Mesophyll conductance to CO2: current knowledge and future prospects

During photosynthesis, CO2 moves from the atmosphere (C(a)) surrounding the leaf to the sub-stomatal internal cavities (C(i)) through stomata, and from there to the site of carboxylation inside the chloroplast stroma (C(c)) through the leaf mesophyll. The latter CO2 diffusion component is called mesophyll conductance (g(m)), and can be divided in at least three components, that is, conductance through intercellular air spaces (g(ias)), through cell wall (g(w)) and through the liquid phase inside cells (g(liq)). A large body of evidence has accumulated in the past two decades indicating that g(m) is sufficiently small as to significantly decrease C(c) relative to C(i), therefore limiting photosynthesis. Moreover, g(m) is not constant, and it changes among species and in response to environmental factors. In addition, there is now evidence that g(liq) and, in some cases, g(w), are the main determinants of g(m). Mesophyll conductance is very dynamic, changing in response to environmental variables as rapid or even faster than stomatal conductance (i.e. within seconds to minutes). A revision of current knowledge on g(m) is presented. Firstly, a historical perspective is given, highlighting the founding works and methods, followed by a re-examination of the range of variation of g(m) among plant species and functional groups, and a revision of the responses of g(m) to different external (biotic and abiotic) and internal (developmental, structural and metabolic) factors. The possible physiological bases for g(m), including aquaporins and carbonic anhydrases, are discussed. Possible ecological implications for variable g(m) are indicated, and the errors induced by neglecting g(m) when interpreting photosynthesis and carbon isotope discrimination models are highlighted. Finally, a series of research priorities for the near future are proposed.     

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.     

Complex adjustments of photosynthetic potentials and internal diffusion conductance to current and previous light availabilities and leaf age in Mediterranean evergreen species Quercus ilex

Mature non-senescent leaves of evergreen species become gradually shaded as new foliage develops and canopy expands, but the interactive effects of integrated light during leaf formation (Q(int)G), current light (Q(int)C) and leaf age on foliage photosynthetic competence are poorly understood. In Quercus ilex L., we measured the responses of leaf structural and physiological variables to Q(int)C and Q(int)G for four leaf age classes. Leaf aging resulted in increases in leaf dry mass per unit area (M(A)), and leaf dry to fresh mass ratio (D(F)) and decreases in N content per dry mass (N(M)). N content per area (N(A)) was independent of age, indicating that decreases in N(M) reflected dilution of leaf N because of accumulation of dry mass (NA = N(M) M(A)). M(A), D(F) and N(A) scaled positively with irradiance, whereas these age-specific correlations were stronger with leaf growth light than with current leaf light. Area-based maximum ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylase activity (V(cmax)A), capacity for photosynthetic electron transport (J(max)A) and the rate of non-photorespiratory respiration in light (R(d)A) were also positively associated with irradiance. Differently from leaf structural characteristics, for all data pooled, these relationships were stronger with current light with little differences among leaves of different age. Acclimation to current leaf light environment was achieved by light-dependent partitioning of N in rate-limiting proteins. Mass-based physiological activities decreased with increasing leaf age, reflecting dilution of leaf N and a larger fraction of non-photosynthetic N in older leaves. This resulted in age-dependent modification of leaf photosynthetic potentials versus N relationships. Internal diffusion conductance (g(m)) per unit area (g(m)A) increased curvilinearly with increasing irradiance for two youngest leaf age classes and was independent of light for older leaves. In contrast, g(m) per dry mass (g(m)M) was negatively associated with light in current-year leaves. Greater photosynthetic potentials and moderate changes in diffusion conductance resulted in greater internal diffusion limitations of photosynthesis in higher light. Both area- and mass-based g(m) decreased with increasing leaf age. The decrease in diffusion conductance was larger than changes in photosynthetic potentials, leading to larger CO2 drawdown from leaf internal air space to chloroplasts (delta(c)) in older leaves. The increases in diffusion limitations in older leaves and at higher light scaled with age- and light-dependent increases in MA and D(F). Overall, our study demonstrates a large potential of foliage photosynthetic acclimation to changes in leaf light environment, but also highlights enhanced structural diffusion limitations in older leaves that result from leaf structural acclimation to previous rather than to current light environment and accumulation of structural compounds with leaf age.     

