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- Photosynthesis is a physico-chemical process by which - Ultimately, all living forms depend on sunlight for energy.
green plants use light energy (solar energy) to synthesise Importance of Photosynthesis
organic compounds. So they are autotrophs. • It is the primary source of all food on earth.
- It is the basis of life on earth. • It releases oxygen into the atmosphere.

EXPERIMENTS RELATED WITH PHOTOSYNTHESIS

1. Variegated leaf experiment Experiments by Julius von Sachs (1854)
- Take a variegated leaf (or leaf partially covered with black - He proved that
paper) that was exposed to light. o Glucose is produced when plants grow and it is usually
- Test the leaves for starch. It shows that photosynthesis stored as starch.
occurs only in green parts of the leaves in presence of light. o Chlorophyll is located in special bodies (chloroplasts).
2. Half-leaf experiment o Glucose is made in the green parts of plants.
- A part of a leaf is enclosed in a test tube containing KOH Experiments by T.W Engelmann (1843 – 1909)
soaked cotton (which absorbs CO2). - He split the light using a prism into its spectral components
- The other half of leaf is exposed to air. and illuminated a green alga (Cladophora) placed in a
- Place this setup in light for some time. suspension of aerobic bacteria.
- Test the leaf for presence of starch. Exposed part shows - The bacteria were used to detect the sites of O2 evolution.
positive for starch and portion in the tube shows negative. - He observed that the bacteria accumulated mainly in the
This proves that CO2 is required for photosynthesis. region of blue and red light of the split spectrum.
EARLY EXPERIMENTS - It was a first described action spectrum of photosynthesis.
It resembles the absorption spectra of chlorophyll a & b.
Experiments by Joseph Priestley (1770)
- By the middle of 19th century, it is discovered that plants
- Priestley performed experiments to prove the role of air in use light energy to make carbohydrates from CO2 & H2O.
the growth of green plants. - Empirical equation of the process of photosynthesis is
- He discovered oxygen in 1774.
- He observed that a candle burning in a closed bell jar gets
extinguished. Similarly, a mouse suffocated in closed jar. Where, [CH2O] represents a carbohydrate (e.g. glucose).
He concluded that a burning candle or a breathing animal Experiments by Cornelius van Niel (1897-1985)
damage the air. - Van Niel (microbiologist) conducted some studies in
- He placed a mint plant in the same bell jar. He found that purple and green bacteria.
the mouse stayed alive and the candle continued to burn. - He demonstrated that photosynthesis is a light-dependent
- He hypothesised that plants restore to the air whatever reaction in which hydrogen from an oxidisable compound
breathing animals and burning candles remove. reduces CO2 to carbohydrates.
Experiments by Jan Ingenhousz (1730-1799)
- He conducted the same experiment by placing in darkness
- In plants, H2O is the hydrogen donor and is oxidised to O2.
and sunlight.
- Purple & green sulphur bacteria use H2S as H-donor. So
- He showed that sunlight is essential to the plant for
the ‘oxidation’ product is sulphur or sulphate and no O2 is
purifying the air fouled by burning candles or animals.
produced.
- He repeated this experiment with an aquatic plant. It
- Thus, he inferred that the O2 evolved by the green plant
showed that in bright sunlight, small bubbles were formed
comes from H2O, not from CO2. This was later proved by
around green parts while in the dark they did not.
using radio isotopic techniques.
- Later he identified these bubbles to be of oxygen. Thus he
- Therefore overall correct equation for photosynthesis is:
showed that only the green part of plants release O2.

PHOTOSYNTHESIS: SITE AND PIGMENTS

- Photosynthesis occurs in green leaves & other green parts. - The membrane system traps light energy and synthesise
- Chloroplasts present in the walls of mesophyll cells of ATP and NADPH. It is called light reactions.
leaves. It helps to get optimum quantity of incident light. - In stroma, enzymatic reactions synthesize sugar, which in
- Chloroplast contains a membranous system. It consists of turn forms starch. It is called dark reactions (carbon
grana, stroma lamellae and matrix stroma. reactions). It does not mean that they occur in darkness or
- Each granum is a group of membrane-bound sacs called that they are not light dependent.
thylakoids (lamellae). They contain leaf pigments.

