Photosynthesis in Higher Plants

Photosynthesis is a physico-chemical process by which green plants use light energy to drive the
synthesis of organic compounds. It is an enzyme regulated anabolic process.

Light
6CO2 + 12 H2O C6H12O6 + 6H2O + 6O2
 Photosynthesis is the basis of life on earth because it is the primary source of all food on
earth and it is responsible for release of O2 in the atmosphere.
 Chlorophyll, light and CO2 is required for photosynthesis. It occurs only in green part of
leaves and in presence of light.

Early Experiments:

 Joseph Priestley in 1770, on the basis of his experiments showed the essential role of air in
growth of green plants. A mouse kept in closed space could get suffocated and die but if a
mint plant is kept in bell jar neither candle will extinguish nor will the mouse die. He
concluded that foul air produced by animal is converted into pure air by plants. Priestley
discovered Oxygen gas in 1774.


 Julius Von Sachs in 1854 shows that green part in plants produces glucose which is stored
as starch. Starch is the first visible product of photosynthesis.

 T.W.Engelmann (1843-1909) used prism to split light into its components and then
illuminated Cladophora (an algae) placed in a suspension of aerobic bacteria. He found that
 bacteria accumulated in blue and red light of the split spectrum. He thus discovered the effect
of different wavelength of light on photosynthesis (action spectrum).

 Cornelius Van Neil (1897-1985) on the basis of studies with purple and green sulphur
bacteria showed that photosynthesis is a light dependent reaction in which hydrogen from an
oxidisable compound reduces CO2 to form sugar.

Light
2H2A + CO2 2A + CH2O + H2O

In green sulphur bacteria, when H2S, instead of H2O was used as hydrogen donor, no O2 was
evolved. He inferred that O2 evolved by green plants comes from H2O but not from CO2 as
thought earlier.

Site of Photosynthesis

Photoshthesis takes place in the green leaves of plants, particularly in the chloroplast of their
mesophyll cells.

 Chloroplasts are green plastids which function as the site of photosynthesis in eukaryotic
photoautotrophs. Inside the leaves, chloroplast is generally present in mesophyll cells along
their walls.
 Within the chloroplast there is a membranous system consisting of grana, the stroma lamellae
and the fluid stroma.

 The membrane system is responsible for synthesizing light energy for the synthesis of ATP
and NADPH. In stroma enzymatic reactions incorporate CO2 in plants leading to synthesis of
sugar.
 The reaction in which light energy is absorbed by thylakoid (grana) to synthesis ATP and
NADPH is called light reaction (Non-cyclic and cyclic photophosphorylation). The later
part of photosynthesis in which CO2 is reduced to sugar, light is not necessary and is
called dark reaction (C3 or Calvin cycle) that occurs in stroma.

Pigments involved in Photosynthesis – Chromatographic separation of leaf pigments are as

follows-
Maximum absorption by chlorophyll a occurs in blue and red regions having higher rate of
photosynthesis. So, chlorophyll a is the chief pigment.

 Other thylakoid pigments like chlorophyll b, xanthophyll and carotenoids are

called accessary pigments that absorb light and transfer energy to chlorophyll a and protect
them from photo-oxidation.

Photosynthetic Apparatus

Pigment Light

Essential Non-essential

Takes part in e- transfer Takes part in energy transfer.

Eg.- Chl a (Reaction centre) All types of chlorophylls, carotenoids

and phycobilins except chl-a

Chlorophyll
 Chlorophylls are specialized lipid molecules embedded in thylakoid membrane.
 They are the main pigment concerned with the harvesting or trapping of solar energy.
 Arnoff and Allen recognized 9 types of chlorophyll: Chl-a, Chl-b, Chl-c, Chl-d, Chl-e,
bacteriochlorophyll-a, bacteriochlorophyll-b, chlorobium chlorophyll-650 and chlorobium
chlorophyll-666.
 Out of these, the 2 important types of chlorophyll found in green plants are chlorophyll-a and
chlorophyll-b.
 Chlorophyll-a: Universal pigment found in all green plants (C55H72O5N4Mg).
 Chlorophyll-b: Found in green algae and higher plants (C55H70O6N4Mg).
 Chlorophyll-c: Found in diatoms, dinoflagellates, and brown algae (C55H32O5N4Mg).
 Chlorophyll-d: Found in red algae.
 Chlorophyll molecule is asymmetrical and consists of 2 parts:
i) Porphyrin head ii) Phytol tail
 Chlorophyll-a is the most abundant photosynthetic pigment.
 It is the only pigment found in all photosynthetic plants.
 Chlorophylls absorb light near both ends of visible spectrum i.e. blue and red light, and
transmit or reflect green light. Therefore, these appear to be green in colour.