Temperature response of mesophyll conductance in cultivated and wild Oryza species with contrasting mesophyll cell wall thickness

A critical component of photosynthetic capacity is the conductance of CO(2) from intercellular airspaces to the sites of CO(2) fixation in the stroma of chloroplasts, termed mesophyll conductance (g(m)). Leaf anatomy has been identified as an important determinant of g(m). There are few studies of the temperature response of g(m) and none has examined the implications of leaf anatomy. Hence, we compared a cultivar of Oryza sativa with two wild Oryza relatives endemic to the hot northern savannah of Australia, namely Oryza meridionalis and Oryza australiensis. All three species had similar leaf anatomical properties, except that the wild relatives had significantly thicker mesophyll cell walls than O. sativa. Thicker mesophyll cell walls in the wild rice species are likely to have contributed to the reduction in g(m) , which was associated with a greater drawdown of CO(2) into chloroplasts (C(i) -C(c) ) compared with O. sativa. Mesophyll conductance increased at higher temperatures, whereas the rate of CO(2) assimilation was relatively stable between 20 and 40 °C. Consequently, C(i) -C(c) decreased for all three species as temperature increased.     

The influence of leaf thickness on the CO2 transfer conductance and leaf stable carbon isotope ratio for some evergreen tree species in Japanese warm-temperate forests

1. The influence of leaf thickness on internal conductance for CO₂ transfer from substomatal cavity to chloroplast stroma ( g _i) and carbon isotope ratio (δ¹³C) of leaf dry matter was investigated for some evergreen tree species from Japanese temperate forests. g _i was estimated based on the combined measurements of gas exchange and concurrent carbon isotope discrimination.
2. Leaves with thicker mesophyll tended to have larger leaf dry mass per area (LMA), larger surface area of mesophyll cells exposed to intercellular air spaces per unit leaf area ( S _mes) and smaller volume ratio of intercellular spaces to the whole mesophyll (mesophyll porosity).
3. g _i of these leaves was correlated positively to S _mes but negatively to mesophyll porosity. The variation in g _i among these species would be therefore primarily determined by variation of the conductance in liquid phase rather than that in gas phase.
4. δ¹³C was positively correlated to mesophyll thickness and leaf nitrogen content on an area basis. However, g _i values did not correlate to δ¹³C. These results suggest that difference in δ¹³C among the species was not caused by the variation in g _i, but mainly by the difference in long-term photosynthetic capacity.
5. Comparison of our results with those of previous studies showed that the correlation between leaf thickness and g _i differed depending on leaf functional types (evergreen, deciduous or annual). Differences in leaf properties among these functional types were discussed.     

Anatomical basis of variation in mesophyll resistance in eastern Australian sclerophylls: news of a long and winding path

In sclerophylls, photosynthesis is particularly strongly limited by mesophyll diffusion resistance from substomatal cavities to chloroplasts (r (m)), but the controls on diffusion limits by integral leaf variables such as leaf thickness, density, and dry mass per unit area and by the individual steps along the diffusion pathway are imperfectly understood. To gain insight into the determinants of r (m) in leaves with varying structure, the full CO(2) physical diffusion pathway was analysed in 32 Australian species sampled from sites contrasting in soil nutrients and rainfall, and having leaf structures from mesophytic to strongly sclerophyllous. r (m) was estimated based on combined measurements of gas exchange and chlorophyll fluorescence. In addition, r (m) was modelled on the basis of detailed anatomical measurements to separate the importance of different serial resistances affecting CO(2) diffusion into chloroplasts. The strongest sources of variation in r (m) were S (c)/S, the exposed surface area of chloroplasts per unit leaf area, and mesophyll cell wall thickness, t (cw). The strong correlation of r (m) with t (cw) could not be explained by cell wall thickness alone, and most likely arose from a further effect of cell wall porosity. The CO(2) drawdown from intercellular spaces to chloroplasts was positively correlated with t (cw), suggesting enhanced diffusional limitations in leaves with thicker cell walls. Leaf thickness and density were poorly correlated with S (c)/S, indicating that widely varying combinations of leaf anatomical traits occur at given values of leaf integrated traits, and suggesting that detailed anatomical studies are needed to predict r (m) for any given species.     

