Chapter 4
Chapter 4
Chapter 4
4.1. Leaves are photosynthetic machines which maximize the absorption of light
4.2. Photosynthesis is an oxidation-reduction process
4.3. Photosynthetic electron transport
4.4. Photophosphorylation is the light-dependent synthesis of ATP
4.5. Lateral heterogeneity is the unequal distribution of thylakoid complexes
4.6. Light-harvesting complexes are super antenna complexes that regulate energy
distribution
4.7. Photoinhibition of photosynthesis: photoprotection versus photodamage
4.8. Inhibitors of photosynthetic electron transport are effective herbicides
Purpose
The organization of leaves with respect to the exploitation of light as the
primary source of energy and its conversion to the stable, chemical forms to
ATP and NADPH by the chloroplast
Special topics
- The structure of higher plant leaves with respect to the interception of light
- Photosynthesis as the reduction of CO2 to carbohydrate
- The photosynthetic electron transport chain, its organization in the thylakoid
membrane, and its role in generating reducing potential and ATP
- Problems encountered by chloroplasts when they are subject to varying
amounts of light, often in excess, which may decrease the efficiency of
photosynthesis or even damage components of the electron transport chain
- The dynamic nature of the thylakoid membrane, showing how changes in
the organization of light-harvesting apparatus influence the absorption and
distribution of light energy
- The role of carotenoids are accessory pigments and in photoprotection of
chlorophyll
- The use of herbicides that specifically interact with photosynthetic electron
transport
4.1. LEAVES ARE PHOTOSYNTHETIC MACHINES
THAT MAXIMIZE THE ABSORPTION OF LIGHT
The architecture of a typical higher plant leaf is particular well suited to absorb light
Palisade cells generally have larger numbers of chloroplasts than spongy mesophyll cells.
Because the absorbing pigments are confined to the chloroplast, a substantial amount of
light may thus pass through the first cell layer without being absorbed. This is called sieve
effect
The impact of the sieve effect on the efficiency of light absorption is balanced by factors (1)
light may all be reflected off the many surfaces associated with leaf cells (2) light that is not
reflected but passed between the aqueous volume of mesophyll cells and the spaces that
surround them will be sent by refraction (3) light may be scattered
The longer light path increases the probability that any given photon will be absorbed by a
chlorophyll molecule before it can escape from the leaf
Some of the incident light is channeled through the intercellular spaces between the palisade
cells in much the same way that light is transmitted by an optical fiber (fig4.2)
Within the leaf mesophyll cells of plants, the chloroplast is the organelle that transforms
light energy into ATP and NADPH to convert CO2 and sugar
Fig 4.1: The structure of leaves shown in cross-section
Fig 4.2: A simplified diagram illustrating how the optical properties of leaves help to
redistribute incoming light and maximize interception by chlorophyll
(A) Photon strikes a chloroplast and is absorbed by chlorophyll. (B) The sieve effect – a
photon passes through the first layer of mesophyll cells without being absorbed. It may
be absorbed in the next layer of cells or pass through the leaf to be absorbed by another
leaf below. (C) The plano-convex nature of epidermal cells creates a lens effects,
redirecting incoming light to. (D) The light-guide effect. Because the refractive index of
cells is greater than that of air, light reflected at the cell-air interfaces may be channeled
through the palisade layer(s) to the spongy mesophyll below
4.2. PHOTOSYNTHESIS IS AN OXIDIZATION-REDUCTION PROCESS
Addition of energy for carbon reduction is required ATP, which is also generated at the
expense of light
The key to the photosynthetic electron transport chain is the presence of 2 large, multimolecular
complexes photosystem I (PSI) and photosystem II (PSII) (fig4.3) that operate in series linked by a
third multiprotein aggregate called the cytochrome complex
Such fraction studies have revealed that PSI and PSII, each contain several different proteins together
with a collection of chlorophyll and carotenoid molecule that absorb photons
The bulk of the chlorophyll in the photosystem functions as antenna chlorophyll (fig4.4)
The association of chlorophyll with specific proteins forms a number of different chlorophyll protein
(CP) complexes (fig4.5)
The energy of absorbed photons thus migrates through the antenna complex, passing from 1
chlorophyll molecule to another until it eventually arrives at the reaction center (fig4.4)
Each reaction center consists of a unique chlorophyll a molecule that is though to be present as a
dime.