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Graph showing
action
spectrum of
photosynthesis

PIGMENTS INVOLVED IN PHOTOSYNTHESIS

Graph showing
- Pigments are substances that have ability to absorb light at
action spectrum of
specific wavelengths. photosynthesis
- Chromatography shows the following leaf pigments: superimposed on
absorption
o Chlorophyll a (bright or blue green in chromatogram) spectrum of
o Chlorophyll b (yellow green) chlorophyll a
Accessory
o Xanthophylls (yellow)
pigments
o Carotenoids (yellow to yellow-orange)
- Functions of accessory pigments: Photosystems
o They absorb light at different wavelength and transfer - Pigments are organised into two Photosystems called
the energy to chlorophyll a. Photosystem I (PSI) & Photosystem II (PSII). These are
o They protect chlorophyll a from photo-oxidation. named in the sequence of their discovery.
- The absorption spectrum & action spectrum coincide - Each photosystem has a chlorophyll a and accessory
closely showing that photosynthesis is maximum at the pigments bound by proteins.
blue & red regions of the spectrum. - All pigments (except one molecule of chlorophyll a) form
- The graphs also show that chlorophyll a is the chief a light harvesting
pigment associated with photosynthesis. complex (LHC or
antennae).
- Single chlorophyll a acts
Graph showing as reaction centre.
absorption
spectrum of
- In PS I, the reaction
chlorophyll a, b centre absorbs light at
& carotenoids 700 nm, and so called
P700.
- In PS II, the reaction
centre absorbs light at 680 nm, and so called P680.

LIGHT REACTION (PHOTOCHEMICAL PHASE)

- Light reactions include light absorption, water splitting,
oxygen release and formation of ATP & NADPH (high-
energy chemical intermediates).
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 to a chain of electrons
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.
- Simultaneously, electrons in PS I are also excited when
they receive red light of 700 nm and are transferred to Z scheme of light reaction
another accepter molecule having a greater redox potential. Splitting of Water (Photolysis)
- These electrons are moved downhill to a molecule of
- The water splitting complex in PS II is located on the inner
NADP+. As a result, NADP+ is reduced to NADPH + H+.
side of the thylakoid membrane.
- Transfer of electrons from PS II to PS I and finally downhill - Water is split into H+, [O] and electrons.
to NADP+ is called the Z scheme, due to its zigzag shape.
2H2O → 4H+ + O2 + 4e−
This shape is formed when all the carriers are placed in a
- So PS II can supply electrons continuously by replacing
sequence on a redox potential scale.
electrons from water splitting.

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- Thus PS II provides electrons needed to replace those of protons from the stroma for the following reasons:
removed from PS I. o Primary electron accepter is located towards the outer
- The protons (H+) are used to reduce NADP to NADPH. side of the membrane. It transfers its electron to an H
- Oxygen is liberated as a by-product of photosynthesis. carrier. So this molecule removes a proton from the
stroma while transporting an electron. When this
Photo-phosphorylation
molecule passes on its electron to the electron carrier
- The synthesis of ATP by cells (in mitochondria &
on the inner side of the membrane, proton is released
chloroplasts) is called phosphorylation.
into the lumen of the membrane.
- Photo-phosphorylation is the synthesis of ATP from ADP
o The NADP reductase enzyme is located on the stroma
in chloroplasts in presence of light.
side of the membrane. Along with electrons coming
- It occurs in 2 ways: Non- cyclic and Cyclic.
from PS I, protons are necessary to reduce NADP+.
a) Non-cyclic photo-phosphorylation These protons are also removed from the stroma.
- It occurs when the two photosystems work in a series, (first - Hence, protons in stroma are decreased but in lumen,
PS II and then PS I) through an electron transport chain as protons are accumulated. It creates a proton gradient across
seen in the Z scheme. the thylakoid membrane and decrease in pH in the lumen.
- Here, ATP & NADPH + H+ are synthesised.
- It is a non-cyclic process because the electrons lost by PS
II do not come back to it but pass 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
- Breakdown of proton gradient leads to synthesis of ATP by
stroma lamellae
membranes lack PS II and NADP reductase. ATP synthase enzyme.
- The ATP synthase consists of two parts:
- The electron does not pass on to NADP+ but is cycled back
to PS I complex through electron transport chain. o CF0: It is embedded in the membrane and forms a trans-
membrane channel. It carries out facilitated diffusion of
- Here, only ATP is synthesised (no NADPH + H+).
protons across the membrane to the stroma. It results in
- Cyclic photophosphorylation also occurs when only light
breakdown of proton gradient.
of wavelengths beyond 680 nm are available.
o CF1: It protrudes on the outer surface of the thylakoid
Chemiosmotic Hypothesis membrane. The energy due to breakdown of gradient
- It explains mechanism of ATP synthesis in chloroplast. causes a conformational change in the CF1 particle. It
- Chemiosmosis: Movement of ions across a semipermeable makes the enzyme to synthesise ATP molecules.
membrane. It occurs in chloroplast and mitochondria. - Energy is used to pump protons across a membrane, to
- Chemiosmosis needs a membrane, a proton pump, a proton create a gradient or a high concentration of protons within
gradient (across thylakoid membranes) and ATP synthase. the thylakoid lumen.
- Splitting of water occurs on the inner side of the membrane. - ATP synthase has a channel for the diffusion of protons
So the protons accumulate in the lumen of thylakoids. back across the membrane. This releases energy to activate
- As electrons move through the photosystems, protons are ATP synthase that catalyses formation of ATP.
transported across the membrane. It is due to the removal