Leaf structure and specific leaf mass: the alpine desert plants of the Eastern Pamirs, Tadjikistan

This study examines interrelationships between eight leaf attributes (specific leaf mass, area, dry mass, lamina thickness, mesophyll cell number per cm², mesophyll cell volume, chloroplast volume, and number of chloroplasts per mesophyll cell) in field-grown plants of 94 species from the Eastern Pamir Mountains, at elevations between 3800 and 4750 m. Unlike most other mountain areas, the Eastern Pamirs, Karakorum system, Tadjikistan provide localities where low temperatures and radiation combine with moisture stress at high altitudes. For all the attributes measured, significant differences were found between plants with different mesophyll types. Leaves with dorsiventral palisade structure (dorsal palisade, ventral spongy mesophyll cells) had thicker leaves with larger but fewer mesophyll cells, containing more and larger chloroplasts. These differences in mesophyll type are reflected in differences in the total surface of mesophyll cells per unit leaf area ( A _mes/ A ) or volume ( A _mes/ V ). Plants with isopalisade leaf structure (palisade cells under both dorsal and ventral surfaces) are more commonly xerophytes and their increased values of A _mes/ A and A _mes/ V decrease CO₂ mesophyll resistance, which is an important adaptation to drought. Path analysis shows the critical importance of mesophyll cell volume in leading to the covariance between the different leaf attributes and hence to specific leaf mass (SLM), even though mesophyll cell volume is not itself strongly correlated with SLM. This is because mesophyll cell volume increases SLM through its effects on leaf thickness and chloroplast number per cell, but decreases SLM through its negative effect on mesophyll cell density.     

On the relationship between leaf anatomy and CO2 diffusion through the mesophyll of hypostomatous leaves

Internal conductances to CO₂ transfer from the stomatal cavity to sites of carboxylation (g_i) in hypostomatous sun-and shade-grown leaves of citrus, peach and Macadamia trees (Lloyd et al. 1992) were related to anatomical characteristics of mesophyll tissues. There was a consistent relationship between absorptance of photosynthetically active radiation and chlorophyll concentration (mmol m^?2) for all leaves, including sclerophyllous Macadamia, whose transmittance was high despite its relatively thick leaves. In thin peach leaves, which had high g_i, the chloro-plast volume and mesophyll surface area exposed to intercellular air spaces (ias) per unit leaf area were similar to those in the thicker leaves of the evergreen species. Peach leaves, however, had the lowest leaf dry weight per area (D/a), the lowest tissue density (T_d) and the highest chloro-plast surface area (S_c) exposed to ias. There were negative correlations between g_i and leaf thickness or D/a, but positive correlations between g_i and S_c or S_c/T_d. We developed a one-dimensional diffusion model which partitioned g_i into a gaseous diffusion conductance through the ias (g_ias) plus a liquid-phase conductance through mesophyll cell walls (g_cw). The model accounted for a significant amount of variation (r₂=0.80) in measured g_i by incorporating both components. The g_ias component was related to the one-dimensional path-length for diffusion across the mesophyll and so was greater in thinner peach leaves than in leaves of evergreen species. The g_cw component was related to tissue density and to the degree of chloroplast exposure to the ias. Thus the negative correlations between g_i and leaf thickness or D/a related to g_ias whereas positive correlations between g_i and S_c or S_c/T_d, related to g_cw. The g_cw was consistently lower than g_ias, and thus represented a greater constraint on CO₂ diffusion in the mesophylls of these hypostomatous species.     

Leaf anatomy as a constraint for photosynthetic acclimation: differential responses in leaf anatomy to increasing growth irradiance among three deciduous trees

