Fotosintesis
Fotosintesis
Fotosintesis
Glossary
LHCII The light-harvesting complex of photosystem II, PsbS A protein that is involved in the NPQ mechanism of
a protein–chlorophyll complex within the thylakoid heat dissipation.
membrane that captures light energy and transfers it to the RuBisCO The enzyme ribulose-1,5-bisphosphate (RuBP)
photosystem reaction centers. carboxylase/oxygenase, which carboxylates RuBP to form
NPQ Nonphotochemical quenching of chlorophyll a three-carbon sugar. In C3 plants, RuBisCO also catalyzes
fluorescence due to dissipation of excess chlorophyll the oxygenation of RuBP, the initial step in
excitation energy as heat. This process competes with photorespiration.
productive photochemical processes and photosystem II
fluorescence.
The continuing rise in atmospheric CO2 concentrations is Oxygenic photosynthesis is the sunlight-driven process used by
driving a rapid increase in ambient temperatures. The accompa- photoautotrophic organisms to assimilate atmospheric carbon
nying environmental changes will progressively reduce the area dioxide (CO2) into organic molecules. In land plants, the
of arable land available and thus pose a grave threat to global photosynthetic reactions occur within chloroplasts, organelles
food security. The situation is exacerbated both by exponential that are derived from an endosymbiotic event that was initiated
population growth and increased demand for crop plants as when a cyanobacterial ancestor was engulfed by a eukaryotic
sources of renewable energy or high-value products. The foresee- host cell. Photosynthesis can be divided into two major sets
able intensification of competition between agronomical and of reactions – a strictly light-driven electron transport process
industrial use makes it imperative that the available supply of associated with the thylakoid membranes which generates
cropland be used more efficiently. During the Green Revolution ATP and NADPH (light reactions), and the Calvin–Benson
that began in the 1960s, significant increases in yield could be (CB) cycle by which CO2 is incorporated into carbohydrates
achieved by more effective farming strategies, innovations in in the chloroplast stroma. From an electrochemical point of
fertilization, and the introduction of dwarfing genes into impor- view, H2O serves as the ultimate electron donor and CO2 as
tant crop species like rice (Oryza sativa) and wheat (Triticum aes- the final electron acceptor in this redox reaction.
tivum). The last resulted in a shift of carbon allocation within the
plant from the vegetative tissue to the grain. The stunted growth
Electron Transfer across the Thylakoid Membrane
phenotypes of the new varieties also reduced yield losses caused
(Light Reactions)
by fertilization-based lodging effects. In recent years, conven-
tional breeding endeavors have been unable to maintain the The electron transport reactions are driven by the two major
resulting high rates of grain yield increase per unit area of arable pigment-containing multiprotein complexes, photosystem II
land, and the beneficial effects associated with the Green Revolu- (PSII) and photosystem I (PSI), which are embedded in the thyla-
tion have virtually ceased due to the already widespread cultiva- koid membrane. The two systems differ in their light absorption
tion of improved varieties. properties, since they exhibit distinct excitation maxima in the
Currently the most promising approaches to improving crop low-red (PSII/680 nm) and far-red (PSI/700 nm) regions of the
yield appear to be those based on the genetic engineering of visible spectrum (Figure 1). The corresponding accessory
developmental or bioenergetic processes, such as photosyn- light-harvesting complexes (LHCs), LHCII and LHCI, are made
thesis. These approaches offer the prospect of a renewal of the up of different proteins, chlorophylls (Chl a and Chl b, respec-
Green Revolution, which is urgently required to meet the contin- tively), carotenoids, and associated lipids.
uously increasing demand for superior high-yield crop varieties Linear electron flow (LEF) from PSII to PSI involves two
for human sustenance and industrial applications in the future. further complexes – the cytochrome b6f complex (Cyt b6f) and
This article will highlight genetic approaches to the remod- the ATP synthase – and is initiated by splitting of water and the
eling of the primary metabolism of photosynthesis with a view concomitant extraction of electrons at the oxygen-evolving
to establishing the basis for a sustainable increase in yield and complex (OEC) on the donor side of PSII. The electrons are trans-
biomass production in crop plants. ferred via the plastoquinone (PQ) pool to Cyt b6f and
Figure 1 Scheme depicting the course of the photosynthetic light reactions and the Calvin–Benson cycle. The processes of photosynthetic electron
transport and proton translocation (gray arrows) around the photosystems (PSII and PSI) are shown in the case of linear electron flow (LEF, red
arrows) and cyclic electron flow (CEF, blue arrows) which eventually lead to the generation of NADPH and ATP via the ATP synthase. The products of
the light reactions are preferentially consumed by the Calvin–Benson cycle (CB, brown arrows) which assimilates CO2 to form triose phosphates
(starch) from RuBP. The wasteful oxygenation reaction of RuBisCO (photorespiration) effectively causes a net loss in CO2 fixation. Abbreviations: Cyt
b6f, cytochrome b6f complex; Fd, ferredoxin; FNR, ferredoxin-NADPH oxidoreductase; PGRL1, PGRL1/PGR5 complex; PG, phosphoglycolate; P680/
P700, P680/P700 reaction center; PSI, photosystem I; PSII, photosystem II; PC, plastocyanin; PQ, plastoquinone; TM, thylakoid membrane.
