Differential coordination of stomatal conductance,mesophyll conductance,and leaf hydraulic conductance in response to changing light across species

Stomatal conductance (g_s) and mesophyll conductance (g_m) represent major constraints to photosynthetic rate (A), and these traits are expected to coordinate with leaf hydraulic conductance (K_leaf) across species, under both steady‐state and dynamic conditions. However, empirical information about their coordination is scarce. In this study, K_leaf, gas exchange, stomatal kinetics, and leaf anatomy in 10 species including ferns, gymnosperms, and angiosperms were investigated to elucidate the correlation of H₂O and CO₂ diffusion inside leaves under varying light conditions. Gas exchange, K_leaf, and anatomical traits varied widely across species. Under light‐saturated conditions, the A, g_s, g_m, and K_leaf were strongly correlated across species. However, the response patterns of A, g_s, g_m, and K_leaf to varying light intensities were highly species dependent. Moreover, stomatal opening upon light exposure of dark‐adapted leaves in the studied ferns and gymnosperms was generally faster than in the angiosperms; however, stomatal closing in light‐adapted leaves after darkening was faster in angiosperms. The present results show that there is a large variability in the coordination of leaf hydraulic and gas exchange parameters across terrestrial plant species, as well as in their responses to changing light.     

Coupled response of stomatal and mesophyll conductance to light enhances photosynthesis of shade leaves under sunflecks

Light gradients within tree canopies play a major role in the distribution of plant resources that define the photosynthetic capacity of sun and shade leaves. However, the biochemical and diffusional constraints on gas exchange in sun and shade leaves in response to light remain poorly quantified, but critical for predicting canopy carbon and water exchange. To investigate the CO₂ diffusion pathway of sun and shade leaves, leaf gas exchange was coupled with concurrent measurements of carbon isotope discrimination to measure net leaf photosynthesis (A_n), stomatal conductance (g_s) and mesophyll conductance (g_m) in Eucalyptus tereticornis trees grown in climate controlled whole‐tree chambers. Compared to sun leaves, shade leaves had lower A_n, g_m, leaf nitrogen and photosynthetic capacity (A_max) but g_s was similar. When light intensity was temporarily increased for shade leaves to match that of sun leaves, both g_s and g_m increased, and A_n increased to values greater than sun leaves. We show that dynamic physiological responses of shade leaves to altered light environments have implications for up‐scaling leaf level measurements and predicting whole canopy carbon gain. Despite exhibiting reduced photosynthetic capacity, the rapid up‐regulation of g_m with increased light enables shade leaves to respond quickly to sunflecks.     

The role of aquaporins and membrane damage in chilling and hydrogen peroxide induced changes in the hydraulic conductance of maize roots

When chilling-sensitive plants are chilled, root hydraulic conductance (L(o)) declines precipitously; L(o) also declines in chilling-tolerant plants, but it subsequently recovers, whereas in chilling-sensitive plants it does not. As a result, the chilling-sensitive plants dry out and may die. Using a chilling-sensitive and a chilling-tolerant maize genotype we investigated the effect of chilling on L(o), and its relationship to osmotic water permeability of isolated root cortex protoplasts, aquaporin gene expression, aquaporin abundance, and aquaporin phosphorylation, hydrogen peroxide (H(2)O(2)) accumulation in the roots and electrolyte leakage from the roots. Because chilling can cause H(2)O(2) accumulation we also determined the effects of a short H(2)O(2) treatment of the roots and examined the same parameters. We conclude from these studies that the recovery of L(o) during chilling in the chilling-tolerant genotype is made possible by avoiding or repairing membrane damage and by a greater abundance and/or activity of aquaporins. The same changes in aquaporins take place in the chilling-sensitive genotype, but we postulate that membrane damage prevents the L(o) recovery. It appears that the aquaporin response is necessary but not sufficient to respond to chilling injury. The plant must also be able to avoid the oxidative damage that accompanies chilling.     

