13

PHOTOSYNTHESIS
KEYNOTES 2.0
By: Dr. Anand Mani

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 photosynthesis
Photosynthesis is a physico- Photosynthesis is the basis of
chemical process by which green life on earth.
plants use light energy (solar
energy) to synthesise organic Ultimately, all living forms
compounds. depend on sunlight for energy.

Importance of Photosynthesis

It is the primary source of all food on earth.

It releases oxygen into the atmosphere

EXPERIMENTS RELATED WITH PHOTOSYNTHESIS

1. Variegated leaf experiment

Take a variegated leaf (or leaf partially covered with black paper) that was exposed to
light.

Test the leaves for starch. It shows that photosynthesis occurs only in green parts of the
leaves in presence of light.

2. Half-leaf experiment
VON MOHL
A part of a leaf is enclosed in a test tube containing KOH soaked cotton (which absorbs
CO2 ).

The other half of the leaf is exposed to air.

Place this setup in light for some time.

Test the leaf for presence of starch. Exposed part tests positive for starch and portion
in the tube tests negative. This proves that CO 2is required for photosynthesis.

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 early experiments

1. Experiments by Joseph Priestley (1770)

Priestley performed experiments to prove the role

of air in the growth of green plants.

He discovered oxygen in1774.

He observed that a candle burning in a closed bell

jar gets extinguished. Similarly, a mouse soon
suffocated in a closed space. He concluded that a
burning candle or a breathing animal damage the air.

He placed a mint plant in the same bell jar. He found

that the mouse stayed alive and the
candle continued to burn.

He hypothesised that plants restore to the air

whatever breathing animals and burning
candles remove.

2. Experiments by Jan Ingenhousz (1730 - 1799)

• He conducted the same • He repeated this experiment

experiment by placing it once with an aquatic plant. It
showed that in bright sunlight,
in the dark and once in the
small bubbles were formed
sunlight.
around green parts while in the
dark they did not.
• He showed that sunlight is
essential to the plant for
• Later he identified these
purifying the air fouled by bubbles to be of oxygen. Thus
burning candles or animals. he showed that only the green
parts of plants release O2 .

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3. Experiments by Julius von Sachs (1854)

He proved that-
• Glucose is produced when plants grow and it is usually stored as starch.

• Chlorophyll is located in special bodies (chloroplasts).

• Glucose is made in the green parts of plants.

4. Experiments by T.W Engelmann (1843 – 1909)

• He split the light using a prism into its spectral components and illuminated a
green alga (Cladophora) placed in a suspension of aerobic bacteria.

• The bacteria were used to detect the sites of O 2 evolution.

• He observed that the bacteria accumulated mainly in the region of blue and red
light of the split spectrum.

• It was the first described action spectrum of photosynthesis. It resembles the

absorption spectra of chlorophyll a & b.

• By the middle of 19th century, it was discovered that plants use light energy to
make carbohydrates from C 2O & H O.
2
2

• Empirical equation of the process of photosynthesisis

• Where, [CH2 O] represents a carbohydrate (e.g., glucose).

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4. Experiments by Cornelius van Niel (1897 1985)

• Van Niel (microbiologist) conducted some studies in purple and green bacteria.

• He demonstrated that photosynthesis is a light-dependent reaction in which

hydrogen from a suitable oxidisable compound reduces CO 2 to carbohydrates.

• This can be expressed by:

• In green plants, H 2O is the hydrogen donor and is oxidised to O2 .

• Purple and green sulphur bacteria use H 2S as H-donor. So the ‘oxidation’ product
is sulphur or sulphate and no O 2 is produced.

• Thus, he inferred that the O2 evolved by the green plant comes from H 2O, not
from CO2 . This was later proved by using radioisotopic techniques.

• Therefore overall correct equation for photosynthesis is:

PHOTOSYNTHESIS: SITE AND PIGMENTS

• Photosynthesis occurs in green leaves & other

green parts.

• Chloroplasts are present in the of mesophyll cells

of leaves. It helps to get optimum
quantity of incident light.

• Chloroplast contains a membranous system. It

consists of grana, stroma lamellae and
fluid stroma.

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 • Each granum is a group of membrane-bound
sacs called thylakoids (lamellae). They contain
leaf pigments.

• The membrane system traps light energy

and synthesis ATP and NADPH. It is called
light reaction.

• In stroma, enzymatic reactions incorporate

CO2 into the plant for synthesizing sugar,
which in turn forms starch. It is called dark
reaction. It does not mean that they occur in
darkness or that they are not light dependent.

1. PIGMENTS INVOLVEDIN PHOTOSYNTHESIS

• Pigments are substances that have the ability to absorb

light, at specific wavelengths.
• Chromatography shows the following leaf pigments:

Chlorophyll a (bright or blue green in

chromatogram)

Chlorophyll b (yellow green)

Xanthophylls (yellow)

Carotenoids (yellow to yellow-orange)

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Functions of accessory pigments:
1.They absorb light at different wavelength and
transfer the energy to chlorophyll .

