Renee A. Smith

Impacts of drought on leaf respiration in darkness and light in Eucalyptus saligna exposed to industrial‐age atmospheric CO2 and growth temperature

Our study assessed the impact of a wide range of industrial-age climate scenarios on leaf respiration (R) in Eucalyptus saligna. Well-watered or sustained drought-treated plants were grown in glasshouses differing in atmospheric CO₂ concentration ([CO₂]) (280, 400 and 640 μl l⁻¹) and temperature (26 and 30°C). Rates of R in darkness (R(dark) ) and light (R(light) ), photosynthesis (A) and related leaf traits (mass : area relationships, and nitrogen, phosphorus, starch and sugar concentrations) were measured. Light inhibited R in all cases (R(light) < R(dark) ) (well-watered: 40%; drought-treated: 73%). Growth [CO₂] and temperature had little impact on area-based rates of R(dark) or R(light) , with R(light) exhibiting minimal thermal acclimation. By contrast, sustained drought resulted in reduced R(dark), R(light) and A, with the inhibitory effect of drought on A and R(light) (c. 50-70%) greater than that on R(dark) (c. 15%). Drought effects were fully reversible after watering. Variability in R(light) appeared to be dependent on the underlying rate of R(dark) and associated Rubisco activity. Collectively, our data suggest that there is an asynchronous response of leaf carbon metabolism to drought, and a tighter coupling between R(light) and A than between R(dark) and A, under both past and future climate scenarios. These findings have important implications for ecosystem/global models seeking to predict carbon cycling.

Nocturnal stomatal conductance responses to rising [CO2], temperature and drought

The response of nocturnal stomatal conductance (g(s,n)) to rising atmospheric CO(2) concentration ([CO(2)]) is currently unknown, and may differ from responses of daytime stomatal conductance (g(s,d)). Because night-time water fluxes can have a significant impact on landscape water budgets, an understanding of the effects of [CO(2)] and temperature on g(s,n) is crucial for predicting water fluxes under future climates. Here, we examined the effects of [CO(2)] (280, 400 and 640 μmol mol(-1)), temperature (ambient and ambient + 4°C) and drought on g(s,n,) and g(s,d) in Eucalyptus sideroxylon saplings. g(s,n) was substantially higher than zero, averaging 34% of g(s,d). Before the onset of drought, g(s,n) increased by 85% when [CO(2)] increased from 280 to 640 μmol mol(-1), averaged across both temperature treatments. g(s,n) declined with drought, but an increase in [CO(2)] slowed this decline. Consequently, the soil water potential at which g(s,n) was zero (Ψ(0)) was significantly more negative in elevated [CO(2)] and temperature treatments. g(s,d) showed inconsistent responses to [CO(2)] and temperature. g (s,n) may be higher in future climates, potentially increasing nocturnal water loss and susceptibility to drought, but cannot be predicted easily from g(s,d). Therefore, predictive models using stomatal conductance must account for both g(s,n) and g(s,d) when estimating ecosystem water fluxes.