Interspecific variation in the response to transfer from low to high growth irradiance with respect to anatomical and photosynthetic characteristics was studied in mature leaves of three tree species, Betula ermanii Cham., Acer rufinerve Sieb. et Zucc. and Fagus crenata Blume, which occur in different successional stages in temperate deciduous forests. Transfer from low to high irradiance increased the light-saturated rate of photosynthesis per unit leaf area ( P _max) significantly in B. ermanii and A. rufinerve , but not in F. crenata . Leaves of B. ermanii grown at low irradiance were relatively thick and had vacant spaces along the mesophyll cell surfaces which was not occupied by chloroplasts or other organelles. After transfer to high irradiance, chloroplasts enlarged to fill the space along with P _max without an increase in leaf thickness. Leaves of A. rufinerve were plastic in mesophyll cell surface area and in leaf thickness, both of which increased after the transfer to high irradiance, along with an increase in the amount of chloroplasts and in P _max. On the other hand, F. crenata had little mesophyll cell surface unoccupied by chloroplasts and leaf anatomy was not changed after the transfer. In all species, P _max was strongly correlated with chloroplast surface area adjacent to the exposed mesophyll surface across different growth irradiances. An increase in P _max was observed only when chloroplast volume also increased. We conclude that light acclimation potential is primarily determined by the availability of unoccupied cell surface into which chloroplasts expand, as well as by the plasticity of the mesophyll that allows an increase in its surface area.     

Structural assessment of the impact of environmental constraints on Arabidopsis thaliana leaf growth: a 3D approach

Light and soil water content affect leaf surface area expansion through modifications in epidermal cell numbers and area, while effects on leaf thickness and mesophyll cell volumes are far less documented. Here, three-dimensional imaging was applied in a study of Arabidopsis thaliana leaf growth to determine leaf thickness and the cellular organization of mesophyll tissues under moderate soil water deficit and two cumulative light conditions. In contrast to surface area, thickness was highly conserved in response to water deficit under both low and high cumulative light regimes. Unlike epidermal and palisade mesophyll tissues, no reductions in cell number were observed in the spongy mesophyll; cells had rather changed in volume and shape. Furthermore, leaf features of a selection of genotypes affected in leaf functioning were analysed. The low-starch mutant pgm had very thick leaves because of unusually large palisade mesophyll cells, together with high levels of photosynthesis and stomatal conductance. By means of an open stomata mutant and a 9-cis-epoxycarotenoid dioxygenase overexpressor, it was shown that stomatal conductance does not necessarily have a major impact on leaf dimensions and cellular organization, pointing to additional mechanisms for the control of CO(2) diffusion under high and low stomatal conductance, respectively.     

Shading-induced changes in the leaf mesophyll of plants of different functional types

Changes in the structural characteristics of mesophyll induced by shading were investigated in ten species of wild plants of diverse functional types. In all plant types, shading reduced leaf thickness and density by 30–50% and total surface of mesophyll, by 30–70%. The extent and mechanisms of mesophyll structural rearrangement depended on the plant functional type. In the ruderal plants, integral parameters of mesophyll, such as the surface of cells and chloroplasts and mesophyll resistance, changed threefold predominantly because of changes in the dimensions of the cells and chloroplasts. In these plants, shading reduced the volume of chloroplasts by 30%, and the chloroplast numbers per cell declined. The competitor plants showed a twofold increase in mesophyll resistance due to a decrease in the number of photosynthesizing cells per leaf area unit. Moreover, these plants maintained constant dimensions of mesophyll cells, ratios mesophyll surface/mesophyll volume and chloroplast surface/cell surface. In stress-tolerant plants, diffusion resistance of mesophyll remained the same irrespective of the growing conditions, and mesophyll rearrangement was associated with inversely proportional changes in the dimensions of the cells and cell volume per chloroplast. Noteworthy of these plants were relatively constant chloroplasts number per cell, per leaf area unit and total surface area of chloroplasts. The nature of relationship between the mesophyll diffusion resistance and structural parameters of leaf mesophyll differed in plants of diverse functional types.     

