Temperature responses of leaf respiration in light and darkness are similar and modulated by leaf development.

Our ability to predict temperature responses of leaf respiration in light and darkness (RL and RDk ) is essential to models of global carbon dynamics. While many models rely on constant thermal sensitivity (characterized by Q10 ), uncertainty remains as to whether Q10 of RL and RDk are actually similar. We measured short-term temperature responses of RL and RDk in immature and mature leaves of two evergreen tree species, Castanopsis carlesii and Ormosia henry in an open field. RL was estimated by the Kok method, the Yin method and a newly developed Kok-iterCc method. When estimated by the Yin and Kok-iterCc methods, RL and RDk had similar Q10 (c. 2.5). The Kok method overestimated both Q10 and the light inhibition of respiration. RL /RDk was not affected by leaf temperature. Acclimation of respiration in summer was associated with a decline in basal respiration but not in Q10 in both species, which was related to changes in leaf nitrogen content between seasons. Q10 of RL and RDk in mature leaves were 40% higher than in immature leaves. Our results suggest similar Q10 values can be used to model RL and RDk while leaf development-associated changes in Q10 require special consideration in future respiration models.

Advance methodological approaches for carbon stock estimation in forest ecosystems.

The forests are a key player in maintaining ecological balance on the earth. They not only conserve biodiversity, reduce soil erosion, and protect watersheds but also promote the above and below-ground ecosystem services. Forests are known as air cleaners on the planet and play a significant role in mitigating greenhouse gas (GHG) emissions into the atmosphere. As per programs launched in the Conference of Parties (COP) 26, there is a need to promote policies and programs to reduce the atmospheric carbon (C) through the forest ecosystem; it is because forests can capture the atmospheric CO2 for a long time and help to achieve the goals of net-zero emission CO2 on the earth. Therefore, there is an urgent need to know the advanced technological approaches for estimating C stock in forest ecosystems. Hence, the present article is aimed at providing a comprehensive protocol for the four C stock estimation approaches. An effort has also been made to compare these methods. This review suggests that tree allometry is the most common method used for the quantification of C stock, but this method has certain limitations. However, the review shows that accurate results can be produced by a combination of two or more methods. We have also analyzed the results of 42 research studies conducted for C stock assessment along with the factors determining the amount of C in different types of forests. The C stock in vegetation is affected by temporal and spatial variation, plantation age, land use, cropping pattern, management practices and elevation, etc. Nevertheless, the available results have a large degree of uncertainty mainly due to the limitations of the methods used. The review supports the conclusion that the uncertainty in C stock measurements can be addressed by the integration of the above-mentioned methods.

The crucial roles of mitochondria in supporting C4 photosynthesis.

C4 photosynthesis involves a series of biochemical and anatomical traits that significantly improve plant productivity under conditions that reduce the efficiency of C3 photosynthesis. We explore how evolution of the three classical biochemical types of C4 photosynthesis (NADP-ME, NAD-ME and PCK types) has affected the functions and properties of mitochondria. Mitochondria in C4 NAD-ME and PCK types play a direct role in decarboxylation of metabolites for C4 photosynthesis. Mitochondria in C4 PCK type also provide ATP for C4 metabolism, although this role for ATP provision is not seen in NAD-ME type. Such involvement has increased mitochondrial abundance/size and associated enzymatic capacity, led to changes in mitochondrial location and ultrastructure, and altered the role of mitochondria in cellular carbon metabolism in the NAD-ME and PCK types. By contrast, these changes in mitochondrial properties are absent in the C4 NADP-ME type and C3 leaves, where mitochondria play no direct role in photosynthesis. From an eco-physiological perspective, rates of leaf respiration in darkness vary considerably among C4 species but does not differ systematically among the three C4 types. This review outlines further mitochondrial research in key areas central to the engineering of the C4 pathway into C3 plants and to the understanding of variation in rates of C4 dark respiration.

Consistent diurnal pattern of leaf respiration in the light among contrasting species and climates.