2. They protect chlorophyll a from photo-

oxidation.

3. The absorption spectrum & action spectrum

coincide closely showing that
photosynthesis is maximum in the blue & red
regions of the spectrum.

4. The graphs also show that chlorophyll a is the

chief pigment associated with photosynthesis

Photosystems

Pigments are organised into two Photosystems called Photosystem I

(PSI) & Photosystem II (PSII). These are named in the sequence of their
discovery.

Each photosystem has a chlorophyll a molecule and accessory

pigments bound to proteins.

All pigments (except

one molecule of
chlorophyll a) form a
light harvesting
complex (LHC or
antennae).

• Single chlorophyll a acts as reaction centre.

• In PS I, the reaction centre absorbs light upto 700 nm, and so called P700.

• In PS II, the reaction centre absorbs light upto 680 nm, and so called P680.

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 LIGHT REACTION (PHOTOCHEMICAL PHASE)

+
2e - + 2H + [0]

Light reactions include light absorption, water splitting, oxygen release

and formation of ATP & NADPH (high-energy chemical intermediates).

1. The Electron Transport

When PS II absorbs red light of 680 nm wavelength, electrons are excited

and transferred to an electron acceptor.

The electron acceptor passes them through a chain of electron transport

system consisting of cytochromes.

This movement of electrons is downhill, in terms of redox potential scale.

The electrons are transferred to the pigments of PS I.

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 Simultaneously, electrons in
PS I are also excited when
they receive red light of 700
nm and are transferred to
another accepter molecule
having a greater redox
potential.

These electrons are moved downhill to a molecule of NADP+. As a result, NADP

+ is reduced to NADPH +H+.

Transfer of electrons from PS II to PS I and finally downhill to NADP+ is called

the Z scheme, due to its zigzag shape. This shape is formed when all the carriers
are placed in a sequence on a redox potential scale.

2. Splitting of Water (Photolysis)

The water splitting complex in PS II is located on the inner side of the thy lakoid
membrane.

Water is split into H+ , [O] and electrons.

2H2O – 4H+ + O 2 + 4e−

These electrons are needed to replace the electrons that are moved from PS II

The protons (H +) are used to reduce NADP to NADPH.

Oxygen is liberated as a by-product of photosynthesis.

PS II provides electrons needed to replace those removed from PS I.

3. Photo-phosphorylation
• The synthesis of ATP by cells (in mitochondria &
chloroplasts) is called phosphorylation.
• Photo-phosphorylation is the synthesis of ATP
from ADP in chloroplasts in presence of light.
• It occurs in 2 ways: Non- cyclic and Cyclic.

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 a) Non-cyclic photo-phosphorylation

It occurs when the two photosystems work in a series,

(first PS II and then PS I) through an electron
transport chain as seen in the Z scheme.

Here, ATP & NADPH + H+ are synthesised.

It is a non-cyclic process because gets the electrons

lost by PS II do not come back to it but passed on to
NADP+
b) Cyclic photo - phosphorylation

It occurs in stroma lamellae when only PS I is

functional.

The electron is circulated within the photosystem

and the ATP synthesis occurs due to cyclic flow of
electrons.

The lamellae of grana have PS I & PS II. The stromal

lamellae membranes lack PS II and NADP reductase.

The electron does not pass on Here, only ATP is Cyclic photophosphorylation
to NADP+ but is cycled back synthesised (no also occurs when only light
to PS I complex through NADPH H+). of wavelengths beyond 680
electron transport chain. +
nm are available.

4. Chemiosmotic Hypothesis

It explains mechanism of ATP synthesis in chloroplast.

ATP synthesis is linked to development of a proton gradient across thylakoid

membranes.

Splitting of water occurs on the inner side of the membrane. So the protons
accumulate in the lumen of thylakoids.

As electrons move through the photosystems, protons are transported across

the membrane. It is due to the removal of protons from the stroma for the
following reasons:

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The primary electron accepter The NADP reductase enzyme is located on the
is located towards the outer stroma side of the membrane. Along with
side of the membrane and electrons coming from PS I, protons are
transfers its electron to an H necessary to reduce NADP+. These protons
carrier. So this molecule are also removed from the stroma.
removes a proton from the
stroma while transporting an
electron. When this molecule
passes on its electron to the Hence, protons in the stroma decrease in
electron carrier on the inner number, while in the lumen, protons are
side of the membrane, proton accumulated. This creates a proton gradient
is released into the lumen of across the thylakoid membrane and decrease
the membrane. in pH in the lumen.

Breakdown of proton gradient leads to

release of energy.