Tobacco aquaporin NtAQP1 is involved in mesophyll conductance to CO2 in vivo

Leaf mesophyll conductance to CO(2) (g(m)) has been recognized to be finite and variable, rapidly adapting to environmental conditions. The physiological basis for fast changes in g(m) is poorly understood, but current reports suggest the involvement of protein-facilitated CO(2) diffusion across cell membranes. A good candidate for this could be the Nicotiana tabacum L. aquaporin NtAQP1, which was shown to increase membrane permeability to CO(2) in Xenopus oocytes. The objective of the present work was to evaluate its effect on the in vivo mesophyll conductance to CO(2), using plants either deficient in or overexpressing NtAQP1. Antisense plants deficient in NtAQP1 (AS) and NtAQP1 overexpressing tobacco plants (O) were compared with their respective wild-type (WT) genotypes (CAS and CO). Plants grown under optimum conditions showed different photosynthetic rates at saturating light, with a decrease of 13% in AS and an increase of 20% in O, compared with their respective controls. CO(2) response curves of photosynthesis also showed significant differences among genotypes. However, in vitro analysis demonstrated that these differences could not be attributed to alterations in Rubisco activity or ribulose-1,5-bisphosphate content. Analyses of chlorophyll fluorescence and on-line (13)C discrimination indicated that the observed differences in net photosynthesis (A(N)) among genotypes were due to different leaf mesophyll conductances to CO(2), which was estimated to be 30% lower in AS and 20% higher in O compared with their respective WT. These results provide evidence for the in vivo involvement of aquaporin NtAQP1 in mesophyll conductance to CO(2).     

Estimating near-infrared leaf reflectance from leaf structural characteristics

The relationship between near-infrared reflectance at 800 nm (NIRR) from leaves and characteristics of leaf structure known to affect photosynthesis was investigated in 48 species of alpine angiosperms. This wavelength was selected to discriminate the effects of leaf structure vs. chemical or water content on leaf reflectance. A quantitative model was first constructed correlating NIRR with leaf structural characteristics for six species, and then validated using all 48 species. Among the structural characteristics tested in the reflectance model were leaf trichome density, the presence or absence of both leaf bicoloration and a thick leaf cuticle (>1 μm), leaf thickness, the ratio of palisade mesophyll to spongy mesophyll thickness (PM/SM), the proportion of the mesophyll occupied by intercellular air spaces (%IAS), and the ratio of mesophyll cell surface area exposed to IAS (A(mes)) per unit leaf surface area (A), or A(mes)/A. Multiple regression analysis showed that measured NIRR was highly correlated with A(mes)/A, leaf bicoloration, and the presence of a thick leaf cuticle (r = 0.93). In contrast, correlations between NIRR and leaf trichome density, leaf thickness, the PM/SM ratio, or %IAS were relatively weak (r < 0.25). A model incorporating A(mes)/A, leaf bicoloration, and cuticle thickness predicted NIRR accurately for 48 species (r = 0.43; P < 0.01) and may be useful for linking remotely sensed data to plant structure and function.     

Development of leaf thickness for Plectranthus parviflorus – Influence of photosynthetically active radiation

Photosynthetically active radiation (PhAR) is apparently the environmental factor having the greatest influence on leaf thickness for Plectranthus parviflorus Henckel (Labiatae). A four-fold increase in leaf thickness from 280 to 1170 μm occurred as the PhAR was raised from 1.3 to 32.5 mol m⁻² day⁻¹. Compared to a constant PhAR of 2.5 mol m⁻² day⁻¹, a PhAR of 32.5 mol m⁻² day⁻¹ for one week during the first week (with return to 2.5 mol m⁻² day⁻¹ during the second and third weeks) led to an increase in final leaf thickness by 323 μm (to 802 μm). When increased PhAR was applied during the second week the increase in final thickness over the control was 217 μm, and when increased PhAR was applied during the third week it was 99 μm. However, leaf thickness was not simply responding to total daily PhAR, since a leaf 450 μm thick could occur at a low instantaneous PhAR for a long daytime (total daily PhAR of 1.5 mol m⁻² day⁻¹) and at a high PhAR for a short daytime (4.5 mol m⁻² day⁻¹). Total daily CO₂ uptake (net photosynthesis) was approximately the same in the two cases, suggesting that this is an important factor underlying the differences in leaf thickness. Leaf thickness is physiologically important, since thicker leaves tend to have greater mesophyll surface area per unit leaf area ( A ^mes/ A ) and hence higher photosynthetic rates.     