Synthesis of Chlorophyll
 Chlorophylls are synthesized from the precursors protochlorophyll.
 Its synthesis starts from glycine and succinyl CoA.
Carotenoids (Water insoluble)
 Carotenoids show brown, red, yellow and orange colours and are specialized lipids which
absorb light strongly in the blue-violet range.
 Carotenoids are terpenes, member of isoprene units that are water insoluble pigments.
 These are called shield pigments as they protect chlorophyll from photo-oxidation by light of
high intensity and also from oxygen produced during photosynthesis.
 These absorb light and transfer it to chlorophyll for use in photosynthesis and therefore
function as light harvesting complex.
Carotenoids are of 2 types:
i) Carotenes (C40H56): orange in colour.
 Carotenes are hydrocarbons, most of them are tetraterpenes. e.g., lycopene, α, β, γ-
carotenes.
ii) Xanthophylls (C40H56O2) are very similar to carotenes but contain oxygen, e.g.,
violaxanthin, fucoxanthin, Zeaxanthin.
Significance of Carotenoids:
 Β-carotene is precursor of vitamin-A.
 Conversion of atomic oxygen to molecular oxygen.
Phycobilins
 They are proteinaceous pigments found in phycobilisomes.
 They are soluble in hot water and do not contain magnesium and tail.
 Light is not essential for their synthesis.
 These pigments are accessory pigments and also help in chromatic adaptation.
Phytochrome
One type of phycobillins found in higher plants is phytochromes which takes part in
photomorphosynthetic movements like germination of photoblastic seeds, stomatal movement
and flowering responses.

Quantasome
 Park and Biggins coined the term quantasome for a group of pigment molecules required for
carrying out a photochemical reaction.
 These are situated as small units on the membranes of thylakoids.
 Each quantasome consists of about 230 chlorophyll molecules, carotenoids, quinone
compounds, sulpholipids, phospholipids, proteins, etc.
Absorption and Action Spectra
 Different plant pigments absorb only certain wavelengths of light and these wavelengths are
not absorbed at the same rate.
 A curve obtained by plotting the amount of absorption of different wavelengths of light by a
particular pigment is called absorption spectrum.
 An action spectrum is a curve showing the effectiveness of different wavelengths of light in
stimulating the process being investigated.
 The effectiveness of different wavelengths of light on photosynthesis is measured and plotted
by quantum yield or amount of action.

Graph of absorption spectra of chlorophyll-a, chlorophyll-b and carotenes

Graphical representation of differential photosynthetic effectiveness of different

wavelengths
Hill Reaction: Hill working on Stellaria media demonstrated that the evolution of O2 occurs
from illuminated chloroplast in the presence of hydrogen or electron acceptors but in the absence
of CO2, it produces assimilatory power for use in CO2 assimilation.
 Hill reaction is now considered to be equivalent to light reaction.
 Hill oxidants are hydrogen acceptors.
 The common ones are ferricyanide, benzoquinone, dichlorophenol indophenol, NADP+
(natural H+ acceptor in photosynthesis).
Quantum requirement: It is the number of photons required to produce one molecule of O2
photons.
Quantum yield conversion: It is the amount of O2 yield per photon = 12.5% Quantum
conversion amount of energy spent in photo excitation.
Red Drop and Emerson Enhancement Effect
 On exclusive exposure of more than 680 nm visible light, there is decline in quantum yield as
compared to natural quantum yield called red drop.
 On exclusive exposure of less than 680 nm visible light, although quantum yield increases as
compared to previous exposure, yet it is less than natural quantum yield.
 On simultaneous exposure of > 680 nm and < 680 nm light obtained, quantum yield is more
than sum total of individual light exposure. It is called enhancement effect. It is due to
synergistic effect of two distinct photosynthetic lengths (PSs).