The rapid light response of leaf hydraulic conductance: new evidence from two experimental methods

Previous studies have shown a rapid enhancement in leaf hydraulic conductance (K(leaf)) from low to high irradiance (from <10 to >1000 micromol photons m(-2) s(-1)), using the high-pressure flow meter (HPFM), for 7 of 14 tested woody species. However, theoretical suggestions have been made that this response might arise as an artifact of the HPFM. We tested the K(leaf) light response for six evergreen species using refined versions of the rehydration kinetics method (RKM) and the evaporative flux method (EFM). We found new evidence for a rapid, 60% to 100% increase in K(leaf) from low to high irradiance for three species. In the RKM, the leaf rehydration time constant declined by up to 70% under high irradiance relative to darkness. In the EFM, under higher irradiance, the flow rate increased disproportionately to the water potential gradient. Combining our data with those of previous studies, we found that heterobaric species, i.e. those with bundle sheath extensions (BSEs) showed a twofold greater K(leaf) light response on average than homobaric species, i.e. those without BSEs. We suggest further research to characterize this substantial dynamic at the nexus of plant light- and water-relations.     

Growth environment determines light sensitivity of shoot hydraulic conductance

Acclimation of light sensitivity of hydraulic conductance of shoots of silver birch (Betula pendula) and hybrid aspen (Populus × wettsteinii) to growth environments with three different air humidities was studied. Hydraulic conductance of shoots kept for 1–2 h in darkness (D) or in light (L) was measured by the pressure chamber method, and light sensitivity was defined as a significant difference between D and L shoots. Light sensitivity of shoots grown in three different air humidities was found to vary. Amongst shoots grown in current natural air, only the hydraulic conductance of the whole shoot and that of the leaf blades of birch upper foliage were significantly light sensitive. Amongst shoots grown in decreased air humidity, hydraulic conductance of the whole shoot, the leaf blades, and the stem and petioles of birch upper foliage, the conductance of the whole shoot and the leaf blades of birch lower foliage, and the conductance of the whole shoot of aspen upper foliage were light sensitive. None of the shoots grown in increased air humidity were significantly light sensitive. We predict that light sensitivity will become more widespread among species in regions where air humidity decreases as a result of global climate change, and vice versa. Low white light always caused the same increase in hydraulic conductance as high white light, and blue and white light always caused an increase in conductance about two times greater than red light, indicating that growth environment did not markedly modify the mechanism of light sensitivity.     

Rapid shoot‐to‐root signalling regulates root hydraulic conductance via aquaporins

We investigated how root hydraulic conductance (normalized to root dry weight, L_o) is regulated by the shoot. Shoot topping (about 30% reduction in leaf area) reduced L_o of grapevine (Vitis vinifera L.), soybean (Glycine max L.) and maize (Zea mays L.) by 50 to 60%. More detailed investigations with soybean and grapevine showed that the reduction in L_o was not correlated with the reduction in leaf area, and shading or cutting single leaves had a similar effect. Percentage reduction in L_o was largest when initial L_o was high in soybean. Inhibition of L_o by weak acid (low pH) was smaller after shoot damage or leaf shading. The half time of reduction in L_o was approximately 5 min after total shoot decapitation. These characteristics indicate involvement of aquaporins. We excluded phloem‐borne signals and auxin‐mediated signals. Xylem‐mediated hydraulic signals are possible since turgor rapidly decreased within root cortex cells after shoot topping. There was a significant reduction in the expression of several aquaporins in the plasma membrane intrinsic protein (PIP) family of both grapevine and soybean. In soybean, there was a five‐ to 10‐fold reduction in GmPIP1;6 expression over 0.5–1 h which was sustained over the period of reduced L_o.     

Oscillations in stomatal conductance of hybrid poplar leaves in the light and dark

Cycling of stomatal conductance in three hybrid poplar ( Populus sp.) cultivars was observed under a variety of conditions. Illumination of plants kept previously in the dark induced very large oscillations with a period of about 40 min and large oscillations with a shorter period (< 10 min) were superimposed on the longer cycles. During these oscillations, large changes in conductance could occur very rapidly (1.0 cm s⁻¹ in 3 min). Plants in constant light also displayed both long and short term cycles in conductance, but these were smaller in amplitude than those induced by sudden illumination. Stomatal oscillations were also observed in darkness and after darkening of previously illuminated plants. These oscillations had shorter (< 30 min) and less regular periods than those observed in the light. Such cycling in the dark is rare. Cycling of the two leaf surfaces was sometimes in synchrony in the light, and more so after a perturbation. Little synchrony between the two surfaces was observed in the dark. Stomatal movements of different leaves on a plant were usually relatively independent. Transient stomatal opening occurred following leaf excision in the light or dark, and often after sudden darkening of intact leaves. Also, stomata of intact leaves sometimes transiently closed following illumination.     