岷江上游干旱河谷海拔梯度上白刺花叶片生态解剖特征研究

对岷江上游干旱河谷海拔梯度上（1 650～1 950 m）白刺花(Sophora davidii)叶片进行生态解剖学研究.观测指标包括叶片形态特征（叶长宽比、叶面积、叶片厚度）、解剖结构（表皮厚度、栅栏组织厚度(P)、海绵组织厚度(S)、P/S比值、表皮角质膜厚度）及叶表皮特征（气孔器密度和面积、表皮细胞密度和面积、表皮毛密度和长度）.结果表明,白刺花叶片面积为0.144～0.208 cm²,叶总厚度为171.58～195.83 μm;叶肉组织分化明显,栅栏组织厚度与海绵组织厚度分别为69.83～82.42和62.00～ 80.67 μm,P/S的比值为1.14～1.01,上下表皮厚度分别为14.03～15.33和13.88～16.17 μm,上下角质膜厚度分别为2.66～4.56和2.76～2.02 μm;气孔密度为13.71～15.02个·mm^-2,其面积为249.86～280.43 μm²;表皮细胞密度为160.54～178.43个·mm^-2,其面积为557.43～626.85 μm²;表皮毛长度为186.51～260.99 μm,其密度为18.29～32.27个·mm^-2.随海拔升高叶面积、叶厚度、栅栏组织和海绵组织的厚度、气孔器面积、表皮细胞面积以及表皮毛密度呈增加趋势,而角质膜厚度、表皮细胞密度和表皮毛长度则呈减小趋势;叶长宽比、P/S的比值、表皮厚度与气孔器密度无明显差异.     

Temperature dependence of leaf-level CO2 fixation: revising biochemical coefficients through analysis of leaf three-dimensional structure

CO2 fixation in a leaf is determined by biochemical and physical processes within the boundaries set by leaf structure. Traditionally determined temperature dependencies of biochemical processes include physical processes related to CO2 exchange that result in inaccurate estimates of parameter values. A realistic three-dimensional model of a birch (Betula pendula) leaf was used to distinguish between the physical and biochemical processes affecting the temperature dependence of CO2 exchange, to determine new chloroplastic temperature dependencies for V c(max) and Jmax based on experiments, and to analyse mesophyll diffusion in detail. The constraint created by dissolution of CO2 at cell surfaces substantially decreased the CO2 flux and its concentration inside chloroplasts, especially at high temperatures. Consequently, newly determined chloroplastic V c(max) and Jmax were more temperature dependent than originally. The role of carbonic anhydrase in mesophyll diffusion appeared to be minor under representative mid-day nonwater-limited conditions. Leaf structure and physical processes significantly affect the apparent temperature dependence of CO2 exchange, especially at optimal high temperatures when the photosynthetic sink is strong. The influence of three-dimensional leaf structure on the light environment inside a leaf is marked and affects the local choice between Jmax and V c(max)-limited assimilation rates.     

Slow development of leaf photosynthesis in an evergreen broad-leaved tree, Castanopsis sieboldii: relationships between leaf anatomical characteristics and photosynthetic rate

Changes in net photosynthetic rate on a leaf area basis and anatomical properties during leaf development were studied in an evergreen broad‐leaved tree, Castanopsis sieboldii and an annual herb, Phaseolus vulgaris. In C. sieboldii, surface area of mesophyll cells facing the intercellular air spaces on a leaf area basis (S_mes) was already considerable at the time of full leaf area expansion (FLE). However, surface area of chloroplasts facing the intercellular air spaces on a leaf area basis (S_c), and chlorophyll and Rubisco contents on a leaf area basis increased to attain their maximal values 15–40 d after FLE. In contrast, in P. vulgaris, chloroplast number on a leaf area basis, S_c and S_mes at 10 d before FLE were two to three times greater than the steady‐state levels attained at around FLE. In C. sieboldii, the internal CO₂ transfer conductance (g_i) slightly increased for 10 d after FLE but then decreased toward the later stages. Limitation of photosynthesis by g_i was only about 10% at FLE, but then increased to about 30% at around 40 d after FLE. The large limitation after FLE by g_i was probably due to the decrease in CO₂ concentration in the chloroplast caused by the increases in thickness of mesophyll cell walls and in Rubisco content per chloroplast surface area. These results clearly showed that: (1) in C. sieboldii, chloroplast development proceeded more slowly than mesophyll cell expansion and continued well after FLE, whereas in P. vulgaris these processes proceeded synchronously and were completed by FLE; (2) after FLE, photosynthesis in leaves of C. sieboldii was markedly limited by g_i. From these results, it is suggested that, in the evergreen broad‐leaved trees, mechanical protection of mesophyll cells has priority over the efficient CO₂ transfer and quick construction of the chloroplasts.     