subsequently passed on to plastocyanin (PC) and PSI. Further PSI and PSII. In this way, it is possible to equalize excitation of
steps in LEF comprise electron transfers from ferredoxin (Fd) to both photosystems, which is a requirement for optimal electron
a Fd-NADP reductase (FNR), which reduces NADPþ to NADPH transport across the thylakoid membrane (Figure 2(b)). The
and Hþ. LEF effectively results in the generation of a proton diverse regulatory mechanisms required to compensate for fluctu-
gradient, which powers the production of ATP by the ations in environmental conditions offer a range of starting points
membrane-bound enzyme ATP synthase, and ATP together for modifications aimed at streamlining photosynthesis in crop
with NADPH drives CO2 assimilation by the CB (Figure 1). The species grown under controlled light conditions.
assimilation of CO2 via the CB requires an ATP to NADPH ratio
of 3:2. However, LEF alone is insufficient to provide the necessary
CO2 Fixation (CB) and Photorespiration
ATP. Cyclic electron flow (CEF), in which electrons are shuttled
back from the acceptor side of PSI to the PQ pool – either via Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is
PGRL1/PGR5 or the NDH complex – thereby transferring protons the key enzyme involved in photosynthetic carbon fixation, as it
from the stroma to the lumen of the thylakoid membrane is catalyzes the conversion of atmospheric CO2 into organic
assumed to compensate for this shortfall (Figure 1). compounds. In the first step two molecules of
As sessile organisms, land plants are exposed to significant 3-phosphoglycerate (3-PGA) are produced, one of which is
fluctuations in ambient light conditions. To compensate for these required for the regeneration of ribulose-1,5-bisphosphate
variations, they have developed several regulatory mechanisms (RuBP), the substrate to which the enzyme attaches the CO2.
that enable them either to optimize the allocation of radiant In every third reaction cycle one molecule of 3-PGA is diverted
energy to the photosynthetic light reactions under light-limiting to sustain the biosynthesis of sugars and other high-energy
conditions or to protect their pigment–protein complexes from compounds (Figure 1). Under CO2 limitation and saturating
damage by high-intensity light. Excess energy absorbed by the light conditions, the CO2 fixation step by RuBisCO is regarded
photosynthetic complexes can, for instance, be dissipated as as the rate-limiting step in the photosynthetic process. Moreover,
heat via energy-dependent quenching (qE) mechanisms. One RuBisCO displays a significant affinity for molecular oxygen and
such mechanism involves the PsbS protein, protonation of which also catalyzes an oxygenation reaction that further diminishes
alters the conformation of the LHCII antennae associated with the overall efficiency of photosynthesis. At current atmospheric
PSII and induces a heat dissipative state. Another component CO2 concentrations and temperatures (25 C), about one-third
that contributes to qE is the enzyme violaxanthin de-epoxidase of the carbohydrate formed during C3 photosynthesis is
(VDE), which catalyzes the conversion of violaxanthin into the consumed by this photorespiration reaction. In this energetically
quenching-competent xanthophyll zeaxanthin. Both nonphoto- wasteful process RuBisCO reacts with molecular oxygen to form
chemical quenching (NPQ) mechanisms allow plants to respond one molecule of 3-PGA and one of 2-phosphoglycolate (PG).