Pigment Systems
 It is a group 250-400 pigment molecules having chlorophyll-a, chlorophyll-b, carotenoids.
 One molecule of chlorophyll-a functions as a reaction center or trapping center and the other
acts as light-harvesting complex (LHC) or antenna molecules.
 These antenna molecules collect light energy and transfer this energy to reaction center
where primary photochemical act occurs i.e., absorption of a photon and release of an
electron.
Mechanism of Photosynthesis
Photosynthesis occurs in two phases:
Light reaction
Light reaction (photochemical phase) includes:
1. Light absorption
2. Water splitting
3. Oxygen release
4. Formation of high energy chemical intermediates (ATP and NADPH).
 The pigments are organized into two discrete LHC (light harvesting complex) within
photosystem I and photosystem II, these are named in a sequence of their discovery.
 LHC are made up of hundreds of pigments molecules containing all pigments except single
chlorophyll a molecule in each PS.
 Each photosystem has all the pigments except chlorophyll a and forms an LHS called
antennae.
 The pigments in photosystem I and photosystem II absorb the lights of different wavelength.
Single chlorophyll a molecule makes the reaction centre. In PS I reaction centre has highest
peak at 700 nm, hence called P700. And PS II reaction centre has highest peak at 680 nm,
so called P680.

The Electron Transport System

In PS II LHC receives the light of 680 nanometers that results in the excitation of electrons and
thus they jump out of their orbit and are received by an electron acceptor that then passes it to the
electron transport system. Electrons are not used up and are passed to pigments of PS II with
simultaneous excitation of electrons in PS I because of absorption of light of 700 nanometers
these electrons are passed to a molecule of energy NADP+ resulting in the formation of NADPH
+ H+.
 1) Non-cyclic Photophosphorylation

OR

 The process by which ATP is synthesized by cells (in mitochondria and chloroplasts) is
named as phosphorylation.
 When the two photosystems work in a series, first PS II and the PS I, non-cyclic
photophosphorylation occurs.
 The 2 photosystems are connected through an electron transport chain. Both ATP and
NADPH + H+ are synthesized by this kind of electron flow.
 Reaction center of photosystem II absorbs light of 680 nm in red region and causing electron
to become excited. These electrons are picked by an electron acceptor which passes to
electron transport system consisting of cytochromes. This movement is downhill.
 Electrons are passed down the electron transport chain and then to the pigment of PS I.
 Electron in the PSI also get excited due to light of wavelength 700 nm and are transferred to
another accepter molecule having a greater redox potential.
 When electron passes in downhill direction, energy is released. This is used to reduce the
ADP to ATP and NADP+ to NADPH. The whole scheme of transfer of electron is called Z-
scheme due to its shape.

Splitting of Water

 The electrons that were moved from PS II are replaced by the splitting of H2O. This
creates Oxygen that is a product of photosynthesis.

2H2O ⟶ 4 H+ + O2 + 4e-

 The electrons needed to replace those removed from PS I are provided by PS II.
 This splitting of H2O is associated with PS II.

2) Cyclic Photophosphorylation
 OR

 Cyclic Photophosphorylation is a process of photophosphorylation in which an electron

expelled by the excited photocentre is returned back to it after passing through a series of
electron carriers.
 ATP is formed when electrons pass Ferredoxin to PQ and from PQ to the Cytochrome system.
 It occurs under conditions of low light intensity, wavelength longer than 680 nm and
when carbon dioxide fixation is inhibited.
 It is performed by PS I only.
 Its photocentere P700 extrudes an electron after absorbing a photon of light. After losing
the electron, the photocenter becomes oxidized. The expelled electron passes through a
series of carriers including P700 chlorophyll molecule, plastoquinone (PQ), FeS complex,
ferredoxin (Fd), cyt b6f and plastocyanin before returning to photocenter.
 Cyclic photophosphorylation occurs in both aerobic and anaerobic conditions.
 It is a process of producing carbohydrates by green plants using carbon dioxide and water in the
presence of sunlight.
Difference between cyclic and non-cyclic photophosphorylation

Cyclic photophosphorylation Non-cyclic photophosphorylation

1. It is performed by photosystem I 1. It is performed by collaboration of

independently. both PS I and PS II.
2. An external source of electron is 2. The process requires an external
not required. electron donor.
3. It synthesizes only ATP. 3. It synthesizes ATP and NADH both.
4. It occurs only in stromal or 4. It occurs in the granal thylakoids
intergranal thylakoids. only.