Leaf daytime respiration (leaf respiration in the light, RL ) is often assumed to constitute a fixed fraction of leaf dark respiration (RD ) (i.e. a fixed light inhibition of respiration (RD )) and vary diurnally due to temperature fluctuations. These assumptions were tested by measuring RL , RD and the light inhibition of RD in the field at a constant temperature using the Kok method. Measurements were conducted diurnally on 21 different species: 13 deciduous, four evergreen and four herbaceous from humid continental and humid subtropical climates. RL and RD showed significant diurnal variations and the diurnal pattern differed in trajectory and magnitude between climates, but not between plant functional types (PFTs). The light inhibition of RD varied diurnally and differed between climates and in trajectory between PFTs. The results highlight the entrainment of leaf daytime respiration to the diurnal cycle and that time of day should be accounted for in studies seeking to examine the environmental and biological drivers of leaf daytime respiration.

Responses of leaf respiration to heatwaves.

Mitochondrial respiration (R) is central to plant physiology and responds dynamically to daily short-term temperature changes. In the longer-term, changes in energy demand and membrane fluidity can decrease leaf R at a common temperature and increase the temperature at which leaf R peaks (Tmax ). However, leaf R functionality is more susceptible to short-term heatwaves. Catalysis increases with rising leaf temperature, driving faster metabolism and leaf R demand, despite declines in photosynthesis restricting assimilate supply and growth. Proteins denature as temperatures increase further, adding to maintenance costs. Excessive heat also inactivates respiratory enzymes, with a concomitant limitation on the capacity of the R system. These competing push-and-pull factors are responsible for the diminishing acceleration in leaf R rate as temperature rises. Under extreme heat, membranes become overly fluid, and enzymes such as the cytochrome c oxidase are impaired. Such changes can lead to over-reduction of the energy system culminating in reactive oxygen species production. This ultimately leads to the total breakdown of leaf R, setting the limit of leaf survival. Understanding the heat stress responses of leaf R is imperative, given the continued rise in frequency and intensity of heatwaves and the importance of R for plant fitness and survival.

Modified Arrhenius Equation in Materials Science, Chemistry and Biology.

The Arrhenius plot (logarithmic plot vs. inverse temperature) is represented by a straight line if the Arrhenius equation holds. A curved Arrhenius plot (mostly concave) is usually described phenomenologically, often using polynomials of T or 1/T. Many modifications of the Arrhenius equation based on different models have also been published, which fit the experimental data better or worse. This paper proposes two solutions for the concave-curved Arrhenius plot. The first is based on consecutive A→B→C reaction with rate constants k1 ≪ k2 at higher temperatures and k1 ≫ k2 (or at least k1 > k2) at lower temperatures. The second is based on the substitution of the temperature T the by temperature difference T - T0 in the Arrhenius equation, where T0 is the maximum temperature at which the Arrheniusprocess under study does not yet occur.

Small tropical forest trees have a greater capacity to adjust carbon metabolism to long-term drought than large canopy trees.

The response of small understory trees to long-term drought is vital in determining the future composition, carbon stocks and dynamics of tropical forests. Long-term drought is, however, also likely to expose understory trees to increased light availability driven by drought-induced mortality. Relatively little is known about the potential for understory trees to adjust their physiology to both decreasing water and increasing light availability. We analysed data on maximum photosynthetic capacity (Jmax , Vcmax ), leaf respiration (Rleaf ), leaf mass per area (LMA), leaf thickness and leaf nitrogen and phosphorus concentrations from 66 small trees across 12 common genera at the world's longest running tropical rainfall exclusion experiment and compared responses to those from 61 surviving canopy trees. Small trees increased Jmax , Vcmax , Rleaf and LMA (71, 29, 32, 15% respectively) in response to the drought treatment, but leaf thickness and leaf nutrient concentrations did not change. Small trees were significantly more responsive than large canopy trees to the drought treatment, suggesting greater phenotypic plasticity and resilience to prolonged drought, although differences among taxa were observed. Our results highlight that small tropical trees have greater capacity to respond to ecosystem level changes and have the potential to regenerate resilient forests following future droughts.

Macromolecular rate theory (MMRT) provides a thermodynamics rationale to underpin the convergent temperature response in plant leaf respiration.