The contribution of photosynthesis to the red light response of stomatal conductance

To determine the contribution of photosynthesis on stomatal conductance, we contrasted the stomatal red light response of wild-type tobacco (Nicotiana tabacum 'W38') with that of plants impaired in photosynthesis by antisense reductions in the content of either cytochrome b(6)f complex (anti-b/f plants) or Rubisco (anti-SSU plants). Both transgenic genotypes showed a lowered content of the antisense target proteins in guard cells as well as in the mesophyll. In the anti-b/f plants, CO(2) assimilation rates were proportional to leaf cytochrome b(6)f content, but there was little effect on stomatal conductance and the rate of stomatal opening. To compare the relationship between photosynthesis and stomatal conductance, wild-type plants and anti-SSU plants were grown at 30 and 300 micromol photon m(-2) s(-1) irradiance (low light and medium light [ML], respectively). Growth in ML increased CO(2) assimilation rates and stomatal conductance in both genotypes. Despite the significantly lower CO(2) assimilation rate in the anti-SSU plants, the differences in stomatal conductance between the genotypes were nonsignificant at either growth irradiance. Irrespective of plant genotype, stomatal density in the two leaf surfaces was 2-fold higher in ML-grown plants than in low-light-grown plants and conductance normalized to stomatal density was unaffected by growth irradiance. We conclude that the red light response of stomatal conductance is independent of the concurrent photosynthetic rate of the guard cells or of that of the underlying mesophyll. Furthermore, we suggest that the correlation of photosynthetic capacity and stomatal conductance observed under different light environments is caused by signals largely independent of photosynthesis.     

Rapid variations of mesophyll conductance in response to changes in CO2 concentration around leaves

The effects of short-term (minutes) variations of CO2 concentration on mesophyll conductance to CO2 (gm) were evaluated in six different C3 species by simultaneous measurements of gas exchange, chlorophyll fluorescence, online carbon isotope discrimination and a novel curve-fitting method. Depending on the species, gm varied from five- to ninefold, along the range of sub-stomatal CO2 concentrations typically used in photosynthesis CO2-response curves (AN)-Ci curves; where AN is the net photosynthetic flux and Ci is the CO2 concentrations in the sub-stomatal cavity), that is, 50 to 1500 micromol CO2 mol(-1) air. Although the pattern was species-dependent, gm strongly declined at high Ci, where photosynthesis was not limited by CO2, but by regeneration of ribulose-1,5-bisphosphate or triose phosphate utilization. Moreover, these changes on gm were found to be totally independent of the velocity and direction of the Ci changes. The response of gm to Ci resembled that of stomatal conductance (gs), but kinetic experiments suggested that the response of gm was actually faster than that of gs. Transgenic tobacco plants differing in the amounts of aquaporin NtAQP1 showed different slopes of the gm-Ci response, suggesting a possible role for aquaporins in mediating CO2 responsiveness of gm. The importance of these findings is discussed in terms of their effects on parameterization of AN-Ci curves.     

Isoprenoid emissions,photosynthesis and mesophyll diffusion conductance in response to blue light

The effects of blue light (BL) on leaf gas exchange of Populus × canadensis, a strong isoprene emitter, and Quercus ilex and Citrus reticulata, two monoterpene emitters with respectively small and large storage pools for monoterpenes, were studied. Leaves were initially exposed to a saturating photosynthetic photon flux density (PPFD) of white light (WL), which was then progressively reduced to perform WL-response curves. Leaves acclimated to saturating WL were then quickly exposed to equivalent BL levels to perform BL-response curves. Blue light did not significantly affect photosynthetic parameters in the light-limited portion of the PPFD-response curves in both P. × canadensis and Q. ilex. Whereas photosynthesis (A), stomatal conductance (g_s), and mesophyll conductance (g_m) were significantly decreased at high PPFDs of BL. A was similarly inhibited by BL in C. reticulata, but there was no significant effect of light quality on g_s. Overall these results show that the negative effect of BL on photosynthesis is widespread in tree species with different leaf characteristics, and that this involves coordinated reductions in g_s and g_m. BL negatively affected isoprene emission and, to a lesser extent monoterpene emissions, in concert with photosynthetic inhibition. Interesting, both isoprene and monoterpene emissions were shown to be inversely dependent upon intercellular [CO₂]. These results indicate that a change in light spectral quality, which can vary during the day, between days and within seasons, can alter photosynthesis and isoprenoid emissions, depending on the PPFD intensity. Such effects should be strongly considered in photosynthesis and volatile isoprenoid emission models.     