Chemiosmotic Hypothesis of ATP Formation

 This hypothesis was proposed by Mitchell in 1961.

 ATP synthesis is linked to development of proton gradient across the membrane of thylakoid
and mitochondria.
 The ATP synthase enzyme is made up of two parts: the F0 is introduced into the thylakoid
layer and produces a transmembrane channel that allows protons to diffuse across the layer
quite easily. The other half, known as F1, is located on the stroma-facing side of the
thylakoid membrane's exterior surface.
 The breakdown of the gradient provides enough energy to trigger a conformational shift in
the ATP synthase's F1 moiety, causing the enzyme to synthesis multiple molecules of ATP.
A membrane, a proton pump, a proton angle, and ATP synthase are all required for
chemiosmosis.
 The process that causes development of proton gradient across the membrane is-
1. Splitting of water molecules occurs inside the thylakoid to produce hydrogen ion
or proton.
2. As electron passes through the photosystems, protons are transported across the
membrane because primary acceptor of electron is located towards the outer side
the membrane transfers its electrons not to an electron carrier but to an H carrier.
As a result, this molecule removes a proton from the stroma while transporting an
electron.
3. When this molecule passes on its electron to the electron carrier on the inner side
of the membrane, the proton is released into the inner side (lumen) of the
membrane.
4. The NADP reductase enzyme is located in the stroma side of membrane.
Electrons that come out from the acceptor of electron of PSI along with protons
are necessary for reduction of NADP+ to NADP + H+. These protons are also
removed from the stroma.
5. This causes decrease in no.of protons in stroma while accumulation of protons
occur in the lumen.
6. This creates proton gradient across the thylakoid’s membrane along with pH in
the lumen.
7. Gradient is broken down due to movement of proton across the membrane to the
stroma through trans-membrane channel of F0 of ATPase. One part of this
enzyme is embedded in membrane to form trans-membrane channel. The other
 portion is called F1 that protrudes on the outer surface of thylakoid membrane on
the side that faces stroma.
8. The breakdown of the gradient provides enough energy to cause a conformational
change in the F1 particle of the ATPase which makes the enzyme synthesise
several molecules of energy packed ATP.
9. ATP and NADPH produced due to movement of electron is used immediately to
fix CO2 to form sugar.

 The product of light reaction used to drive the process leading to synthesis of
sugar are called biosynthetic phase of photosynthesis.

Calvin Cycle/C3 cycle/Reductive Pentose Sugar Phosphate Pathway:

Malvin Calvin, Benson and their colleagues used radioactive 14C and Chlorealla and
Scenedesmus algae to discover that first CO2 fixation product is 3-carbon organic compound (3-
phosphoglyceric acid) or PGA. Later on, a new compound was discovered which contain 4-
carbon called Oxaloacetic Acid (OAA). On the basis of number of carbon atoms in first stable
product they are named C3 and C4 pathway.

Calvin cycle can be described under three stages: carboxylation, reduction and regeneration.

1. Carboxylation is the fixation of CO2 into 3-phosphoglyceric acid (3-PGA).

Carboxylation of RuBP occurs in presence of enzyme RuBP carboxylase
(RuBisCO) which results in the formation of two molecules of 3-PGA.

2. Reduction is series of reaction that leads to formation of glucose. Two molecules of ATP
and two molecules of NADPH are required for reduction of one molecule of CO2. Six
turns of this cycle are required for removal of one molecule of Glucose molecules from
pathway.
3. Regeneration is the generation of RuBP molecules for the continuation of cycle. This
process requires one molecules of ATP.

Fig- Calvin Cycle/ C3 Cycle

 For every molecule of CO2 entering the Calvin Cycle, 3 molecules of ATP and 2
molecules of NADPH is required. To make one molecules of glucose 6 turns of cycle is
completed so total energy molecule required is