Temperature is a crucial factor in determining the rates of ecosystem processes, for example, leaf respiration (R) - the flux of plant respired CO2 from leaves to the atmosphere. Generally, R increases exponentially with temperature and formulations such as the Arrhenius equation are widely used in earth system models. However, experimental observations have shown a consequential and consistent departure from an exponential increase in R. What are the principles that underlie these observed patterns? Here, we demonstrate that macromolecular rate theory (MMRT), based on transition state theory (TST) for enzyme-catalyzed kinetics, provides a thermodynamic explanation for the observed departure and the convergent temperature response of R using a global database. Three meaningful parameters emerge from MMRT analysis: the temperature at which the rate of respiration would theoretically reach a maximum (the optimum temperature, Topt ), the temperature at which the respiration rate is most sensitive to changes in temperature (the inflection temperature, Tinf ) and the overall curvature of the log(rate) versus temperature plot (the change in heat capacity for the system, ΔCP‡). On average, the highest potential enzyme-catalyzed rates of respiratory enzymes for R are predicted to occur at 67.0 ± 1.2°C and the maximum temperature sensitivity at 41.4 ± 0.7°C from MMRT. The average curvature (average negative ΔCP‡) was -1.2 ± 0.1 kJ mol-1  K-1 . Interestingly, Topt , Tinf and ΔCP‡ appear insignificantly different across biomes and plant functional types, suggesting that thermal response of respiratory enzymes in leaves could be conserved. The derived parameters from MMRT can serve as thermal traits for plant leaves that represent the collective temperature response of metabolic respiratory enzymes and could be useful to understand regulations of R under a warmer climate. MMRT extends the classic TST to enzyme-catalyzed reactions and provides an accurate and mechanistic model for the short-term temperature response of R around the globe.

Photosynthetic capacity and leaf nitrogen decline along a controlled climate gradient in provenances of two widely distributed Eucalyptus species.

Climate is an important factor limiting tree distributions and adaptation to different thermal environments may influence how tree populations respond to climate warming. Given the current rate of warming, it has been hypothesized that tree populations in warmer, more thermally stable climates may have limited capacity to respond physiologically to warming compared to populations from cooler, more seasonal climates. We determined in a controlled environment how several provenances of widely distributed Eucalyptus tereticornis and E. grandis adjusted their photosynthetic capacity to +3.5°C warming along their native distribution range (~16-38°S) and whether climate of seed origin of the provenances influenced their response to different growth temperatures. We also tested how temperature optima (Topt ) of photosynthesis and Jmax responded to higher growth temperatures. Our results showed increased photosynthesis rates at a standardized temperature with warming in temperate provenances, while rates in tropical provenances were reduced by about 40% compared to their temperate counterparts. Temperature optima of photosynthesis increased as provenances were exposed to warmer growth temperatures. Both species had ~30% reduced photosynthetic capacity in tropical and subtropical provenances related to reduced leaf nitrogen and leaf Rubisco content compared to temperate provenances. Tropical provenances operated closer to their thermal optimum and came within 3% of the Topt of Jmax during the daily temperature maxima. Hence, further warming may negatively affect C uptake and tree growth in warmer climates, whereas eucalypts in cooler climates may benefit from moderate warming.

Scaling leaf respiration with nitrogen and phosphorus in tropical forests across two continents.

Leaf dark respiration (Rdark ) represents an important component controlling the carbon balance in tropical forests. Here, we test how nitrogen (N) and phosphorus (P) affect Rdark and its relationship with photosynthesis using three widely separated tropical forests which differ in soil fertility. Rdark was measured on 431 rainforest canopy trees, from 182 species, in French Guiana, Peru and Australia. The variation in Rdark was examined in relation to leaf N and P content, leaf structure and maximum photosynthetic rates at ambient and saturating atmospheric CO2 concentration. We found that the site with the lowest fertility (French Guiana) exhibited greater rates of Rdark per unit leaf N, P and photosynthesis. The data from Australia, for which there were no phylogenetic overlaps with the samples from the South American sites, yielded the most distinct relationships of Rdark with the measured leaf traits. Our data indicate that no single universal scaling relationship accounts for variation in Rdark across this large biogeographical space. Variability between sites in the absolute rates of Rdark and the Rdark  : photosynthesis ratio were driven by variations in N- and P-use efficiency, which were related to both taxonomic and environmental variability.

Inside out: efflux of carbon dioxide from leaves represents more than leaf metabolism.