The expression pattern of plasma membrane aquaporins in maize leaf highlights their role in hydraulic regulation

Leaves are key organs for evaporation and photosynthesis and play a crucial role in plant growth and development. In order to function properly, they need to maintain a balanced water content. Water movement through a leaf occurs by a combination of different pathways: water can follow an apoplastic route through the cell wall or a cell-to-cell route via the symplastic and transcellular paths. As aquaporins (AQPs) play an important role in regulating transcellular water flow and CO(2) conductance, studies on AQP mRNA and protein expression in leaves are essential to better understand their role in these physiological processes. Here, we quantified and localized the expression of Zea mays plasma membrane aquaporins (ZmPIPs, plasma membrane intrinsic proteins) in the leaf using quantitative RT-PCR and immunodetection. All ZmPIP genes except ZmPIP2;7 were expressed in leaves. Expression was found to be dependent on the developmental stage of the leaf tissue, with, in general, an increase in expression at the end of the elongation zone and a decrease in mature leaf tissue. These data correlated with the cell water permeability, as determined using a protoplast swelling assay. The diurnal expression of ZmPIPs was also investigated and expression was found to be higher during the first hours of the light period than at night. Immunocytochemical localization of four ZmPIP isoforms indicated that they are involved in leaf radial water movement, in particular in vascular bundles and the mesophyll.     

Co-ordination of hydraulic and stomatal conductances across light qualities in cucumber leaves

Long-term effects of light quality on leaf hydraulic conductance (K(leaf)) and stomatal conductance (g(s)) were studied in cucumber, and their joint impact on leaf photosynthesis in response to osmotic-induced water stress was assessed. Plants were grown under low intensity monochromatic red (R, 640 nm), blue (B, 420 nm) or combined red and blue (R:B, 70:30) light. K(leaf) and g(s) were much lower in leaves that developed without blue light. Differences in g(s) were caused by differences in stomatal aperture and stomatal density, of which the latter was largely due to differences in epidermal cell size and hardly due to stomatal development. Net photosynthesis (A(N)) was lowest in R-, intermediate in B-, and highest in RB- grown leaves. The low A(N) in R-grown leaves correlated with a low leaf internal CO(2) concentration and reduced PSII operating efficiency. In response to osmotic stress, all leaves showed similar degrees of stomatal closure, but the reduction in A(N) was larger in R- than in B- and RB-grown leaves. This was probably due to damage of the photosynthetic apparatus, which only occurred in R-grown leaves. The present study shows the co-ordination of K(leaf) and g(s) across different light qualities, while the presence of blue in the light spectrum seems to drive both K(leaf) and g(s) towards high, sun-type leaf values, as was previously reported for maximal photosynthetic capacity and leaf morphology. The present results suggest the involvement of blue light receptors in the usually harmonized development of leaf characteristics related to water relations and photosynthesis under different light environments.     

The chloroplast avoidance response decreases internal conductance to CO2 diffusion in Arabidopsis thaliana leaves

The relationship between chloroplast arrangement and diffusion of CO(2) from substomatal cavities to the chloroplast stroma was investigated in Arabidopsis thaliana. Chloroplast position was manipulated by varying the amount of blue light and by cytochalasin D (CytD) treatment. We also investigated two chloroplast positioning mutants. Chloroplast arrangement was assessed by the surface area of chloroplasts adjacent to intercellular airspaces (S(c)). Although it has been previously shown that long-term acclimation to high light is linked with a large S(c), we found that the short-term chloroplast avoidance response reduces S(c). This effect was not apparent in the blue-light-insensitive phot2 mutant, which did not show the avoidance response. As expected, the smaller S(c) induced by the avoidance response was coupled to a similar decrease in internal conductance. This reduction in internal conductance resulted in an increased limitation of the rate of photosynthesis. The limiting effect of S(c) on internal conductance and photosynthesis was also shown in chup1, a mutant with a constant small S(c) as the result of an unusual chloroplast arrangement. We conclude that chloroplast movements in A. thaliana can rapidly alter leaf morphological parameters, and this has significant consequences for the diffusion of CO(2) through the mesophyll.     