In Out
Six One glucose
18 ATP 18 ADP

12 NADPH 12 NADP
C4 pathway/Hatch Slack Pathway:

 This pathway was worked out by Hatch and Slack (1965, 1967), mainly operational in
plants growing in dry tropical region like Maize, Sugarcane, Sorghum etc.
 In this pathway first stable product is a 4-carbon compound Oxaloacetic acid (AAO) so
called as C4 pathway.
 C4 plants have Kranz Anatomy (vascular bundles are surrounded by bundle sheath cells
arranged in wreath like manner), characterized by large no of chloroplast, thick wall
impervious to gases and absence of intercellular spaces.
 The primary CO2 acceptor is a 3-carbon molecule Phosphoenol pyruvate present in
mesophyll cells and enzyme involved is PEP carboxylase.
 OAA formed in mesophyll cell forms 4-carbon compound like malic acid or aspartic acid
which is transported to bundle sheath cells.
 In bundle sheath cell, it is broken into CO2 and a 3-carbon molecule. The 3-carbon
molecule is returned back to mesophyll cells to form PEP.
 The CO2 molecules released in bundle sheath cells enters the Calvin cycle, where enzyme
RuBisCO is present that forms sugar.
Photorespiration:
 It is a light dependent process of oxygenation of RuBP and release of carbon dioxide by
photosynthetic organs of plants.
 Photorespiration decreases the rate of photosynthesis when oxygen concentration is increased
from 2-3% to 21%.
 Presence of light and higher concentration of Oxygen results in the binding of RuBisCO
enzyme with O2 to form.
RuBisCO + PGA + phosphoglycolate

This pathway involves Chloroplast, Peroxisome and Mitochondria. Photorespiration do not

occur in C4 plants.

C3 plants C4 plants
 The leaves do not have Kranz anatomy.  The leaves show Kranz anatomy in
 Photorespiration occurs. leaves.
 RuBisCO is the first acceptor of CO2.  Photorespiration does not occur.
  PGA is the first stable product.  PEP is the first acceptor of CO2.
 Plants are adapted to all climates.  OAA is the first stable product.
 Mesophyll cells perform complete  Plants are adapted to tropical climate.
photosynthesis.  Mesophyll cells perform only initial
fixation.

Factors affecting photosynthesis:

 The rate of Photosynthesis depends upon both internal as well as external factors.
 Internal factors like the number, size, age, Orientation of leaves, chloroplast etc., and Internal
like CO2 concentration, Temperature, water etc.
 When several factors affect any biochemical process, Blackman's (1905) Law of Limiting
Factors come into effect i.e. In case a chemical process is influenced by more than one factor
at that point its rate will be decided by the factor which is closest to its minimal value which
is called Limiting factor. It's the figure which specifically influences the method if the
amount is changed.

1. Light-
 Light is used as 10% of incident sunlight. At low intensity, it's directly proportional to
CO2 fixation. At higher intensity, other factors become limiting.
 At low light intensity, when there is no gaseous exchange in photosynthesis, it is called
light compensation point. As the light intensity increases, the rate of photosynthesis also
increases.
 The light intensity at which a plant can achieve maximum amount of photosynthesis is
called light saturation point.
 An increase in light beyond a limit causes the breakdown of chlorophyll and thus a
decrease in Photosynthesis.
2. Carbon dioxide concentration–
 It's the major limiting factor as carbon dioxide is very low in the atmosphere that is
0.03% to 0.04%, increase in concentration up to 0.05% can cause an increase in the rate
of photosynthesis.
 Increase in CO2 concentration increases rate of photosynthesis in most C3 plants.
 When CO2 concentration is reduced, there comes a point at which illuminated plant parts
stop absorbing CO2 from their environment. It is known as CO2 compensation point or
threshold value. At this value, CO2 fixed in photosynthesis is equal to CO2 evolved in
respiration and photorespiration.

3. Temperature-
 It does not influence the rate of photosynthesis directly but at higher temperature enzyme
activity is inhibited due to denaturation of enzymes which affect the dark reaction.
 The dark reactions being enzymatic are temperature controlled. C4 plants respond to
higher temperatures and they also show higher rates of photosynthesis whereas C3 plants
are found to show lower temperature optimum.
 When temperature is increased from minimum to optimum, the rate of photosynthesis
doubles for every 10˚C rise in temperature.
 Above the optimum temperature, the rate of photosynthesis shows an initial increase for
short duration but later declines.

4. Water–
 Water stress causes the stomata to close hence reducing the carbon dioxide availability
and it makes leaves wilt thereby reducing surface area.
 Due to increase in amount of water, rate of photosynthesis does not increase
proportionally as after saturation no more water is required during photosynthesis.
Blackman’s Law of Limiting Factors states:
If a chemical process is affected by more than one factor, then its rate will be determined by the
factor which is nearest to its minimal value: it is the factor which directly affects the process if
its quantity is changed.