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Evolution of C4 photosynthesis in response to changes in carbon and oxygen concentrations in the atmosphere through time.

The hypothesis is presented that C4 photosynthesis has evolved at lease in part in response to reduced efficiency of C3 photosynthesis due to increased oxygen partial pressures in our recent atmosphere. Oxygen competition with carbon dioxide for the active site on ribulose diphosphate carboxylase was of little consequence when atmospheric levels of CO2 were high and those of O2 were low. However at the present low CO2 and high O2 partial pressures, adaptations for protecting the enzyme from oxygen have been selected.

Atmosphere

Inhibition of oxalacetate decarboxylation during C4 photosynthesis by 3-mercaptopicolin acid.

3-Mercaptopicolinic acid specifically inhibits phosphoenolpyruvate carboxykinase in leaves of the C4 plant Panicum maximum. Both the ATP- and ADP-dependent decarboxylation of oxalacetate and the carboxylation activity of phosphoenolpyruvate carboxykinase are inhibited by 3-mercaptopicolinic acid while phosphoenolpyruvate carboxylase and ribulose-1,5-bisphosphate carboxylase are not inhibited. 3-Mercaptopicolinic acid inhibits the fixation of 14CO2 by illuminated P. maximum bundle sheath strands which is dependent upon oxalacetate and ATP but does not affect C3 photosynthesis in bundle sheath strands nor C4 photosynthesis in mesophyll cells. 3-Mercaptopicolinic acid treatment reduced P. maximum leaf photosynthesis 25% while raising the photosynthetic CO2 compensation point from near zero to 18 to 45 mul of CO2/liter of air.

Kinetics

Bundle sheath cell-specific expression of chloroplast genes encoding subunits of the NADH dehydrogenase-like complex in maize.

C4 photosynthesis alleviates the limitation caused by the oxygenase activity of Rubisco by partitioning photosynthetic functions between two distinct cell types: bundle sheath cells (BSCs) and mesophyll cells (MCs). These cell types perform different steps of photosynthesis using specialized machinery, accompanied by differential expression of chloroplast genes. To uncover the underlying molecular mechanisms for this differentiation, we isolated BSCs and MCs and compared their chloroplast transcriptomes, focusing on the chloroplast NADH dehydrogenase-like (NDH) complex, which is enriched in BSCs. To investigate whether RNA stabilization contributes to differential gene expression, we analyzed RNA footprints that reflect the binding of pentatricopeptide repeat (PPR) proteins to their RNA targets. We could not detect cell-type-specific accumulation of footprint RNAs. We then focused on transcriptional regulation, specifically on an operon that starts with the rps15 gene. The operon includes six ndh genes and the psaC gene encoding a photosystem I subunit. Transcript levels of all genes in this operon were higher in BSCs than in MCs, suggesting coordinated regulation as a transcriptional unit. Based on the genomic location of the rps15 gene within inverted repeats near the junctions on both sides of the small single copy region, we demonstrated that rps15, through two distinct promoters, is sufficient to drive preferential accumulation of downstream transcripts in BSCs.

Zea mays

Unraveling the molecular choreography of C3 to CAM transition in Mesembryanthemum crystallinum using phosphoproteomics.

Climate change and population growth threaten global freshwater resources and food security. Crassulacean acid metabolism (CAM) is a specialized photosynthetic adaptation that exhibits superior water use efficiency (WUE) compared to C3 and C4 photosynthesis. Mesembryanthemum crystallinum (common ice plant) is capable of shifting from C3 to CAM, making it a key model for investigating photosynthesis plasticity and its potential to enhance crop stress resilience. To date, the molecular mechanisms underlying this high-WUE photosynthetic transition remain largely unknown. Using mass spectrometry-based proteomics and phosphoproteomics, we quantified 4233 phosphopeptides containing 4758 phosphorylation sites, including the well-characterized Serine 11 of phosphoenolpyruvate carboxylase 1 (PEPC1). It is a critical phosphorylation site facilitating nocturnal CO2 fixation during CAM. Our analysis revealed many phosphorylation sites that exhibited similar diel patterns as the PEPC1 pS11, and they may be part of the regulatory network involved in CAM induction. Glycolysis/gluconeogenesis and carbon storage/breakdown modules exhibited extensive phosphorylation regulation, and vesicle trafficking could play a role in nocturnal carbon fixation. Furthermore, glycine-rich RNA-binding protein 7 (GRP7) in association with cold shock protein 1 (CSP1) emerged as a potential transcriptional switch for nocturnal stomatal opening. On the other hand, ABI5-binding protein 1 (AFP1) and oxidative stress 3 (OXS3)-activated ABA signaling, along with high CO2 signaling and suppressed blue light signaling, may contribute to diurnal stomatal closure. These findings shed light on the protein phosphorylation changes and provide valuable targets for functional characterization of their roles in CAM induction.

Mesembryanthemum

Photosynthetic carbon metabolism in Panicum milioides, a C3-C4 intermediate species: evidence for a limited C4 dicarboxylic acid pathway of photosynthesis.