DARK REACTION (BIOSYNTHETIC PHASE) - USE OF ATP & NADPH

- Products of light reaction are ATP, NADPH and O2. for some time, and then stops. If light is available, the
- Dark reaction is the use of ATP and NADPH to drive the synthesis starts again.
processes for the synthesis of food (sugars). - CO2 combines with H2O to form (CH2O)n or sugars.
- This phase does not directly depend on the light but is - CO2 assimilation during photosynthesis is 2 types:
dependent on the products of the light reaction. o C3 pathway: In this, first stable product of CO2 fixation
- It can be verified as follows: Immediately after light is a C3 acid (3-phosphoglyceric acid - PGA). Melvin
becomes unavailable, the biosynthetic process continues Calvin discovered this using 14C in algal photosynthesis.

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o C4 pathway: In this, first stable product is oxaloacetic - The bundle sheath cells may form several layers around
acid (OAA), a 4-carbon (C4) organic acid. the vascular bundles.
C3 PATHWAY (CALVIN CYCLE) - They have large number of chloroplasts, thick walls
impervious to gas exchange and no intercellular spaces.
- It occurs in all photosynthetic plants (C3 or C4 pathways).
- It has 3 stages: carboxylation, reduction and regeneration. Steps of Hatch and Slack Pathway
- Primary CO2 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 C4 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.

1. Carboxylation of RuBP
- RuBP (ribulose bisphosphate - a 5-carbon ketose sugar)
is the primary CO2 acceptor.
- It is the most crucial step. CO2 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).
- RuBisCO is the most abundant enzyme in the world.
2. Reduction
- It is a series of reactions leading to the glucose formation.
- Here, 2 ATP molecules for phosphorylation and two of
- In the bundle sheath cells, C4 acids are broken down to
NADPH for reduction per CO2 molecule are used.
release CO2 and a C3 molecule.
- Fixation of 6 CO2 molecules and 6 turns of the cycle are
- The C3 molecule is transported back to mesophyll where it
needed to remove one glucose molecule from the pathway.
is converted to PEP again.
3. Regeneration of RuBP - The released CO2 enters the C3 pathway.
- It is crucial for continuation of the cycle. - Bundle sheath cells are rich in RuBisCO, but lack
- It requires one ATP for phosphorylation to form RuBP. PEPcase. Thus C3 pathway is common to C3 & C4 plants.
- Hence for every CO2 molecule, 3 ATP molecules and 2 C4 plants are special because:
NADPH are required. o They have a special type of leaf anatomy (Kranz).
- It is probably to meet this difference in number of ATP and o They tolerate higher temperatures.
NADPH used in the dark reaction that the cyclic o They show a response to highlight intensities.
phosphorylation takes place. o They lack photorespiration.
- To make 1 glucose molecule, 6 turns of the cycle are needed. o They have greater productivity of biomass.
In Out
What does go in and PHOTORESPIRATION
6 CO2 1 glucose
come out of the Calvin - In Calvin pathway, RuBP combines with CO2.
18 ATP 18 ADP
cycle?
12 NADPH 12 NADP
- Active site of RuBisCO can bind to CO2 & O2 – so the name.
C4 PATHWAY (HATCH & SLACK PATHWAY)
- RuBisCO has a greater affinity for CO2 than for O2. This
- It is present in plants adapted to dry tropical regions. binding is competitive. Relative concentration of O2 and
- They also use C3 pathway as main biosynthetic pathway. CO2 determines which one will bind to the enzyme.
- The large cells around the vascular bundles of the C4 plants - In C3 plants, some O2 bind to RuBisCO. Hence CO2 fixation
are called bundle sheath cells. Such anatomy is called is decreased. Here RuBP binds with O2 to form one
‘Kranz’ anatomy (‘Kranz’ = ‘wreath’).
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molecule of phosphoglycerate and phosphoglycolate. This This takes place when C4 acid from the mesophyll is broken
pathway is called photorespiration. down in the bundle cells to release CO2. This minimises the
- In this, there is no synthesis of sugars, ATP and NADPH. oxygenase activity of RuBisCO.
Hence photorespiration is a wasteful process. Rather it - Due to the lack of photorespiration, productivity and yields
causes the release of CO2 by using ATP. are better in C4 plants. Also, these plants show tolerance to
- In C4 plants, photorespiration does not occur because higher temperatures.
they can increase CO2 concentration at the enzyme site.
Differences between C3 and C4 plants
C3 plants C4 plants
1. Photosynthesis occurs in mesophyll cells. In mesophyll and bundle sheath cells.
2. Kranz anatomy is absent. Present.
3. RuBP is the primary CO2 acceptor. PEP is the primary CO2 acceptor.
4. 3-PGA, a 3-C compound is the first stable product. OAA, a 4-C compound is the first stable product.
5. Chloroplasts are of only one type (granal). Dimorphic (granal in mesophyll and agranal in bundle sheath).
6. Photorespiratory loss is high. Photorespiration is absent or negligible.
7. High CO2 compensation point (25-100 µl. CO2 l-1). Low CO2 compensation point (0-10 µl. CO2 l-1).
8. Optimum temperature for photosynthesis is about 25oC. About 35oC - 45oC.
9. Photosynthetically less efficient and productivity low. Photosynthetically more efficient and productivity high.
10. E.g. Rice, wheat, bean, potato. E.g. Maize, sugarcane, amaranth, sorghum.

