisture availability. This study is the first to quantify the effects of soil mechanical impedance on community ecology.     

Gas Exchange, Stomatal Behavior, and deltaC Values of the flacca Tomato Mutant in Relation to Abscisic Acid

The relationship between stomatal conductance and capacity for assimilation was investigated in flacca, a mutant of tomato (Lycopersicon esculentum Mill.) that has abnormal stomatal behavior and low abscisic acid (ABA) content. The assimilation capacity, determined by measuring assimilation rate as a function of intercellular CO₂ pressure, did not differ in leaves of flacca and its parent variety, Rheinlands Ruhm (RR). On the other hand, stomatal conductance of flacca leaves was greater than that of RR, and could be phenotypically reverted by spraying with 30 micromolar ABA. Stomatal conductance of flacca leaves was also reduced by increasing CO₂ pressure, increasing leaf to air vapor pressure difference, and decreasing quantum flux, irrespective of ABA treatment.

The high conductance of flacca leaves resulted in a high intercellular CO₂ pressure. This allowed greater discrimination against ¹³CO₂, as evidenced by more negative δ ¹³C values for flacca as compared to RR. The δ ¹³C values of both flacca and RR plants as influenced by ABA treatment were consistent with predictions based on gas exchange measurements, using a recent model of discrimination.

     

Compensation point and isotopic characteristics of c(3)/c(4) intermediates and hybrids in panicum

Leaf CO₂ compensation points and stable hydrogen, oxygen and carbon isotope ratios were determined for Panicum species including C₃/C₄ intermediate photosynthesis plants, hybrids between C₃/C₄ intermediates and C₃ plants, C₃ and C₄ plants in the Panicum genus as well as several other C₃ and C₄ plants. C₃ plants had the highest compensation points, followed by hybrids, C₃/C₄ intermediates, and C₄ plants. δ¹³C values of cellulose nitrate and saponifiable lipids from C₄ plants were about 10‰ higher than those observed for cellulose nitrate and saponifiable lipids of C₃/C₄ intermediates, hybrids, and C₃ plants. Oxygen isotope ratios of cellulose as well as those of leaf water were similar for all plants. There was substantial variability in the δD values of cellulose nitrate among the plants studied. In contrast, such variability was not observed in δD values of water distilled from the leaves, nor in the δD values of the saponifiable lipids. Variability in δD values of cellulose nitrate from C₃/C₄ intermediates, hybrids, C₃, and C₄ plants is due to fractionations occurring during biochemical reactions specific to leaf carbohydrate metabolism.     

Climatic Control on Plant and Soil δ13C along an Altitudinal Transect of Lushan Mountain in Subtropical China: Characteristics and Interpretation of Soil Carbon Dynamics

Decreasing temperature and increasing precipitation along altitude gradients are typical mountain climate in subtropical China. In such a climate regime, identifying the patterns of the C stable isotope composition (δ¹³C) in plants and soils and their relations to the context of climate change is essential. In this study, the patterns of δ¹³C variation were investigated for tree leaves, litters, and soils in the natural secondary forests at four altitudes (219, 405, 780, and 1268 m a.s.l.) in Lushan Mountain, central subtropical China. For the dominant trees, both leaf and leaf-litter δ¹³C decreased as altitude increased from low to high altitude, whereas surface soil δ¹³C increased. The lower leaf δ¹³C at high altitudes was associated with the high moisture-related discrimination, while the high soil δ¹³C is attributed to the low temperature-induced decay. At each altitude, soil δ¹³C became enriched with soil depth. Soil δ¹³C increased with soil C concentrations and altitude, but decreased with soil depth. A negative relationship was also found between O-alkyl C and δ¹³C in litter and soil, whereas a positive relationship was observed between aromatic C and δ¹³C. Lower temperature and higher moisture at high altitudes are the predominant control factors of δ¹³C variation in plants and soils. These results help understand C dynamics in the context of global warming.     

Using the Stable Carbon and Nitrogen Isotope Compositions of Vervet Monkeys (Chlorocebus pygerythrus) to Examine Questions in Ethnoprimatology