High concentrations of inorganic carbon in the xylem, produced from root, stem, and branch respiration, travel via the transpiration stream and eventually exit the plant through distant tissues as CO2. Unlike previous studies that focused on the efflux of CO2 from roots and woody tissues, we focus on efflux from leaves and the potential effect on leaf respiration measurements. We labeled transported inorganic carbon, spanning reported xylem concentrations, with 13C and then manipulated transpiration rates in the dark in order to vary the rates of inorganic carbon supply to cut leaves from Brassica napus and Populus deltoides. We used tunable diode laser absorbance spectroscopy to directly measure the rate of gross 13CO2 efflux, derived from inorganic carbon supplied from outside of the leaf, relative to gross 12CO2 efflux generated from leaf cells. These experiemnts showed that 13CO2 efflux was dependent upon the rate of inorganic carbon supply to the leaf and the rate of transpiration. Our data show that the gross leaf efflux of xylem-transported CO2 is likely small in the dark when rates of transpiration are low. However, gross leaf efflux of xylem-transported CO2 could approach half the rate of leaf respiration in the light when transpiration rates and branch inorganic carbon concentrations are high, irrespective of the grossly different petiole morphologies in our experiment.

A test of the 'one-point method' for estimating maximum carboxylation capacity from field-measured, light-saturated photosynthesis.

Simulations of photosynthesis by terrestrial biosphere models typically need a specification of the maximum carboxylation rate (Vcmax ). Estimating this parameter using A-Ci curves (net photosynthesis, A, vs intercellular CO2 concentration, Ci ) is laborious, which limits availability of Vcmax data. However, many multispecies field datasets include net photosynthetic rate at saturating irradiance and at ambient atmospheric CO2 concentration (Asat ) measurements, from which Vcmax can be extracted using a 'one-point method'. We used a global dataset of A-Ci curves (564 species from 46 field sites, covering a range of plant functional types) to test the validity of an alternative approach to estimate Vcmax from Asat via this 'one-point method'. If leaf respiration during the day (Rday ) is known exactly, Vcmax can be estimated with an r(2) value of 0.98 and a root-mean-squared error (RMSE) of 8.19 µmol m(-2) s(-1) . However, Rday typically must be estimated. Estimating Rday as 1.5% of Vcmax, we found that Vcmax could be estimated with an r(2) of 0.95 and an RMSE of 17.1 µmol m(-2) s(-1) . The one-point method provides a robust means to expand current databases of field-measured Vcmax , giving new potential to improve vegetation models and quantify the environmental drivers of Vcmax variation.

Leaf gas films, underwater photosynthesis and plant species distributions in a flood gradient.

Traits for survival during flooding of terrestrial plants include stimulation or inhibition of shoot elongation, aerenchyma formation and efficient gas exchange. Leaf gas films form on superhydrophobic cuticles during submergence and enhance underwater gas exchange. The main hypothesis tested was that the presence of leaf gas films influences the distribution of plant species along a natural flood gradient. We conducted laboratory experiments and field observations on species distributed along a natural flood gradient. We measured presence or absence of leaf gas films and specific leaf area of 95 species. We also measured, gas film retention time during submergence and underwater net photosynthesis and dark respiration of 25 target species. The presence of a leaf gas film was inversely correlated to flood frequency and duration and reached a maximum value of 80% of the species in the rarely flooded locations. This relationship was primarily driven by grasses that all, independently of their field location along the flood gradient, possess gas films when submerged. Although the present study and earlier experiments have shown that leaf gas films enhance gas exchange of submerged plants, the ability of species to form leaf gas films did not show the hypothesized relationship with species composition along the flood gradient.

Developmental stage specificity of transcriptional, biochemical and CO2 efflux responses of leaf dark respiration to growth of Arabidopsis thaliana at elevated [CO2].

