The genus Oryza, which includes rice (Oryza sativa and Oryza glaberrima) and wild relatives, is a useful genus to study leaf properties in order to identify structural features that control CO 2 access to chloroplasts, photosynthesis, water use efficiency, and drought tolerance. Traits, 26 structural and 17 functional, associated with photosynthesis and transpiration were quantified on 24 accessions (representatives of 17 species and eight genomes). Hypotheses of associations within, and between, structure, photosynthesis, and transpiration were tested. Two main clusters of positively interrelated leaf traits were identified: in the first cluster were structural features, leaf thickness (Thick leaf ), mesophyll (M) cell surface area exposed to intercellular air space per unit of leaf surface area (S mes ), and M cell size; a second group included functional traits, net photosynthetic rate, transpiration rate, M conductance to CO 2 diffusion (g m ), stomatal conductance to gas diffusion (g s ), and the g m /g s ratio. While net photosynthetic rate was positively correlated with g m , neither was significantly linked with any individual structural traits. The results suggest that changes in g m depend on covariations of multiple leaf (S mes ) and M cell (including cell wall thickness) structural traits. There was an inverse relationship between Thick leaf and transpiration rate and a significant positive association between Thick leaf and leaf transpiration efficiency. Interestingly, high g m together with high g m /g s and a low S mes /g m ratio (M resistance to CO 2 diffusion per unit of cell surface area exposed to intercellular air space) appear to be ideal for supporting leaf photosynthesis while preserving water; in addition, thick M cell walls may be beneficial for plant drought tolerance.

Four-year-old apple (Malus x domestica Borkh.) trees cv. 'Braeburn' on M.26 rootstock were thinned at full bloom to establish six crop loads ranging from a heavy crop to a deflowered treatment. At harvest, mean yield per tree varied from 0 to 38 kg and mean fruit weight ranged from 225 g in the heaviest cropping treatment to 385 g in the lightest cropping treatment. Light cropping resulted in a significant advance in fruit maturity as indicated by background color, starch/iodine score and soluble solids. There were small differences in leaf photosynthetic rate among the treatments when shoot growth was active. However, in early January, coincident with cessation of shoot growth and maximum rate of accumulation of fruit weight, leaf assimilation rate was reduced by as much as 65% on the deflowered trees compared to the trees carrying the heaviest crop. Leaf assimilation rate showed a curvilinear response to crop load at this time, with little increase in leaf assimilation when crop load exceeded 12 fruit m(-2) leaf area.

Summary Introduction of a C4 photosynthetic mechanism into C3 crops offers an opportunity to improve photosynthetic efficiency, biomass and yield in addition to potentially improving nitrogen and water use efficiency. To create a two‐cell metabolic prototype for an NADP‐malic enzyme type C4 rice, we transformed Oryza sativa spp. japonica cultivar Kitaake with a single construct containing the coding regions of carbonic anhydrase, phosphoenolpyruvate (PEP) carboxylase, NADP‐malate dehydrogenase, pyruvate orthophosphate dikinase and NADP‐malic enzyme from Zea mays, driven by cell‐preferential promoters. Gene expression, protein accumulation and enzyme activity were confirmed for all five transgenes, and intercellular localization of proteins was analysed. 13CO2 labelling demonstrated a 10‐fold increase in flux though PEP carboxylase, exceeding the increase in measured in vitro enzyme activity, and estimated to be about 2% of the maize photosynthetic flux. Flux from malate via pyruvate to PEP remained low, commensurate with the low NADP‐malic enzyme activity observed in the transgenic lines. Physiological perturbations were minor and RNA sequencing revealed no substantive effects of transgene expression on other endogenous rice transcripts associated with photosynthesis. These results provide promise that, with enhanced levels of the C4 proteins introduced thus far, a functional C4 pathway is achievable in rice.

Two-year-old Fagus sylvatica L. saplings were planted under the cover of a Pinus sylvestris L. stand in the French Massif Central. The stand was differentially thinned to obtain a gradient of transmitted photosynthetically active radiation (PAR(t); 0-0.35). Eighteen Fagus saplings were sampled in this gradient, and their growth (basal stem diameter increment) was recorded over six years. Over the same period, morphological parameters (leaf area, number and arrangement in space) were monitored by 3D-digitization. Photosynthetic parameters were estimated with a portable gas-exchange analyzer. Photosynthesis was mainly related to light availability, whereas sapling morphology was mainly driven by sapling size. Annual stem diameter increment was related to the amount of light-intercepting foliage (silhouette to total leaf area ratio (STAR) x total sapling leaf area (LA)) and light availability above the saplings (PAR(t)). However, light-use efficiency, i.e., the slope of the relationship between STAR x LA x PAR(t) and stem diameter increment, decreased over time as a result of a relative decrease in the proportion of photosynthetic tissues to total sapling biomass.

Summary The engineering of C4 photosynthetic activity into the C3 plant rice has the potential to nearly double rice yields. To engineer a two‐cell photosynthetic system in rice, the rice bundle sheath (BS) must be rewired to enhance photosynthetic capacity. Here, we show that BS chloroplast biogenesis is enhanced when the transcriptional activator, Oryza sativa Cytokinin GATA transcription factor 1 (OsCGA1), is driven by a vascular specific promoter. Ectopic expression of OsCGA1 resulted in increased BS chloroplast planar area and increased expression of photosynthesis‐associated nuclear genes (PhANG), required for the biogenesis of photosynthetically active chloroplasts in BS cells of rice. A further refinement using a DNAse dead Cas9 (dCas9) activation module driven by the same cell‐type specific promoter, directed enhanced chloroplast development of the BS cells when gRNA sequences were delivered by the dCas9 module to the promoter of the endogenous OsCGA1 gene. Single gRNA expression was sufficient to mediate the transactivation of both the endogenous gene and a transgenic GUS reporter fused with OsCGA1 promoter. Our results illustrate the potential for tissue‐specific dCas9‐activation and the co‐regulation of genes needed for multistep engineering of C4 rice.