This study seeks to understand how humans impact the dietary patterns of eight free-ranging vervet monkey (Chlorocebus pygerythrus) groups in South Africa using stable isotope analysis. Vervets are omnivores that exploit a wide range of habitats including those that have been anthropogenically-disturbed. As humans encroach upon nonhuman primate landscapes, human-nonhuman primate interconnections become increasingly common, which has led to the rise of the field of ethnoprimatology. To date, many ethnoprimatological studies have examined human-nonhuman primate associations largely in qualitative terms. By using stable carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope analysis, we use quantitative data to understand the degree to which humans impact vervet monkey dietary patterns. Based on initial behavioral observations we placed the eight groups into three categories of anthropogenic disturbance (low, mid, and high). Using δ¹³C and δ¹⁵N values we estimated the degree to which each group and each anthropogenically-disturbed category was consuming C₄ plants (primarily sugar cane, corn, or processed foods incorporating these crops). δ¹³C values were significantly different between groups and categories of anthropogenic-disturbance. δ¹⁵N values were significantly different at the group level. The two vervet groups with the highest consumption of C₄ plants inhabited small nature reserves, appeared to interact with humans only sporadically, and were initially placed in the mid level of anthropogenic-disturbance. However, further behavioral observations revealed that the high δ¹³C values exhibited by these groups were linked to previously unseen raiding of C₄ crops. By revealing these cryptic feeding patterns, this study illustrates the utility of stable isotopes analysis for some ethnoprimatological questions.     

Selective autophagic receptor NbNBR1 prevents NbRFP1-mediated UPS-dependent degradation of βC1 to promote geminivirus infection

Autophagy is an evolutionarily conserved, lysosomal/vacuolar degradation mechanism that targets cell organelles and macromolecules. Autophagy and autophagy-related genes have been studied for their antiviral and pro-viral roles in virus-infected plants. Here, we demonstrate the pro-viral role of a selective autophagic receptor NbNBR1 in geminivirus-infected Nicotiana benthamiana plants. The βC1 protein encoded by tomato yellow leaf curl China betasatellite (TYLCCNB) that is associated with tomato yellow leaf curl China virus (TYLCCNV) enhanced the expression level of NbNBR1. Then NbNBR1 interacted with βC1 to form cytoplasmic granules. Interaction of NbNBR1 with βC1 could prevent degradation of βC1 by the NbRFP1, an E3 ligase. Overexpression of NbNBR1 in N. benthamiana plants increased βC1 accumulation and promoted virus infection. In contrast, silencing or knocking out NbNBR1 expression in N. benthamiana suppressed βC1 accumulation and inhibited virus infection. A single amino acid substitution in βC1 (βC1^K4A) abolished its interaction with NbNBR1, leading to a reduced level of βC1^K4A. The TYLCCNV/TYLCCNB^K4A mutant virus caused milder disease symptoms and accumulated much less viral genomic DNAs in the infected plants. Collectively, the results presented here show how a viral satellite-encoded protein hijacks host autophagic receptor NbNBR1 to form cytoplasmic granules to protect itself from NbRFP1-mediated degradation and facilitate viral infection.     

Spatial patterns of leaf δ13C and δ15N of aquatic macrophytes in the arid zone of northwestern China

Analysis of stable isotope composition is an important tool in research on plant physiological ecology. However, large‐scale patterns of leaf‐stable isotopes for aquatic macrophytes have received considerably less attention. In this study, we examined the spatial pattern of stable isotopes of carbon (δ¹³C) and nitrogen (δ¹⁵N) of macrophytes leaves collected across the arid zone of northwestern China (approximately 2.4 × 10⁶ km²) and attempted to illustrate its relationship with environmental factors (i.e., temperature, precipitation, potential evapotranspiration, sediment total carbon and nitrogen). Our results showed that the mean values of the leaf δ¹³C and δ¹⁵N in the macrophytes sampled from the arid zone were −24.49‰ and 6.82‰, respectively, which were far less depleted than those measured of terrestrial plants. The order of averaged leaf δ¹³C from different life forms was as follows: submerged > floating‐leaved > emergent. Additionally, our studies indicated that the values of foliar δ¹³C values of all the aquatic macrophytes were only negatively associated with precipitation, but the foliar δ¹⁵N values were mainly associated with temperature, precipitation, and potential evapotranspiration. Therefore, we speculated that water‐relation factors are the leaf δ¹³C determinant of macrophytes in the arid zone of northwestern China, and the main factors affecting leaf δ¹⁵N values are the complex combination of water and energy factors.     

Shifts in the Carbon Metabolism of Xerosicyos danguyi H. Humb. (Cucurbitaceae) Brought About by Water Stress : I. General Characteristics

Xerosicyos danguyi H. Humb. (Cucurbitaceae) is an unusual leaf succulent endemic to Madagascar. Under well-watered conditions the plant exhibited Crassulacean acid metabolism (CAM), as characterized by large diurnal changes in titratable acidity, predominantly nighttime stomatal opening and CO₂ uptake, and high δ¹³C values. When plants were exposed to water stress for a minimum of a month, they shifted to a mode of carbon metabolism previously labeled CAM-idling. Under this mode of metabolism, the plants exhibited reduced stomatal opening, reduced CO₂ uptake, dampened diurnal fluctuations in titratable acidity, and no apparent changes in the δ¹³C values. Additionally, investigations showed that the stress hormones 1-aminocyclopropane-1-carboxylic acid (an ethylene precursor) and abscisic acid increased as much as 6-fold in the water-stressed plants. The results are discussed in relation to physiological significance and evolution of the CAM-idling mode of metabolism.     