The gradient is broken down due to the

movement of protons across the
membrane to the stroma through the
trans-membrane channel of the F0 of the
enzyme ATPase.

The ATPase (ATP synthase) consists of two parts:

• F0 : It is embedded in the membrane and forms a trans - membrane channel that \
carries out facilitated diffusion of protons across the membrane.

• F1 : It protrudes on the outer surface of the thylakoid membrane. The energy due to
breakdown of gradient causes a conformational change in the F1 particle. It
makes the enzyme to synthesise ATP molecules.

Energy is used to pump ATPase has a channel that

Thus, chemiosmosis
protons across a membrane, allows diffusion of protons
requires a membrane,
to create a gradient or a back across the membrane.
a proton pump, a
high concentration of This releases energy to
proton gradient and
protons within the thylakoid activate ATPase enzyme that
ATPase.
lumen. catalyses formation of ATP.

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 DARK REACTION (BIOSYNTHETIC PHASE) USE OF ATP & NADPH

• This phase does not directly depend

• Products of light reaction are on light but is dependent on the
ATP, NADPH and O2. products of light reaction.

• Dark reaction is the use of ATP • It can be verified as follows:

and NADPH to drive the processes Immediately after light becomes
for the synthesis of food (sugars). unavailable, the biosynthetic process
continues for some time, and then
stops. If light is available, the
synthesis starts again.

• C 3 pathway: In this, first stable product

• CO 2 combines with H2O to form of CO 2 fixation is a C 3 acid (PGA). Melvin
(CH 2O)n or sugars. Calvin studied algal photosynthesis using
14c . He discovered that the first CO 2
• CO 2 assimilation during fixation product was 3-phosphoglyceric
photosynthesis is of 2 types : acid (PGA), a 3-carbon organic acid.

C 4 pathway: In this, first stable product is oxaloacetic acid (OAA), a 4-carbon

(C4 ) organic acid.

1. C3 PATHWAY (CALVIN CYCLE)

• It occurs in all photosynthetic plants (C3 or C4 pathways).
• It has 3 stages: carboxylation, reduction and regeneration.

1 Carboxylation of RuBP

RuBP (ribulose bisphosphate - a 5-carbon ketose sugar) is

the primary CO 2 acceptor.

It is the most crucial step. CO 2 is fixed by RuBP to two 3-

PGA in presence of the enzyme RuBP carboxylase.

Since this enzyme also has an oxygenation activity it is

called RuBP carboxylase-oxygenase (RuBisCO).

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 2

3ATPt2NADP
2 Reduction
• It is a series of reactions leading to the
glucose formation.

• Here, 2 ATP molecules for

phosphorylation and two of NADPH for
reduction per CO 2 molecule are used.

• Fixation of 6 CO 2 molecules and 6 turns

of the cycle are needed to remove one
glucose molecule from the pathway.

18ATP t I 2 NADPH

3 Regeneration of RuBP

• It is crucial for continuation of the cycle.

RUBISCO

• It requires one ATP for phosphorylation to form RuBP.

• Hence for every CO 2 molecule, 3 ATP molecules and 2 NADPH

molecule are required.

• It is probably to meet this difference in number of ATP and NADPH

used in the dark reaction that cyclic phosphorylation takes place.

• To make 1 glucose molecule, 6 turns of the cycle are needed

What goes in and comes out of the Calvin cycle?

In out
6 CO 2 1 glucose

18 ATP 18 ADP

12 NADPH 12 NADP

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2. C4 PATHWAY (HATCH & SLACK PATHWAY) 30ATP ILNADPH
• It is present in plants adapted to dry tropical
regions.

• They also use C pathway as main biosynthetic

pathway. The large cells around the vascular
bundles of the C4 plants are called bundle
sheath cells. Such anatomy is called ‘Kranz’
anatomy (‘Kranz’ = ‘wreath’).

• The bundle sheath cells may form several

layers around the vascular bundles.

• They have large number of chloroplasts, thick

walls impervious to gas exchange and no
intercellular spaces.

Steps of Hatch and Slack Pathway

Primary CO 2 acceptor is phosphoenol pyruvate (PEP) - a 3-carbon

molecule seen in mesophyll cells. The enzyme for this fixation is
PEP carboxylase (PEPcase).

• The mesophyll cells lack RuBisCO enzyme.

• The C 4 acid OAA is formed in the mesophyll cells.

• It then forms other 4-carbon acids like malic acid or aspartic acid. They are
transported to bundle sheath cells.

• In the bundle sheath cells, C4 acids are broken down to release CO2 and a C 3
molecule.

• The C 3 molecule is transported back to mesophyll where it is converted to

PEP again.

• The released CO 2 enters the C3 pathway.

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 Bundle sheath cells are rich in RuBisCO, but lack PEP case. Thus C 3 pathway is
common to C 3 & C4 plants. C 4 plants are special because:

• They have a special type of leaf anatomy (Kranz).