This reaction center chlorophyll + protein + redox carriers = light-driven redox reactions
The principal adventage of associating a sinle reaction center + a large number of antenna
chlorophyll molecules is to increase efficiency in the collection and utilization of light energy
A schematic of the photosynthetic electron transport chain depicting the arrangement of:
PSII PSI cytochrome b6/f (in thylakoid membrane) (fig4.6)
Fig 4.3: A linear representation of the photosynthetic electron-transport chain
A sequential arrangement of the 3 multimolecular membrane complexes extracts low-
energy electrons from water and, using light energy, produces a strong reductant,
NADPH + H+
Fig 4.4: A photosystem contains atenna and a reaction center
Antenna chlorophyll molecules absorb incoming photons and transfer the excitation
energy to the reaction center where the photochemical oxidation-reduction reaction occur
Fig 4.5:
Separation of thylakoid
chlorophyll-protein
complexes by
nondenaturing
polyacrylamide gel
electrophoresis
Fig 4.5a: Separation of thylakoid chlorophyll-protein complexes
by nondenaturing polyacrylamide gel electrophoresis
(A) In the presence of specific detergents, chlorophyll-protein complexes are removed from
thylakoid membranes structurally and functionally intact. These pigment-protein-detergent
complexes are charged and thus will migrate when an electric field is applied. The porous
matrix through which the electric field is applied is a polyacrylamide gel. Thus, the physical
separation of protein complexes through a polyacrylamide gel matrix by applying an electric
fields is called polyacrylamide gel electrophoresis. Since these proteins complexes still have
chlorophyll bound to them, you can watch the chlorophyll-protein complexes separate through
the gel matrix according to their molecular mass right in front of your eyes ! The largest
complexes remain at the top of the gel and the smallest complexes near the bottom of the
gel. The illustration shows such an electrophoretic separation with the arrow indicating the
direction of migration. Typically, 7 “green bands” can be solved. Bands 1-6 are individual
chlorophyll-protein complexes associated with PSI and PSII. The band exhibiting the greatest
migration (band 7) is free pigment
Fig 4.5b: Separation of thylakoid chlorophyll-protein complexes
by nondenaturing polyacrylamide gel electrophoresis
(B) The relative amount represented by each green band can be qualified be scanning the
gel in a spectrophotometer. The peak areas provide an estimate of the relative content of
each chlorophyll-protein complex. Thus, green bands 2 and 6 are present at the greatest
amount in this particular thylakoid sample
Fig 4.5c: Separation of thylakoid chlorophyll-protein complexes
by nondenaturing polyacrylamide gel electrophoresis
(C) The pigment composition of each green band can be assessed by measuring the absorption
spectrum of each band. Here, the absorption spectrum of green bands 2, 5 and 6 only are illustrated.
Note that green bands 2 and 5 exhibit a similar absorption spectrum with maxima at about 435nm and
671nm. This indicates that both of these pigment-protein complexes have chlorophyll a bound to them.
Green band 2 represents PSI plus its core antenna and green band 5 represents the PSII core complex
(CP43 + CP47 + PSII reaction center). Green band 6 exhibits absorption maxima at 435nm and 671nm
representing chlorophyll a plus maxima 470nm and 654nm, representing chlorophyll b. Thus, green
band 6 contains both chlorophyll a and chlorophyll b. This green band represents LHCII, the major
chlorophyll a/b light-harvesting complex associated with PSII. Note that LHCII and PSDI are present in
the greatest amounts in this particular thylakoid sample
Fig 4.6: The organization of the photosynthetic electron transport system
in the thylakoid membrane
4.3.2. Photosystem II (PS II) oxidizes water to produce O2
Pheophytin is the primary electron acceptor in PSII. Photo-oxidation event result P680+
and Pheo-, a charge separation
The role of reaction proteins, D1 and D2, is to bind and to orient specific redox
carriers of the PSII reaction center in such a way as to decrease the probability of
charge recombination between P680+ and Pheo-
(1) pheophytin passes 1 electron on to a quinone (QA) forming [P680+ Pheo- QA] (fig4.7)
then the electron passed from QA plastoquinone (PQ) forming [P680+ Pheo QA]
(2) P680+ reduced P680 forming [P680 Pheo QA]
P680+ was supplied the electrons by a cluster of 4 Mg2+ ions associated with a small
complex protein called the oxygen-evolving complex (OEC). OEC is bound to the D1
and D2 proteins of PSII reaction center and functions to stabilize Mg2+ cluster
2H2O O2 + 4H+ + 4e (4.8)
Pheophytin (Pheo)
Following its release from PSII plastoquinone (PQ) diffuse electron cytochrome
b6/f complex (contain s an additional redox component called Rieske ion-sulfur (FeS)
protein) From the Cyt f, electrons are pick up by plastocyanins (PC)
The primary electron acceptor in PSI is chlorophyll a passed through quinone (Q)
FeS center ferredoxin reduce NADP+
The over all effect of the complete electron transport scheme is to establish a
continuous flow of electron between water and NADP+, passing through PSII and PSI
and intervening cytochrome complex (fig4.6)
Water PSII cytochrome complex PSI NADP+ (Fig4.8)
The ATP required for carbon reduction and other metabolic activities of the chloroplast is
synthesized by photophosphorylation in accordance with Mitchell’s chemiosmotic
mechanism.
PSI has a process of cyclic electron transport (fig4.9) called cyclic photophosphorylation:
4 protons are translocated from stroma lumen:
ferredoxin carry electron back through Fdx-PQ to PQ FeS / Cyt f PC
There is a distinct lateral heterogeneity with respect to their distribution of the major
protein complexes within the thylakoids (fig4.11)
Both urea and triazine (fig4.17) herbicides are taken up by the roots and
transported to the leaves