Dorsoventral photosynthetic asymmetry of tobacco leaves in response to direct and diffuse light

Journal of Plant Research - Plants can change leaf forms, adjusting light conditions on their adaxial and abaxial surfaces, to adapt to light environments and enhance their light use efficiencies....     

Use of the response of photosynthesis to oxygen to estimate mesophyll conductance to carbon dioxide in water-stressed soybean leaves

Methods of estimating the mesophyll conductance (g_m) to the movement of CO₂ from the substomatal airspace to the site of fixation are expensive or rely upon numerous assumptions. It is proposed that, for C₃ species, measurement of the response of photosynthesis to [O₂] at limiting [CO₂], combined with a standard biochemical model of photosynthesis, can provide an estimate of g_m. This method was used to determine whether g_m changed with [CO₂] and with water stress in soybean leaves. The value of g_m estimated using the O₂ response method agreed with values obtained using other methods. The g_m was unchanged over the tested range of substomatal [CO₂]. Water stress, which decreased stomatal conductance (g_s) by about 80%, did not affect g_m, while the model parameter V_Cmax was reduced by about 25%. Leaves with g_s reduced by about 90% had g_m values reduced by about 50%, while V_Cmax was reduced by about 64%. It is concluded that g_m in C₃ species can be conveniently estimated using the response of photosynthesis to [O₂] at limiting [CO₂], and that g_m in soybean was much less sensitive to water stress than g_s, and was somewhat less sensitive to water stress than V_Cmax.     

气孔导度对CO2浓度变化的模拟及其生理机制

基于气孔运动的生理生化机制重点进行了气孔导度(gs)对CO2浓度变化的响应机制分析,并推导得到气孔导度(gs)对CO2浓度变化响应模型,并以9种植物进行了模型验证。结果表明:随着CO2浓度的升高,气孔导度会逐渐降低,且下降的幅度会随着CO2浓度的升高而逐渐减弱。气孔导度对CO2浓度(Cs)变化的响应模型可以表达为gs=gmax/(1+Cs/Cs0),其中式中gmax是最大气孔导度和Cs0是实验常数。该模型较好地模拟了气孔导度随CO2浓度变化的规律,模型参数具有明确的生理意义,与Jarvis模型和Ball-Berry模型相比,该模型如何实现多种环境因子的耦合有待进一步突破。另外,模型是在短期改变叶片CO2浓度的条件下得出的,在CO2浓度长期胁迫下的适用性也有待进一步确认。     

5-Aminolaevulinate dehydratase of wheat leaves: Distribution and response to light

5-Aminolaevulinate dehydratase occurs almost exclusively in the stroma of etioplasts and chloroplasts of young wheat leaves and is absent from mitochondria or cytosol fractions. Etiolated plants show little or no change in activity of the enzyme during the first 30 hr of illumination except for a small transient increase in the first hour which may be under phytochrome control.     

On measuring the response of mesophyll conductance to carbon dioxide with the variable J method

The response of mesophyll conductance to CO(2) (g(m)) to environmental variation is a challenging parameter to measure with current methods. The 'variable J' technique, used in the majority of studies of g(m), assumes a one-to-one relationship between photosystem II (PSII) fluorescence and photosynthesis under non-photorespiratory conditions. When calibrating this relationship for Populus trichocarpa, it was found that calibration relationships produced using variation in light and CO(2) were not equivalent, and in all cases the relationships were non-linear-something not accounted for in previous studies. Detailed analyses were performed of whether different calibration procedures affect the observed g(m) response to CO(2). Past linear and assumed calibration methods resulted in systematic biases in the fluorescence estimates of electron transport. A sensitivity analysis on modelled data (where g(m) was held constant) demonstrated that biases in the estimation of electron transport as small as 2% (～0.5 μmol m(-2) s(-1)) resulted in apparent changes in the relationship of g(m) to CO(2) of similar shape and magnitude to those observed with past calibration techniques. This sensitivity to biases introduced during calibrations leads to results where g(m) artefactually decreases with CO(2), assuming that g(m) is constant; if g(m) responds to CO(2), then biases associated with past calibration methods would lead to overestimates of the slope of the relationship. Non-linear calibrations were evaluated; these removed the bias present in past calibrations, but the method remained sensitive to measurement errors. Thus measurement errors, calibration non-linearities leading to bias, and the sensitivity of variable J g(m) hinders its use under conditions of varying CO(2) or light.