Non-structural carbohydrates in woody plants compared among laboratories

Non-structural carbohydrates (NSC) in plant tissue are frequently quantified to make inferences about plant responses to environmental conditions. Laboratories publishing estimates of NSC of woody plants use many different methods to evaluate NSC. We asked whether NSC estimates in the recent literature could be quantitatively compared among studies. We also asked whether any differences among laboratories were related to the extraction and quantification methods used to determine starch and sugar concentrations. These questions were addressed by sending sub-samples collected from five woody plant tissues, which varied in NSC content and chemical composition, to 29 laboratories. Each laboratory analyzed the samples with their laboratory-specific protocols, based on recent publications, to determine concentrations of soluble sugars, starch and their sum, total NSC. Laboratory estimates differed substantially for all samples. For example, estimates for Eucalyptus globulus leaves (EGL) varied from 23 to 116 (mean = 56) mg g(-1) for soluble sugars, 6-533 (mean = 94) mg g(-1) for starch and 53-649 (mean = 153) mg g(-1) for total NSC. Mixed model analysis of variance showed that much of the variability among laboratories was unrelated to the categories we used for extraction and quantification methods (method category R(2) = 0.05-0.12 for soluble sugars, 0.10-0.33 for starch and 0.01-0.09 for total NSC). For EGL, the difference between the highest and lowest least squares means for categories in the mixed model analysis was 33 mg g(-1) for total NSC, compared with the range of laboratory estimates of 596 mg g(-1). Laboratories were reasonably consistent in their ranks of estimates among tissues for starch (r = 0.41-0.91), but less so for total NSC (r = 0.45-0.84) and soluble sugars (r = 0.11-0.83). Our results show that NSC estimates for woody plant tissues cannot be compared among laboratories. The relative changes in NSC between treatments measured within a laboratory may be comparable within and between laboratories, especially for starch. To obtain comparable NSC estimates, we suggest that users can either adopt the reference method given in this publication, or report estimates for a portion of samples using the reference method, and report estimates for a standard reference material. Researchers interested in NSC estimates should work to identify and adopt standard methods.

Elevated [CO2] does not ameliorate the negative effects of elevated temperature on drought-induced mortality in Eucalyptus radiata seedlings.

It has been reported that elevated temperature accelerates the time-to-mortality in plants exposed to prolonged drought, while elevated [CO(2)] acts as a mitigating factor because it can reduce stomatal conductance and thereby reduce water loss. We examined the interactive effects of elevated [CO(2)] and temperature on the inter-dependent carbon and hydraulic characteristics associated with drought-induced mortality in Eucalyptus radiata seedlings grown in two [CO(2)] (400 and 640 μL L(-1)) and two temperature (ambient and ambient +4 °C) treatments. Seedlings were exposed to two controlled drying and rewatering cycles, and then water was withheld until plants died. The extent of xylem cavitation was assessed as loss of stem hydraulic conductivity. Elevated temperature triggered more rapid mortality than ambient temperature through hydraulic failure, and was associated with larger water use, increased drought sensitivities of gas exchange traits and earlier occurrence of xylem cavitation. Elevated [CO(2)] had a negligible effect on seedling response to drought, and did not ameliorate the negative effects of elevated temperature on drought. Our findings suggest that elevated temperature and consequent higher vapour pressure deficit, but not elevated [CO(2)], may be the primary contributors to drought-induced seedling mortality under future climates.

Climate determines vascular traits in the ecologically diverse genus Eucalyptus

Current theory presumes that natural selection on vascular traits is controlled by a trade-off between efficiency and safety of hydraulic architecture. Hence, traits linked to efficiency, such as vessel diameter, should show biogeographic patterns; but critical tests of these predictions are rare, largely owing to confounding effects of environment, tree size and phylogeny. Using wood sampled from a phylogenetically constrained set of 28 Eucalyptus species, collected from a wide gradient of aridity across Australia, we show that hydraulic architecture reflects adaptive radiation of this genus in response to variation in climate. With increasing aridity, vessel diameters narrow, their frequency increases with a distribution that becomes gradually positively skewed and sapwood density increases while the theoretical hydraulic conductivity declines. Differences in these hydraulic traits appear largely genotypic in origin rather than environmentally plastic. Data reported here reflect long-term adaptation of hydraulic architecture to water availability. Rapidly changing climates, on the other hand, present significant challenges to the ability of eucalypts to adapt their vasculature.