Two categories of c/c ratios for higher plants

¹³C/¹²C ratios have been determined for plant tissue from 104 species representing 60 families. Higher plants fall into two categories, those with low δ_PDBI¹³C values (—24 to —34‰) and those with high δ ¹³C values (—6 to —19‰). Algae have δ ¹³C values of —12 to —23‰. Photosynthetic fractionation leading to such values is discussed.     

Carbon isotope ratios in ear parts of triticale : influence of grain filling

Four triticale (×Triticosecale Wittmack) genotypes were grown under rainfed conditions with limited irrigation support in Lleida in northeast Spain. For each variety, samples consisting of 10 tillers with half-sterilized spikes were taken three times from anthesis to maturity. Carbon isotope ratios (δ¹³C) were then determined in water extracts from ear bracts (glumes, paleas, and lemmas), awns and flag leaves, and in powdered kernels. For the half-sterilized spikes, carbon isotope analysis was carried out separately in bracts and awns from fertile and nonfertile spikelets. The δ¹³C in the water-soluble fraction of awns, glumes, and glumells from fruitless spikelets was significantly higher than that from fertile spikelets sampled at mid-grain filling. Differences in δ¹³C among sterile and fertile spikelets were not significant in samples taken a few days after anthesis or at maturity. These results are in accordance with some degree of refixation by awns and ear bracts of the CO₂ respired by grains during grain filling. There was progressively higher δ¹³C from flag leaf blades to awns, glumes, and glumells. This variation in δ¹³C along plant parts may be caused by differences in the ratio of assimilation rate to CO₂-diffusive conductance. Values of δ¹³C of mature kernels were between the values at anthesis and mid-grain filling for the water-soluble fraction of flag leaves and inner bracts and were fairly similar to those of glumes and awns.     

Stable isotope canopy effects for sympatric monkeys at Ta? Forest,C?te d'Ivoire

This study tests the hypothesis that vertical habitat preferences of different monkey species inhabiting closed canopy rainforest are reflected in oxygen isotopes. We sampled bone from seven sympatric cercopithecid species in the Taï forest, Côte d''Ivoire, where long-term study has established taxon-specific patterns of habitat use and diet. Modern rib samples (n = 34) were examined for oxygen (δ¹⁸O_ap) and carbon (δ¹³C_ap) from bone apatite (‘bioapatite’), and carbon (δ¹³C_co) and nitrogen (δ¹⁵N_co) from bone collagen. Results are consistent for C₃ feeders in a closed canopy habitat. Low irradiance and evapotranspiration, coupled with high relative humidity and recycled CO₂ in forest understory, contribute to observed isotopic variability. Both δ¹³C_co and δ¹³C_ap results reflect diet; however, δ¹³C values are not correlated with species preference for canopy height. By contrast, δ¹⁸O_ap results are correlated with mean observed height and show significant vertical partitioning between taxa feeding at ground, lower and upper canopy levels. This oxygen isotope canopy effect has important palaeobiological implications for reconstructing vertical partitioning among sympatric primates and other species in tropical forests.     

Hydraulic Failure Defines the Recovery and Point of Death in Water-Stressed Conifers

This study combines existing hydraulic principles with recently developed methods for probing leaf hydraulic function to determine whether xylem physiology can explain the dynamic response of gas exchange both during drought and in the recovery phase after rewatering. Four conifer species from wet and dry forests were exposed to a range of water stresses by withholding water and then rewatering to observe the recovery process. During both phases midday transpiration and leaf water potential (Ψ_leaf) were monitored. Stomatal responses to Ψ_leaf were established for each species and these relationships used to evaluate whether the recovery of gas exchange after drought was limited by postembolism hydraulic repair in leaves. Furthermore, the timing of gas-exchange recovery was used to determine the maximum survivable water stress for each species and this index compared with data for both leaf and stem vulnerability to water-stress-induced dysfunction measured for each species. Recovery of gas exchange after water stress took between 1 and >100 d and during this period all species showed strong 1:1 conformity to a combined hydraulic-stomatal limitation model (r² = 0.70 across all plants). Gas-exchange recovery time showed two distinct phases, a rapid overnight recovery in plants stressed to <50% loss of leaf hydraulic conductance (K_leaf) and a highly Ψ_leaf-dependent phase in plants stressed to >50% loss of K_leaf. Maximum recoverable water stress (Ψ_min) corresponded to a 95% loss of K_leaf. Thus, we conclude that xylem hydraulics represents a direct limit to the drought tolerance of these conifer species.