Plant respiration responses to elevated growth [CO(2)] are key uncertainties in predicting future crop and ecosystem function. In particular, the effects of elevated growth [CO(2)] on respiration over leaf development are poorly understood. This study tested the prediction that, due to greater whole plant photoassimilate availability and growth, elevated [CO(2)] induces transcriptional reprogramming and a stimulation of nighttime respiration in leaf primordia, expanding leaves and mature leaves of Arabidopsis thaliana. In primordia, elevated [CO(2)] altered transcript abundance, but not for genes encoding respiratory proteins. In expanding leaves, elevated [CO(2)] induced greater glucose content and transcript abundance for some respiratory genes, but did not alter respiratory CO(2) efflux. In mature leaves, elevated [CO(2)] led to greater glucose, sucrose and starch content, plus greater transcript abundance for many components of the respiratory pathway, and greater respiratory CO(2) efflux. Therefore, growth at elevated [CO(2)] stimulated dark respiration only after leaves transitioned from carbon sinks into carbon sources. This coincided with greater photoassimilate production by mature leaves under elevated [CO(2)] and peak respiratory transcriptional responses. It remains to be determined if biochemical and transcriptional responses to elevated [CO(2)] in primordial and expanding leaves are essential prerequisites for subsequent alterations of respiratory metabolism in mature leaves.

The mechanism of improved aeration due to gas films on leaves of submerged rice.

Some terrestrial wetland plants, such as rice, have super-hydrophobic leaf surfaces which retain a gas film when submerged. O2 movement through the diffusive boundary layer (DBL) of floodwater, gas film and stomata into leaf mesophyll was explored by means of a reaction-diffusion model that was solved in a three-dimensional leaf anatomy model. The anatomy and dark respiration of leaves of rice (Oryza sativa L.) were measured and used to compute O2 fluxes and partial pressure of O2 (pO2 ) in the DBL, gas film and leaf when submerged. The effects of floodwater pO2 , DBL thickness, cuticle permeability, presence of gas film and stomatal opening were explored. Under O2 -limiting conditions of the bulk water (pO2 < 10 kPa), the gas film significantly increases the O2 flux into submerged leaves regardless of whether stomata are fully or partly open. With a gas film, tissue pO2 substantially increases, even for the slightest stomatal opening, but not when stomata are completely closed. The effect of gas films increases with decreasing cuticle permeability. O2 flux and tissue pO2 decrease with increasing DBL thickness. The present modelling analysis provides a mechanistic understanding of how leaf gas films facilitate O2 entry into submerged plants.

Balance between carbon gain and loss in warmer environments: impacts on photosynthesis and leaf respiration in four temperate tree species.

The temperature sensitivities of photosynthesis and respiration remain a key uncertainty in predicting how forests will respond to climate warming. We grew seedlings of four temperate tree species, including Betula platyphylla, Fraxinus mandshurica, Juglans mandshurica and Tilia amurensis, at three temperature regimes (ambient, +2 °C, and +4 °C in daytime air temperature). We investigated net photosynthesis (Anet25), maximum rate of RuBP-carboxylation (Vcmax25) and RuBP-regeneration (Jmax25), stomatal conductance (gs25), mesophyll conductance (gm25), and leaf respiration (Rleaf) in dark (Rdark25) and in light (Rlight25) at 25 °C in all species. Additionally, we examined the temperature sensitivities of Anet, Vcmax, Jmax, Rdark and Rlight in F. mandshurica. Our findings showed that the warming-induced decreases in Anet25, Vcmax25 and Jmax25 were more prevalent in the late-successional species T. amurensis. Warming had negative impacts on gs25 in all species. Overall, Anet25 was positively correlated with Vcmax25 and Jmax25 across all growth temperatures. However, a positive correlation between Anet25 and gs25 was observed only under warming conditions, and gs25 was negatively associated with vapor pressure deficit. This implies that the vapor pressure deficit-induced decrease in gs25 was responsible for the decline in Anet25 at higher temperatures. The optimum temperature of Anet in F. mandshurica increased by 0.59 °C per 1.0 °C rise in growth temperature. While +2 °C elevated the thermal optima of Jmax, it did not affect the other temperature sensitivity parameters of Vcmax and Jmax. Rdark25 was not affected by warming in any species, and Rlight25 was stimulated in T. amurensis. The temperature response curves of Rdark and Rlight in F. mandshurica were not altered by warming, implying a lack of thermal acclimation. The ratios of Rdark25 and Rlight25 to Anet25 and Vcmax25 in T. amurensis increased with warming. These results suggest that Anet and Rleaf did not acclimate to warming synchronously in these temperate tree species.