Mesophyll conductance to CO2 in Arabidopsis thaliana

The close rosette growth form, short petioles and small leaves of Arabidopsis thaliana make measurements with commercial gas exchange cuvettes difficult. This difficulty can be overcome by growing A. thaliana plants in 'ice-cream cone-like' soil pots. This design permitted simultaneous gas exchange and chlorophyll fluorescence measurements from which the first estimates of mesophyll conductance to CO(2) (g(m)) in Arabidopsis were obtained and used to determine photosynthetic limitations during plant ageing from c. 30-45 d. Estimations of g(m) showed maximum values of 0.2 mol CO(2) m(-2) s(-1) bar(-1), lower than expected for a thin-leaved annual species. The parameterization of the response of net photosynthesis (A(N)) to chloroplast CO(2) concentrations (C(c)) yielded estimations of the maximum velocity of carboxylation (V(c,max_Cc)) which were also lower than those reported for other annual species. As A. thaliana plants aged from 30 to 45 d, there was a 40% decline of A(N) that was entirely the result of increased diffusional limitations to CO(2) transfer, with g(m) being the largest. The results suggest that in A. thaliana A(N) is limited by low g(m) and low capacity for carboxylation. Decreased g(m) is the main factor involved in early age-induced photosynthetic decline.     

Mesophyll conductance and its limiting factors in plant leaves

Mesophyll conductance (g_m) represents the CO₂ diffusion facility from sub-stomatal internal cavities to carboxylation sites in chloroplasts, and the variation of g_m across genotypes as well as environmental conditions is expected to be related to the anatomical structures and biochemical properties of leaves. In recent years, the variation of g_m has attracted wide attention. The limiting factors in photosynthetic rate are no longer divided simply into stomatal limitation and non-stomatal limitation, but splitted in stomatal limitation, mesophyll limitation and carboxylation limitation. In this review, we summarize the potential influences of cell wall, cell membrane, cytoplasm, chloroplast envelope and stroma on g_m, and indicate that cell wall thickness and the surface area of chloroplast exposed to intercellular air space (S_c) are the most important factors influencing the g_m. We also analyze the probable effects of biochemical process related with aquaporins and carbonic anhydrase on g_m. Meanwhile, the regulation mechanisms of long- and short-term environment changes (including temperature, light intensity, drought, and nutrients) on g_m are also summarized. The relationship between g_m and hydraulic conductance (K_leaf) is debated. Finally, we discuss the scientific problems related with g_m.     

Effects of irradiance and spectral quality on leaf structure and function in seedlings of two Southeast Asian Hopea (Dipterocarpaceae) species

We studied the development of leaf characters in two Southeast Asian dipterocarp forest trees under different photosynthetic photon flux densities (PFD) and spectral qualities (red to far-red, R:FR). The two species, Hopea helferi and H. odorata, are taxonomically closely related but differ in their ecological requirements; H. helferi is more drought tolerant and H. odorata more shade tolerant. Seedlings were grown in replicated shadehouse treatments of differing PFD and R:FR. We measured or calculated (1) leaf and tissue thicknesses; (2) mesophyll parenchyma, air space, and lignified tissue volumes; (3) mesophyll air volumes (V(mes)/A(surf)) and surfaces (A(mes)/A(surf)); (4) palisade cell length and width; (5) chlorophyll/cm and a/b; (6) leaf absorption; and (7) attenuance/absorbance at 652 and 550 nm. These characters varied in response to light conditions in both taxa. Characters were predominantly affected by PFD, and R:FR slightly influenced many characters. Leaf characters of H. odorata were more plastic in response to treatment conditions. Characters were correlated with each other in a complex fashion. Variation in leaf anatomy is most likely a consequence of increasing leaf thickness in both taxa, which may increase mechanical strength and defense against herbivory in more exposed environments. Variation in leaf optical properties was most likely affected by pigment photo-bleaching in treatments of more intense PFD and was not correlated with A(max). The greater plasticity of leaf responses in H. odorata helps explain the acclimation over the range of light conditions encountered by this shade-tolerant taxon. The dense layer of scales on the leaf undersurface and other anatomical characters in H. helferi reduced gas exchange and growth in this drought-tolerant tree.