Panicum milioides, a naturally occurring species with C4-like Kranz leaf anatomy, is intermediate between C3 and C4 plants with respect to photo-respiration and the associated oxygen inhibition of photosynthesis. This paper presents direct evidence for a limited degree of C4 photosynthesis in this C3-C4 intermediate species based on: (a) the appearance of 24% of the total 14C fixed following 4 s photosynthesis in 14CO2-air by excised leaves in malate and aspartate and the complete transfer of label from the C4 acids to Calvin cycle intermediates within a 15 s chase in 12CO2-air; (b) pyruvate- or alanine-enhanced light-dependent CO2 fixation and pyruvate stimulation ote- or alanine-enhanced light-dependent CO2 fixation and pyruvate stimulation of oxaloacetate- or 3-phosphoglycerate-dependent O2 evolution by illuminated mesophyll protoplasts, but not bundle sheath strands; and (c) NAD-malic enzyme-dependent decarboxylation of C4 acids at the C-4 carboxyl position, C4 acid-dependent O2 evolution, and 14CO2 donation from (4-14C)C4 acids to Calvin cycle intermediates during photosynthesis by bundle sheath strands, but not mesophyll protoplasts. However, P. milloides differs from C4 plants in that the activity of the C4 cycle enzymes is only 15 to 30% of a C4 Panicum species and the Calvin cycle and phosphoenolpyruvate carboxylase are present in both cell types. From these and related studies (Rathnam, C.K.M. and Chollet, R. (1979) Arch. Biochem. Biophys. 193, 346-354; (1978) Biochem. Biophys. Res. Commun. 85, 801-808) we conclude that reduced photorespiration in P. milioides is due to a limited degree of NAD-malic enzyme-type C4 photosynthesis permitting an increase in pCO2 at the site of bundle sheath, but not mesophyll, ribulose-bisphosphate carboxylase-oxygenase.

Aspartic Acid

The relationship of CO2 assimilation pathways and photorespiration to the physiological quantum requirement of green plant photosynthesis.

The quantum requirement of green cells for CO2 fixation has been evaluated and discussed in view of the recent discovery of photorespiration and of multiple biochemical pathways for photosynthetic CO2 fixation. The reported quantum requirement of algae generally is near 9 quanta per CO2 fixed. It is suggested that the high CO2 concentrations and low O2 concentrations used for these algae experiments would have completely suppressed photorespiration and, therefore, the minimum number of quanta required to fix 1 CO2 molecule was correctly determined in these experiments. With higher plant leaves, when measurements are made under physiological environments, quantum requirements range from about 12 to 20 quanta per CO2 fixed. It is suggested that these physiological quantum requirements are higher because photorespiration is functional in these leaves and that photorespiration requires energy. The energy requirement of photorespiration was derived using biochemical models of leaf photosynthesis combining photorespiration with specific biochemical pathways for CO2 fixation. The calculated physiological quantum requirements for C3, C4 and CAM plant photosynthesis are 13, 15 and 17 respectively. The literature values on quantum requirements correspond well with these biochemical models of net photosynthesis. However, it was concluded that the biochemical models fail to give a complete description of photosynthesis in plants using the C4-dicarboxylic acid cycle.

Carbon Dioxide

Photosynthesis and increased production of protein.

Photosynthesis, the use of light energy in the conversion of CO2 and inorganic nutrients into plant material, is the ultimate source of the food protein necessary to man's existence. Given certain assumptions, the overall maximal theoretical photosynthetic efficiency of agricultural plants can be calculated. Actual measured maximal growth rates of plants are equivalent to efficiency levels well below that theoretical maximum. In air, C4 plants can some closer to the theoretical value than C3 plants, perhaps because C4 plants avoid the occurrence of measurable photorespiration and oxygen inhibition of photosynthesis. Alfalfa, a C3 legume, is an extremely productive protein source. Its protein yield per acre can surpass that of commonly grown C4 crops (corn, sorghum) and C3 seed crops (soybean, wheat, rice). Alfalfa leaf protein is of high nutritional quality and can apparently be used directly in the human diet, eliminating the protein loss involved in animal production. Plant protein productivity can be raised as part of an increase in overall crop yield. The growth of plants in atmospheres with elevated CO2 levels can result in increased yields. In C3 plants this is due, at least in part, to the suppression of photorespiration and oxygen inhibition of photosynthesis. We have investigated the effect of CO2 concentration on alfalfa photosynthetic metabolism. Our results support the contention that alfalfa productivity can be increased by an environment of elevated CO2. A second approach toward increased plant protein productivity is through regulation of carbon flow during photosynthesis so as to increase portein production relative to that of other plant constituents. In particular, we have investigated whether ammonia (the form in which plants first incorporate nitrogen) can act to regulate leaf carbon metabolism. Our results indicate that NH4+, in part through stimulation of pyruvate kinase, brings about increased production of amino acids at the expense of sucrose production in alfalfa. The effect may be of considerable importance in the regulation of green leaf protein synthesis.

Amino Acids

A highactivity ATP translocator in mesophyll chloroplasts of Digitaria sanguinalis, a plant having the C-4 dicarboxylic acid pathway of photosynthesis.

The effect of exogenous adenine nucleotides on CO2 fixation and oxygen evolution was studied with mesophyll protoplast extracts of the C4 plant Digitaria sanguinalis. Exogenous ATP was found to stimulate the rate of pyruvate and pyruvate+oxalacetate induced CO2 fixation, as well as reverse the inhibition of CO2 fixation of carbonyl cyanide m-chlorophenyl hydrazone and several electron transport inhibitors. The ATP-dependent stimulation of CO2 fixation varied from 40 to 70 mumol CO2 fixed/mg chlorophyll per h, suggesting that ATP was crossing the chloroplast membranes at rates of 80-140 mumol/mg chlorophyll per h, since 2 ATP are required for each CO2 fixed. Fixation of CO2 could also be induced in the dark by exogenous ATP, in which case ADP accumulated outside the chloroplasts...

Adenosine Diphosphate