Respiration in light of evergreen and deciduous woody species and its links to the leaf economic spectrum.

Leaf respiration in the light (Rlight) is crucial for understanding the net CO2 exchange of individual plants and entire ecosystems. However, Rlight is poorly quantified and rarely discussed in the context of the leaf economic spectrum (LES), especially among woody species differing in plant functional types (PFTs) (e.g., evergreen vs. deciduous species). To address this gap in our knowledge, Rlight, respiration in the dark (Rdark), light-saturated photosynthetic rates (Asat), leaf dry mass per unit area (LMA), leaf nitrogen (N) and phosphorus (P) concentrations, and maximum carboxylation (Vcmax) and electron transport rates (Jmax) of 54 representative subtropical woody evergreen and deciduous species were measured. With the exception of LMA, the parameters quantified in this study were significantly higher in deciduous species than in evergreen species. The degree of light inhibition did not significantly differ between evergreen (52%) and deciduous (50%) species. Rlight was significantly correlated with LES traits such as Asat, Rdark, LMA, N and P. The Rlight vs. Rdark and N relationships shared common slopes between evergreen and deciduous species, but significantly differed in their y-intercepts, in which the rates of Rlight were slower or faster for any given Rdark or N in deciduous species, respectively. A model for Rlight based on three traits (i.e., Rdark, LMA and P) had an explanatory power of 84.9%. These results show that there is a link between Rlight and the LES, and highlight that PFTs is an important factor in affecting Rlight and the relationships of Rlight with Rdark and N. Thus, this study provides information that can improve the next generation of terrestrial biosphere models (TBMs).

Reduction of molecular gas diffusion through gaskets in leaf gas exchange cuvettes by leaf-mediated pores.

There is an ongoing debate on how to correct leaf gas exchange measurements for the unavoidable diffusion leakage that occurs when measurements are done in non-ambient CO2 concentrations. In this study, we present a theory on how the CO2 diffusion gradient over the gasket is affected by leaf-mediated pores (LMP) and how LMP reduce diffusive exchange across the gaskets. Recent discussions have so far neglected the processes in the quasi-laminar boundary layer around the gasket. Counter intuitively, LMP reduce the leakage through gaskets, which can be explained by assuming that the boundary layer at the exterior of the cuvette is enriched with air from the inside of the cuvette. The effect can thus be reduced by reducing the boundary layer thickness. The theory clarifies conflicting results from earlier studies. We developed leaf adaptor frames that eliminate LMP during measurements on delicate plant material such as grass leaves with circular cross section, and the effectiveness is shown with respiration measurements on a harp of Deschampsia flexuosa leaves. We conclude that the best solution for measurements with portable photosynthesis systems is to avoid LMP rather than trying to correct for the effects.

Carbon isotope composition of respired CO2 in woody stems and leafy shoots of three tree species along the growing season: physiological drivers for respiratory fractionation.

The carbon isotope composition of respired CO2 (δ13CR) and bulk organic matter (δ13CB) of various plant compartments informs about the isotopic fractionation and substrate of respiratory processes, which are crucial to advance the understanding of carbon allocation in plants. Nevertheless, the variation across organs, species and seasons remains poorly understood. Cavity Ring-Down Laser Spectroscopy was applied to measure δ13CR in leafy shoots and woody stems of maple (Acer platanoides L.), oak (Quercus robur L.) and cedar (Thuja occidentalis L.) trees during spring and late summer. Photosynthesis, respiration, growth and non-structural carbohydrates were measured in parallel to evaluate potential drivers for respiratory fractionation. The CO2 respired by maple and oak shoots was 13C-enriched relative to δ13CB during spring, but not late summer or in the stem. In cedar, δ13CR did not vary significantly throughout organs and seasons, with respired CO2 being 13C-depleted relative to δ13CB. Shoot δ13CR was positively related to leaf starch concentration in maple, while stem δ13CR was inversely related to stem growth. These relations were not significant for oak or cedar. The variability in δ13CR suggests (i) different contributions of respiratory pathways between organs and (ii) seasonality in the respiratory substrate and constitutive compounds for wood formation in deciduous species, less apparent in evergreen cedar, whose respiratory metabolism might be less variable.