Genetic variation in circadian regulation of nocturnal stomatal conductance enhances carbon assimilation and growth

Circadian resonance, whereby a plants endogenous rhythms are tuned to match environmental cues, has been repeatedly shown to be adaptive, although the underlying mechanisms remain elusive. Concomitantly, the adaptive value of nocturnal transpiration in C3 plants remains unknown because it occurs without carbon assimilation. These seemingly unrelated processes are interconnected because circadian regulation drives temporal patterns in nocturnal stomatal conductance, with maximum values occurring immediately before dawn for many species. We grew individuals of six Eucalyptus camaldulensis genotypes in naturally lit glasshouses and measured sunset, predawn and midday leaf gas exchange and whole-plant biomass production. We tested whether sunrise anticipation by the circadian clock and subsequent increases in genotype predawn stomatal conductance led to rapid stomatal opening upon illumination, ultimately affecting genotype differences in carbon assimilation and growth. We observed faster stomatal responses to light inputs at sunrise in genotypes with higher predawn stomatal conductance. Moreover, early morning and midday stomatal conductance and carbon assimilation, leaf area and total plant biomass were all positively correlated with predawn stomatal conductance across genotypes. Our results lead to the novel hypothesis that genotypic variation in the circadian-regulated capacity to anticipate sunrise could be an important factor underlying intraspecific variation in tree growth.

Industrial-age changes in atmospheric [CO2] and temperature differentially alter responses of faster- and slower-growing Eucalyptus seedlings to short-term drought.

Climate change may alter forest composition by differentially affecting the responses of faster- and slower-growing tree species to drought. However, the combined effects of rising atmospheric CO2 concentration ([CO2]) and temperature on drought responses of trees are poorly understood. Here, we examined interactive effects of temperature (ambient, ambient + °C) and [CO2] (290, 400 and 650mu;l l(-1)) on drought responses of Eucalyptus saligna Sm. (faster-growing) and E. sideroxylon A. Cunn. ex Woolls (slower-growing) seedlings. Drought was imposed via a controlled reduction in soil water over 1-2 weeks, re-watering seedlings when leaves visibly wilted. In ambient temperature, the effect of drought on the light-saturated net photosynthetic rate (Asat) in E. saligna decreased as [CO2] increased from pre-industrial to future concentrations, but rising [CO2] did not affect the response in Eucalyptus sideroxylon. In contrast, elevated temperature exacerbated the effect of drought in reducing Asat in both species. The drought response of Asat reflected changes in stomatal conductance (gs) associated with species and treatment differences in (i) utilization of soil moisture and (ii) leaf area ratio (leaf area per unit plant dry mass). Across [CO2] and temperature treatments, E. saligna wilted at higher soil water potentials compared with E. sideroxylon. Photosynthetic recovery from drought was 90% complete 2 days following re-watering across all species and treatments. Our results suggest that E. saligna (faster-growing) seedlings are more susceptible to drought than E. sideroxylon (slower-growing) seedlings. The greater susceptibility to drought of E. saligna reflected faster drawdown of soil moisture, associated with more leaf area and leaf area ratio, and the ability of E. sideroxylon to maintain higher gs at a given soil moisture. Inclusion of a pre-industrial [CO2] treatment allowed us to conclude that susceptibility of these species to short-term drought under past and future climates may be regulated by the same mechanisms. Further, the beneficial effects of rising [CO2] and deleterious effects of elevated temperature on seedling response to drought were generally offsetting, suggesting susceptibility of seedlings of these species to short-term drought in future climates that is similar to pre-industrial and current climate conditions.

Drought responses of two gymnosperm species with contrasting stomatal regulation strategies under elevated [CO2] and temperature