to rapid fluctuations in light intensity (Figure 2(a)). In contrast, The latter acts as an inhibitor of CB enzymes and must be con-
under light-limiting growth conditions a mechanism referred to verted back into 3-PGA in an energy-intensive conversion reac-
as ‘state transitions’ allows plants to utilize the incident light tion (photorespiration) that requires shuttling between three
most efficiently by redistributing a certain fraction of their additional cell compartments/organelles – the cytosol, peroxi-
light-harvesting pigment–protein complexes (LHCs) between somes, and mitochondria – and produces CO2. Furthermore,
92 Photosynthesis j Photosynthesis
Figure 2 Adaptation of photosynthetic light reactions to the level of incident light. (a) Under high-light conditions, excess absorbed light energy can
be dissipated as heat rather quickly (qE). One mechanism that contributes to this process involves protonation of the PsbS protein, which in turn
alters the conformation and attachment of the antenna proteins associated with PSII (LHCII). A second protective mechanism is the conversion of
violaxanthin to zeaxanthin by the enzyme violaxanthin de-epoxidase. (b) Under light-limiting conditions the state transition mechanism (qT) allows for
equalization of excitation of the two photosystems by reallocating a mobile fraction of LHCII between PSII and PSI. This occurs in a reversible, LHCII
phosphorylation-dependent manner. Nonphosphorylated LHCII remains preferentially associated with PSII (State 1), while phosphorylated LHCII has
a higher affinity for PSI (State 2). Abbreviations: P, phosphate; TM, thylakoid membrane; VDE, violaxanthin de-epoxidase; ZE, zeaxanthin epoxidase.
the extent of loss of fixed CO2 gradually increases with a rise in which differ in their mode of CO2 fixation (Figure 3). Fixation
ambient temperature. of CO2 into the stable three-carbon compound 3-PGA, cata-
lyzed by RuBisCO, is essential for all plant species and is
responsible for the designation of C3 plants. The C3-type of
Variants of CO2 Fixation in Land Plants: C3, C4, photosynthesis (Figure 3(a)) is the most widespread among
and CAM Photosynthesis land plants and is found in various crop species such as wheat
(T. aestivum), rice (O. sativa), potatoes (Solanum tuberosum), and
In land plants, three photosynthetic pathways, termed C3, C4, sugar beet (Beta vulgaris). In C3 photosynthesis, a minimum of
and Crassulacean Acid Metabolism (CAM), are known to exist, three ATP and two NADPH molecules are required to
Figure 3 Principles of C3, C4, and Crassulacean Acid Metabolism (CAM) photosynthesis. (a) During C3 photosynthesis RuBisCO’s capacity for
oxygenation leads to formation of varying amounts of phosphoglycolate which undergoes photorespiration. (b) During C4 photosynthesis CO2 is first
fixed in the mesophyll cells as malate, which is then transferred to bundle sheath cells. There it is converted to pyruvate, effectively raising CO2
concentrations in the immediate vicinity of RuBisCO. (c) CAM photosynthesis allows for fixation of CO2 during the night, and the resulting malate is
subsequently stored in the vacuole (lilac). During the day malate is released from the vacuole and converted back to phosphoenolpyruvate, thereby
releasing CO2 for the carboxylation reaction of RuBisCO. Abbreviations: CA, carbonic anhydrase; M, malate; MDH, malate dehydrogenase; ME, malic
enzyme; OA, oxaloacetate; PEP, phosphoenolpyruvate; PEPC, phosphoenolpyruvate carboxylase; PG, phosphoglycolate; PR, photorespiration; Pyr,
pyruvate; PPDK, pyruvate phosphate dikinase.
Photosynthesis j Photosynthesis 93
assimilate one molecule of CO2 into carbohydrate and regen- In cyanobacteria, a slightly different type of organelle-like
erate one RuBP to complete the C3 cycle. Based on the blue- microbody harbors the enzymatic machinery for carbon fixa-
print of C3 photosynthesis, C4 and CAM plants have tion and establishes a CO2-enriched environment to suppress
developed mechanisms to reduce photorespiration. the wasteful oxygenation reaction of RuBisCO. These
In C4 plants, this goal is mainly achieved by using another so-called carboxysomes are surrounded by a proteinaceous
enzyme for initial fixation of CO2. Furthermore, the O2-gener- shell that encapsulates RuBisCO and carbonic anhydrases,
ating light reactions are spatially separated from CO2 fixation thereby providing an advantage in low-CO2 environments.
by RuBisCO, ensuring that the latter enzyme is exposed to The assembly of carboxysomes is well understood. In the first
lower levels of molecular oxygen. In all C4 plants the initial step, its core components – RuBisCO and CAs – form macro-
CO2 fixation step is mediated by phosphoenolpyruvate (PEP) molecular aggregates called procarboxysomes, which are then
carboxylase (PEPC), which catalyzes the fixation of bicarbonate coated with shell proteins at distinct sites to form functional
into oxaloacetate (OA). This is then converted into four carbon carboxysomes, and are distributed throughout the cell. The
acids like malate or aspartate, e.g., in the chloroplasts of meso- high efficiency of carboxysome-mediated carbon fixation
phyll cells (MC) (Figure 3(b)). These metabolites are then strongly depends on the biochemical properties of the shell
shuttled to the chloroplasts of bundle sheath cells (BSCs) structure, specifically on its permeability to the substrate of
and decarboxylated, generating a microenvironment in which RuBisCO (RuBP) and its product phosphoglycerate (PGA).
