The literature and our present examinations indicate that the intra-leaf light absorption profile is in most cases steeper than the photosynthetic capacity profile. In strong white light, therefore, the quantum yield of photosynthesis would be lower in the upper chloroplasts, located near the illuminated surface, than that in the lower chloroplasts. Because green light can penetrate further into the leaf than red or blue light, in strong white light, any additional green light absorbed by the lower chloroplasts would increase leaf photosynthesis to a greater extent than would additional red or blue light. Based on the assessment of effects of the additional monochromatic light on leaf photosynthesis, we developed the differential quantum yield method that quantifies efficiency of any monochromatic light in white light. Application of this method to sunflower leaves clearly showed that, in moderate to strong white light, green light drove photosynthesis more effectively than red light. The green leaf should have a considerable volume of chloroplasts to accommodate the inefficient carboxylation enzyme, Rubisco, and deliver appropriate light to all the chloroplasts. By using chlorophylls that absorb green light weakly, modifying mesophyll structure and adjusting the Rubisco/chlorophyll ratio, the leaf appears to satisfy two somewhat conflicting requirements: to increase the absorptance of photosynthetically active radiation, and to drive photosynthesis efficiently in all the chloroplasts. We also discuss some serious problems that are caused by neglecting these intra-leaf profiles when estimating whole leaf electron transport rates and assessing photoinhibition by fluorescence techniques.

Rubisco is a large enzyme with a molecular mass of approximately 550 kD. The maximum rate of CO2 fixation (i.e. ribulose-1,5-bisphosphate [RuBP] carboxylation)

CO₂ faces a series of resistances while diffusing between the substomatal cavities and the sites of carboxylation within chloroplasts. The absence of techniques to measure the resistance of individual steps makes it difficult to define their relative importance. Resistance to diffusion through intercellular airspace differs between leaves, but is usually of minor importance. Leaves with high photosynthetic capacity per unit leaf area reduce mesophyll resistance by increasing the surface area of chloroplasts exposed to intercellular airspace per unit leaf area, S_c. Cell walls impose a significant resistance. Assuming an effective porosity of the cell wall of 0.1 or 0.05, then cell walls could account for 25% or 50% of the total mesophyll resistance, respectively. Since the fraction of apoplastic water that is unbound and available for unhindered CO₂ diffusion is unknown, it is possible that the effective porosity is 50% of the total resistance and a variable proportion. Most of the remaining resistance is imposed by one or more of the three membranes as mesophyll resistance can be altered by varying the expression of cooporins. The CO₂ permeability of vesicles prepared from chloroplast envelopes has been reduced by RNA interference (RNAi) expression of NtAQP1, but not those prepared from the plasma membrane. Carbonic anhydrase activity also influences mesophyll resistance. Mesophyll resistance is relatively insensitive to the manipulation of any step in the pathway because it represents only part of the total and may also be countered by pleiotropic compensatory changes. The parameters in greatest need of additional measurements are S_c, mesophyll cell wall thickness, and the permeabilities of the plasma membrane and chloroplast envelope.

The subject of this paper, sun leaves are thicker and show higher photosynthetic rates than the shade leaves, is approached in two ways. The first seeks to answer the question: why are sun leaves thicker than shade leaves? To do this, CO₂ diffusion within a leaf is examined first. Because affinity of Rubisco for CO₂ is low, the carboxylation of ribulose 1,5-bisphosphate is competitively inhibited by O₂, and the oxygenation of ribulose 1,5-bisphosphate leads to energy-consuming photorespiration, it is essential for C₃ plants to maintain the CO₂ concentration in the chloroplast as high as possible. Since the internal conductance for CO₂ diffusion from the intercellular space to the chloroplast stroma is finite and relatively small, C₃ leaves should have sufficient mesophyll surfaces occupied by chloroplasts to secure the area for CO₂ dissolution and transport. This explains why sun leaves are thicker. The second approach is mechanistic or ‘how-oriented’. Mechanisms are discussed as to how sun leaves become thicker than shade leaves, in particular, the long-distance signal transduction from mature leaves to leaf primordia inducing the periclinal division of the palisade tissue cells. To increase the mesophyll surface area, the leaf can either be thicker or have smaller cells. Issues of cell size are discussed to understand plasticity in leaf thickness.

The internal conductance for CO₂ diffusion (g_i) and CO₂ assimilation rate were measured and the related anatomical characteristics were investigated in transgenic rice leaves that overexpressed barley aquaporin HvPIP2;1. This study was performed to test the hypothesis that aquaporin facilitates CO₂ diffusion within leaves. The g_i value was estimated for intact leaves by concurrent measurements of gas exchange and carbon isotope ratio. The leaves of the transgenic rice plants that expressed the highest levels of Aq-anti-HvPIP2;1 showed a 40% increase in g_i as compared to g_i in the leaves of wild-type rice plants. The increase in g_i was accompanied by a 14% increase in CO₂ assimilation rate and a 27% increase in stomatal conductance (g_s). The transgenic plants that had low levels of Aq-anti-HvPIP2;1 showed decreases in g_i and CO₂ assimilation rate. In the plants with high levels of Aq-anti-HvPIP2;1, mesophyll cell size decreased and the cell walls of the epidermis and mesophyll cells thickened, indicating that the leaves had become xeromorphic. Although such anatomical changes could partially offset the increase in g_i by the aquaporin, the increase in aquaporin content overcame such adverse effects.