Diversity in leaf anatomy, and stomatal distribution and conductance, between salt marsh and freshwater species in the C\u3csub\u3e4\u3c/sub\u3e genus Spartina (Poaceae)

Leaf anatomy, stomatal density, and leaf conductance were studied in 10 species of Spartina (Poaceae) from low versus high salt marsh, and freshwater habitats. • Internal structure, external morphology, cuticle structure, and stomatal densities were studied with light and electron microscopy. Functional significance of leaf structure was examined by measures of CO2 uptake and stomatal distributions. • All species have Kranz anatomy and C 4 δ13C values. Freshwater species have thin leaves with small ridges on adaxial sides and stomata on both adaxial and abaxial sides. By contrast, salt marsh species have thick leaves with very pronounced ridges on the adaxial side and stomata located almost exclusively on adaxial leaf surfaces. Salt marsh species also have a thicker cuticle on the abaxial than on the adaxial side of leaves, and CO2 uptake during photosynthesis is restricted to the adaxial leaf surface. • Salt marsh species are adapted to controlling water loss by having stomata in leaf furrows on the adaxial side, which increases the boundary layer, and by having large leaf ridges that fit together as the leaf rolls during water stress. Differences in structural-functional features of photosynthesis in Spartina species are suggested to be related to adaptations to saline environments

Knockdown of glycine decarboxylase complex alters photorespiratory carbon isotope fractionation in Oryza sativa leaves

The influence of reduced glycine decarboxylase complex (GDC) activity on leaf atmosphere CO2 and 13CO2 exchange was tested in transgenic Oryza sativa with the GDC H-subunit knocked down in leaf mesophyll cells. Leaf measurements on transgenic gdch knockdown and wild-type plants were carried out in the light under photorespiratory and low photorespiratory conditions (i.e. 18.4 kPa and 1.84 kPa atmospheric O2 partial pressure, respectively), and in the dark. Under approximately current ambient O2 partial pressure (18.4 kPa pO2), the gdch knockdown plants showed an expected photorespiratory-deficient phenotype, with lower leaf net CO2 assimilation rates (A) than the wild-type. Additionally, under these conditions, the gdch knockdown plants had greater leaf net discrimination against 13CO2 (Δo) than the wild-type. This difference in Δo was in part due to lower 13C photorespiratory fractionation (f) ascribed to alternative decarboxylation of photorespiratory intermediates. Furthermore, the leaf dark respiration rate (Rd) was enhanced and the 13CO2 composition of respired CO2 (δ13CRd) showed a tendency to be more depleted in the gdch knockdown plants. These changes in Rd and δ13CRd were due to the amount and carbon isotopic composition of substrates available for dark respiration. These results demonstrate that impairment of the photorespiratory pathway affects leaf 13CO2 exchange, particularly the 13C decarboxylation fractionation associated with photorespiration.Research was funded by a C4 Rice Project grant from The Bill and Melinda Gates Foundation to IRRI (2012–2015) and to the University of Oxford (2015–2019); by the National Science Foundation, grant MCB-1146928; by the National Science Foundation, grant MRI0923562; and by the Russian Science Foundation, grant 16-16-00089

Transgenic maize phosphoenolpyruvate carboxylase alters leaf-atmosphere CO2 and 13CO2 exchanges in Oryza sativa.

The engineering process of C4 photosynthesis into C3 plants requires an increased activity of phosphoenolpyruvate carboxylase (PEPC) in the cytosol of leaf mesophyll cells. The literature varies on the physiological effect of transgenic maize (Zea mays) PEPC (ZmPEPC) leaf expression in Oryza sativa (rice). Therefore, to address this issue, leaf-atmosphere CO2 and 13CO2 exchanges were measured, both in the light (at atmospheric O2 partial pressure of 1.84 kPa and at different CO2 levels) and in the dark, in transgenic rice expressing ZmPEPC and wild-type (WT) plants. The in vitro PEPC activity was 25 times higher in the PEPC overexpressing (PEPC-OE) plants (~20% of maize) compared to the negligible activity in WT. In the PEPC-OE plants, the estimated fraction of carboxylation by PEPC (β) was ~6% and leaf net biochemical discrimination against 13CO2[Formula: see text] was ~ 2‰ lower than in WT. However, there were no differences in leaf net CO2 assimilation rates (A) between genotypes, while the leaf dark respiration rates (Rd) over three hours after light-dark transition were enhanced (~ 30%) and with a higher 13C composition [Formula: see text] in the PEPC-OE plants compared to WT. These data indicate that ZmPEPC in the PEPC-OE rice plants contributes to leaf carbon metabolism in both the light and in the dark. However, there are some factors, potentially posttranslational regulation and PEP availability, which reduce ZmPEPC activity in vivo