• They tolerate higher temperatures.
• They show a response to high light intensities.
• They lack photorespiration.
• They have greater productivity of biomass.

RuBisco PHOSPHO9LUCERATE3
3. PHOTORESPIRATION Oz Phosphoglucolate2

In Calvin pathway, RuBP combines with CO2 . RuBisCO has a greater

affinity for CO 2 than for O2
. This binding is
competitive. Relative
RuBisCO is the most abundant enzyme in the concentration of O 2 and
world. Its active site can bind to both CO 2 and CO2 determines which one
O 2 – hence the name. will bind to the enzyme.

P
if In C 3 plants, some O 2 binds
to RuBisCO. Hence CO 2
fixation is decreased.
Here RuBP binds with2 O
to form one molecule of
phosphoglycerate and
phosphoglycolate. This
pathway is called
photorespiration.

In this, there is no synthesis In C4 plants, photorespiration does not

of sugars, ATP and NADPH. occur because they can increase CO2
Hence photorespiration is a concentration at the enzyme site. This
wasteful process. Rather it takes place when C 4 acid from the
causes the release of CO2 mesophyll is broken down in the bundle
by using ATP. cells to release CO2 . This minimises the
oxygenase activity of RuBisCO.

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 Due to the lack of photorespiration, productivity and yields are better in4 C
plants. In addition, these plants show tolerance to higher temperatures
4

4. Differences between C3 and C4 plants

C 3 Plants C4 Plants
Photosynthesis occurs in mesophyll cells In mesophyll and bundle sheath cells

Kranz anatomy absent present

RuBP is the primary co2 acceptor PEP is the primary co 2 acceptor

3-PGA, a 3-c compound in the

first stable product
OOA, a 4-C compound is the first stable product

Chloroplast are of only one Dimorphic (granal in mesophyll and agranal in bundle
type (granal) sheath)

photo respiratory loss is high Photo respiration is absent or negligible

Optimum temprature for About 35O c-45 c

O

photosynthesis is about 25 Oc

Photosyntheticall less efficent and productivity Photosynthetically more efficent and

low productivity high

Eg. Rice, wheat, bean, potato Eg. Maize, sugarcane, amaranth, sorghum

tomato

FACTORS AFFECTING PHOTOSYNTHESIS

Internal (plant) factors: The number,

size, age and orientation of leaves,
External factors: Sunlight,
mesophyll cells and chloroplasts,
temperature, CO 2
internal CO2 concentration and
concentration and water.
amount of chlorophyll.

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 Blackman’s Law of Limiting Factors (1905): “If a biochemical process is affected
by more than one factor, its rate is determined by the factor nearest to its
minimal value: it is the factor which directly affects the process if
its quantity is changed.”
• E.g. a plant with green leaf, optimal light & CO2 conditions may not
photosynthesize if the temperature is very low. If optimal temperature is
given, it will start photosynthesis.

1. Light

Light quality, light intensity and duration of

exposure to light influences photosynthesis.

There is a linear relationship between

incident light andCO2 fixation rates at low
light intensities.

At higher light intensities, the rate does not

further show increase because other factors
become limiting.

Light saturation occurs at 10% of the full

sunlight. Hence, except for plants in shade or
in dense forests, light is rarely a limiting
factor in nature.

High increase in incident light breaks down

chlorophyll. It decreases photosynthesis.

2. Carbondioxide Concentration
• CO2 is the major limiting factor for
photosynthesis.

• CO concentration is very low in

2
the atmosphere (0.03 - 0.04%).
Increase in concentration up to
0.05% cause increase in CO2 fixation
rates. Beyond this the levels can
become damaging over longer
periods.

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 At low light, C3 and C4 plants do not
respond to high CO2 . At high light, they
show increased rate of photosynthesis.

• C4 plants show saturation at about 360

μlL-1

• C3 plants respond to increased CO 2

concentration and saturation is seen only
beyond 450 μlL-1. Thus, current availability
of CO 2 levels is limiting to the C3 plants.

Due to response to higher CO 2

concentration, C3 plants show
increased photosynthesis and
higher productivity. This fact is
used for some green house
crops (tomatoes, bell pepper
etc). They are grown in CO2
enriched atmosphere.

3. Temperature

Dark reactions, being enzymatic, are temperature controlled. Influence of temperature on

Light reactions is very less.

The C4 plants respond to higher temperatures and show higher rate of photosynthesis.

C3 plants have a much lower temperature optimum.

The temperature optimum of plants also depends on their habitat. Tropical plants have a
higher temperature optimum than the plants adapted to temperate climates.

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4. Water

• Water stress closes the stomata

hence reduce the CO 3 availability.

• Water stress also wilts leaves,

thus reduce the surface area of
the leaves and their metabolic
activity.

Additional Points

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