Future climate regimes characterized by rising [CO2], rising temperatures and associated droughts may differentially affect tree growth and physiology. However, the interactive effects of these three factors are complex because elevated [CO2] and elevated temperature may generate differential physiological responses during drought. To date, the interactive effects of elevated [CO2] and elevated temperature on drought-induced tree mortality remain poorly understood in gymnosperm species that differ in stomatal regulation strategies. Water relations and carbon dynamics were examined in two species with contrasting stomatal regulation strategies: Pinus radiata D. Don (relatively isohydric gymnosperm; regulating stomata to maintain leaf water potential above critical thresholds) and Callitris rhomboidea R. Br (relatively anisohydric gymnosperm; allowing leaf water potential to decline as the soil dries), to assess response to drought as a function of [CO2] and temperature. Both species were grown in two [CO2] (C(a) (ambient, 400 μl l(-1)) and C(e) (elevated, 640 μl l(-1))) and two temperature (T(a) (ambient) and T(e) (ambient +4 °C)) treatments in a sun-lit glasshouse under well-watered conditions. Drought plants were then exposed to a progressive drought until mortality. Prior to mortality, extensive xylem cavitation occurred in both species, but significant depletion of non-structural carbohydrates was not observed in either species. Te resulted in faster mortality in P. radiata, but it did not modify the time-to-mortality in C. rhomboidea. C(e) did not delay the time-to-mortality in either species under drought or T(e) treatments. In summary, elevated temperature (+4 °C) had greater influence than elevated [CO2] (+240 μl l(-1)) on drought responses of the two studied gymnosperm species, while stomatal regulation strategies did not generally affect the relative contributions of hydraulic failure and carbohydrate depletion to mortality under severe drought.

Leaf structural responses to pre-industrial, current and elevated atmospheric [CO2] and temperature affect leaf function in Eucalyptus sideroxylon

Leaf structure and chemistry both play critical roles in regulating photosynthesis. Yet, a key unresolved issue in climate change research is the role of changes in leaf structure in photosynthetic responses to temperature and atmospheric CO2 concentration ([CO2]), ranging from pre-industrial to future levels. We examined the interactive effects of [CO2] (290, 400 and 650μLL-1) and temperature (ambient, ambient +4°C) on leaf structural and chemical traits that regulate photosynthesis in Eucalyptus sideroxylon A.Cunn. ex Woolls. Rising [CO2] from pre-industrial to elevated levels increased light-saturated net photosynthetic rates (Asat), but reduced photosynthetic capacity (Amax). Changes in leaf N per unit area (Narea) and the number of palisade layers accounted for 56 and 14% of the variation in Amax, respectively, associated with changes in leaf mass per area. Elevated temperature increased stomatal frequency, but did not affect Amax. Further, rising [CO2] and temperature generally did not interactively affect leaf structure or function. These results suggest that leaf Narea and the number of palisade layers are the key chemical and structural factors regulating photosynthetic capacity of E. sideroxylon under rising [CO2], whereas the lack of photosynthetic responses to elevated temperature may reflect the limited effect of temperature on leaf structure and chemistry.

Examination of pre-industrial and future [CO2] reveals the temperature-dependent CO2 sensitivity of light energy partitioning at PSII in eucalypts

We grew faster-growing Eucalyptus saligna Sm. and slower-growing Eucalyptus sideroxylon A. Cunn ex Woolls tree seedlings in sunlit glasshouses at all combinations of 290 µL L–1 (pre-industrial), 400 µL L–1 (modern) or 650 µL L–1 (future) global atmospheric CO2 ([CO2]), and ambient or ambient + 4°C temperature. To assess photosynthetic performance, we simultaneously measured light-saturated CO2 assimilation (Asat) and chlorophyll fluorescence emission along with the capacity for photosynthetic O2 evolution and leaf pigment composition. Photosynthetic response to [CO2] was similar between species. Increasing [CO2] but not temperature increased Asat. The response of photosynthetic electron transport to [CO2] was temperature-dependent and manifested through adjustments in energy partitioning at PSII. Increasing [CO2] resulted in greater PSII operating efficiencies at the elevated temperature. We observed no associated acclimatory adjustments in the capacity for photosynthetic O2 evolution or changes in leaf chlorophyll content. Photoprotective energy dissipation responded to increasing [CO2] and temperature. Across species and treatments, increased energy partitioning to electron transport was always associated with decreased partitioning to energy dissipation. Our results suggest that in response to increasing [CO2] and temperature, E. saligna and E. sideroxylon meet increased demands for the products of electron transport via adjustments in energy partitioning, not through acclimation of the capacity for photosynthetic electron transport or light absorption.