Transgenic maize phosphoenolpyruvate carboxylase alters leaf-atmosphere CO2 and 13CO2 exchanges in Oryza sativa

The engineering process of C4 photosynthesis into C3 plants requires an increased activity of phosphoenolpyruvate carboxylase (PEPC) in the cytosol of leaf mesophyll cells. The literature varies on the physiological efect of transgenic maize (Zea mays) PEPC (ZmPEPC) leaf expression in Oryza sativa (rice). Therefore, to address this issue, leaf-atmosphere CO2 and 13CO2 exchanges were measured, both in the light (at atmospheric O2 partial pressure of 1.84 kPa and at diferent CO2 levels) and in the dark, in transgenic rice expressing ZmPEPC and wild-type (WT) plants. The in vitro PEPC activity was 25 times higher in the PEPC overexpressing (PEPC-OE) plants (~20% of maize) compared to the negligible activity in WT. In the PEPC-OE plants, the estimated fraction of carboxylation by PEPC (β) was ~6% and leaf net biochemical discrimination against 13CO2 (Δbio) was ~2‰ lower than in WT. However, there were no diferences in leaf net CO2 assimilation rates (A) between genotypes, while the leaf dark respiration rates (Rd) over three hours after light-dark transition were enhanced (~ 30%) and with a higher 13C composition (훿13CRd) in the PEPC-OE plants compared to WT. These data indicate that ZmPEPC in the PEPC-OE rice plants contributes to leaf carbon metabolism in both the light and in the dark. However, there are some factors, potentially posttranslational regulation and PEP availability, which reduce ZmPEPC activity in vivo.This research was funded by the C4 Rice Project grant from The Bill and Melinda Gates Foundation to IRRI (2012– 2015) and to the University of Oxford (2015–2019); by the National Science Foundation, Grant MCB-1146928; by the National Science Foundation, Grant MRI-0923562; and by the Russian Science Foundation, Grant 16-16-00089

Single cell C 4 photosynthesis in aquatic and terrestrial plants: A gas exchange perspective

Another "green revolution" is needed for crop yields to meet the demands for food and this has provided a new focus on photosynthesis research. Many important crops have the C3 photosynthetic pathway and a range of options for enhancing leaf photosynthesis in C3 species are being tested. George Bowes' pioneering research elucidated the inducible single cell C4 photosynthetic mechanism of aquatic plants such as Hydrilla verticillata. Here we review the efficacy of single cell C4 photosynthesis by comparing the physiology of terrestrial Kranz C4 photosynthesis with that of terrestrial and aquatic single cell C4 photosynthesis. The comparison shows that the terrestrial single cell system which involves spatially separated cytoplasmic domains and dimorphic chloroplasts equals Kranz C4 photosynthesis in efficiency; whereas, the simpler aquatic single cell system employed by H. verticillata which operates the C4 pathway between cytoplasmic space and chloroplasts is less efficient due to diffusive constraints. Nevertheless, understanding this readily inducible C4 pathway may provide valuable lessons for improving C3 photosynthesis by enhancing chloroplast CO2 concentrations

Biochemical and Structural Diversification of C₄ Photosynthesis in Tribe Zoysieae (Poaceae)

C4 photosynthesis has evolved independently multiple times in grass lineages with nine anatomical and three biochemical subtypes. Chloridoideae represents one of the separate events and contains species of two biochemical subtypes, NAD-ME and PEP-CK. Assessment of C4 photosynthesis diversification is limited by species sampling. In this study, the biochemical subtypes together with anatomical leaf traits were analyzed in 19 species to reveal the evolutionary scenario for diversification of C4 photosynthesis in tribe Zoysieae (Chloridoideae). The effect of habitat on anatomical and biochemical diversification was also evaluated. The results for the 19 species studied indicate that 11 species have only NAD-ME as a decarboxylating enzyme, while eight species belong to the PEP-CK subtype. Leaf anatomy corresponds to the biochemical subtype. Analysis of Zoysieae phylogeny indicates multiple switches between PEP-CK and NAD-ME photosynthetic subtypes, with PEP-CK most likely as the ancestral subtype, and with multiple independent PEP-CK decarboxylase losses and its secondary acquisition. A strong correlation was detected between C4 biochemical subtypes studied and habitat annual precipitation wherein NAD-ME species are confined to drier habitats, while PEP-CK species prefer humid areas. Structural adaptations to arid climate include increases in leaf thickness and interveinal distance. Our analysis suggests that multiple loss of PEP-CK decarboxylase could have been driven by climate aridization followed by continued adaptive changes in leaf anatomy