RuBisCO ‘sees’ elevated CO2 levels. Such CO2-concentrating Favorable influx kinetics of bicarbonate and CO2 further
mechanisms (CCMs) come at the expense of an extra two promote the carboxylation reaction of RuBisCO.
ATPs, are innate to C4 and CAM species, and evolved
several times among land plant species. The leaves of most
C4 plants – among them major crop species like maize (Zea Potential Targets for Enhancement of Photosynthesis
mays), sorghum (Sorghum bicolor), and sugarcane (Saccharum
Optimization/Streamlining of the Light Reactions
officinarum) – have developed a specialized morphology, the
so-called ‘Kranz’ anatomy: the vascular bundles are surrounded To improve photosynthetic performance in land plants, earlier
by two layers composed of different cell types. The inner layer studies mainly focused on the modification of single or small
consists of BSCs that contain chloroplasts with increased sets of genes involved in biosynthetic or regulatory pathways
amounts of RuBisCO and lack grana stacks due to a strong that had been identified as bottlenecks in processes related to
reduction in the level of PSII complexes. In contrast, the chlo- carbon assimilation. More recent approaches aim to streamline
roplasts in the MCs of the outer layer resemble those of C3 the photosynthetic light reactions and their associated regula-
plants. CAM photosynthesis occurs in only a few crops (e.g., tory mechanisms. One idea is simply to eliminate regulatory
pineapple (Ananas comosus)) and relies on the PEP processes – e.g., state transitions that allow plants adapt to vari-
carboxylase-dependent fixation of CO2 in the cytoplasm of able light conditions – that are dispensable when plants are
MCs but, unlike C4 species, CAM plants store the resulting grown under controlled greenhouse conditions (Figure 2(b)).
organic acids in vacuoles for later use (Figure 3(c)). CAM In principle, deleting all but the absolutely essential regulatory
plants fix and store CO2 in this way at night when respiration mechanisms should make more energy available for biomass
is the dominant reaction, and make it available for fixation in production and increasing yield. The beneficial effects on
the chloroplasts during the day. Thus they concentrate CO2 at photosynthesis, however, that might be generated by manipu-
certain times, while C4 plants concentrate CO2 spatially in lating dynamic regulatory processes are difficult to predict.
specialized cell types. Both C4 and CAM plants use water Therefore, a more direct approach would be to increase the
more efficiently than C3 plants. intrinsic efficiency of the components immediately involved
in the photosynthetic process, such as the multiprotein
complexes in the thylakoid membrane.
Carbon-Concentrating Mechanisms in Green Algae To achieve improvements in the efficacy of complex and
and Cyanobacteria evolutionarily conserved structures like the photosynthetic
complexes in crop plants, multiple genetic modifications,
In comparison to land plants, algae use different combinations rather than single gene alterations, will be required. However,
of CCMs in a species-dependent manner in order to increase approaches based on randomized variation of multiple genetic
CO2 concentrations in the vicinity of their carbon fixation loci are generally difficult to realize in crop species, due to their
machinery. Such mechanisms utilize energy-dependent trans- usually very long life cycle. To overcome the limitation of long
porters of inorganic carbon, carbonic anhydrases (CAs) to generation times, current approaches that seek to optimize the
release CO2 from bicarbonate, and in the case of various light reactions by modulating plant photosynthetic complexes
eukaryotic algae, spatial confinement of RuBisCO to pyrenoids per se are performed in cyanobacteria. Their short generation
– crystal-like proteinaceous structures – within the plastid. The times, and evolutionary conservation with respect to the
unifying feature of the morphologically variable pyrenoids is biochemical properties of land plant chloroplasts, render cya-
that RuBisCO is their dominant constituent, although other nobacteria eminently suitable for large-scale mutational
protein components like RuBisCO activase, nitrate reductase screens. A further advantage of cyanobacteria over land plants
(NR), fructose-1,6-bisphosphate aldolase (FBA), or CAs have as hosts for modifying plant-specific protein complexes or
been shown to be associated with pyrenoids in different enzymatic reactions lies in the fact that various types of genetic
species, suggesting a certain diversity in its biochemical manipulations are readily feasible. Features like homologous
properties. recombination, marker-free gene replacement, and the ability
94 Photosynthesis j Photosynthesis