FACTORS AFFECTING PHOTOSYNTHESIS

- Internal (plant) factors: The number, size, age and Carbon dioxide Concentration
orientation of leaves, mesophyll cells and chloroplasts, - CO2 is the major limiting factor for photosynthesis.
internal CO2 concentration and amount of chlorophyll. - CO2 concentration is very low in the atmosphere (0.03-
Plant factors depend on the genes and growth of the plant. 0.04%). Increase up to 0.05% cause increase in CO2
- External factors: Sunlight, temperature, CO2 fixation rates. Beyond this level can become damaging
concentration and water. over longer periods.
- Blackman’s Law of Limiting Factors (1905): “If a - At low light, C3 and C4 plants do not respond to high CO2.
biochemical process is affected by more than one factor, At high light, they show increased rate of photosynthesis.
its rate is determined by the factor nearest to its minimal - C4 plants show saturation at about 360 µlL-1.
value: it is the factor which directly affects the process if - C3 plants respond to increased CO2 concentration and
its quantity is changed.” saturation is seen only beyond 450 µlL-1. Thus, current
- E.g. a plant with green leaf, optimal light & CO2 conditions availability of CO2 levels is limiting to the C3 plants.
may not photosynthesize if the temperature is very low. If - Due to response to higher CO2 concentration, C3 plants
optimal temperature is given, it will start photosynthesis. show increased photosynthesis and higher productivity.
Light This fact is used for some greenhouse crops (tomatoes, bell
- Light quality, light intensity and duration of exposure pepper etc). They are grown in CO2 enriched atmosphere.
to light influence photosynthesis. Temperature
- There is a linear relationship between incident light and - Dark reactions, being enzymatic, are temperature controlled.
CO2 fixation rates at Influence of temperature on Light reactions is very less.
low light intensities. - The C4 plants respond to higher temperatures and show
- At higher light higher rate of photosynthesis.
intensities, the rate - C3 plants have a much lower temperature optimum.
does not show - The temperature optimum of plants also depends on their
further increase habitat. Tropical plants have a higher temperature optimum
because other factors than the plants adapted to temperate climates.
become limiting.
Water
- Light saturation
occurs at 10% of the full sunlight. Hence, except for plants - Water stress closes the stomata hence reduce the CO2
availability.
in shade or in dense forests, light is rarely a limiting factor
- Water stress also wilts leaves, thus reduce the surface area
in nature.
of the leaves and their metabolic activity.
- High increase in incident light breaks down chlorophyll. It
decreases photosynthesis.