Chapter 13 - PHOTOSYNTHESIS IN HIGHER PLANTS

Photosynthesis: Photosynthesis is an enzyme regulated anabolic process of manufacture of organic

compounds inside the chlorophyll containing cells from carbon dioxide and water with the help of sunlight as
a source of energy.

Historical Perspective:

 Joseph Priestley (1770): Showed that plants have the ability to take up CO2 from atmosphere and
release O2.

Priestly observed that a candle in a closed space – a bell jar, soon gets extinguished. Similarly, a
mouse would soon suffocate in a closed space. He concluded that a burning candle or an animal that
breath air, both somehow, damage the air. But when he placed the mint plant in the same bell jar, he
found that the mouse stayed alive and the candle continued to burn. Priestly hypothesized as
follows: plants restore to the air whatever breathing animals and burning candles remove.

 Jan Ingenhousz (1779): Release of O2 by plants was possible only in sunlight and only by the green
parts of plants.
 Ingenhousz in an elegant experiment with an aquatic plant showed that in bright sunlight, small
bubbles were formed around the green parts while in the dark they not. Later indentified these
bubbles to be oxygen. Hence he showed that it is only green part of the plants that could release
oxygen.
 Julius Von Sachs (1854): He showed that green substance in plants chlorophyll is located in special
bodies called chloroplast with in plant cells. He found that the green part in plants is where glucose is
made and that the glucose is usually stored as starch.
 Engelmann (1843 – 1909) – using a prism he split light into special components and then illuminated
a green alga, Cladophora, placed in suspension of aerobic bacteria. The bacteria were used to detect
the sites of oxygen evolution. He observed that the bacteria accumulated mainly in the region of blue
and red light of the split spectrum.The first action spectrum was described. It resembles roughly the
absorption of spectra of chlorophyll a and b

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By nineteenth century the key features of plant photosynthesis were known, namely, that plants could use
light energy to make carbohydrates from carbon dioxide and water. The equation was understood as

CO2 + H2O ------LIGHT---------- (CH2O) + O2

Later Cornelius van Neil (1897-1985) who based on his studies of purple and green bacteria demonstrated
that photosynthesis is essentially a light dependent reaction in which hydrogen from a suitable oxidizable
compound reduces carbon dioxide to carbohydrates. This can be expressed by

H2A + CO2 --------LIGHT-----------2A + CH2O + H2O

In green plants H2O is the hydrogen donor and is oxidized to O2, some organisms do not release O2 during
photosynthesis like in bacteria since it uses H2S, oxidation product is sulphur or sulphate depending on the
organism and not O2.

Hence he inferred that oxygen evolved by the green plant comes from H2O not from CO2. This was later
proved by using radio isotopic techniques.

Hence the correct equation of photosynthesis is therefore

CO2 + 12H20 ------------LIGHT------------ C6H12O6 + 6H2O + 6O2

Site for photosynthesis:

 Photosynthesis takes place only in green parts of the plant, mostly in leaves.
 Within a leaf, photosynthesis occurs in mesophyll cells which contain the chloroplasts.
 Chloroplasts are the actual sites for photosynthesis.
 The thylakoids in chloroplast contain most of pigments required for capturing solar energy to initiate
photosynthesis.
 The membrane system (grana) is responsible for trapping the light energy and for the synthesis of
ATP and NADPH. Hence it is called as light reaction, since it is directly dependent on light.
 Biosynthetic phase (dark reaction) is carried in stroma. It is not dependant on light; hence it is called
as dark reaction.

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(Study structure of chloroplast – fig 13.2 NCERT text book)

Pigments involved in photosynthesis:

Leaf pigments can be separated through paper chromatography. A chromatographic separation of leaf
pigments shows that the color we see in leaves is not due to a single pigment but due to four pigments as
follows

 Chlorophyll a: (Bright or blue green in chromatograph). Major pigment, act as reaction centre,
involved in trapping and converting light into chemical energy.
 Chlorophyll b : (Yellow green)
 Xanthophylls : (Yellow)
 Carotenoid : (Yellow to yellow-orange)

 Pigments are substances that have an ability to absorb light, at specific wavelengths.
 Fig 13.3a shows the ability of chlorophyll a pigment to absorb lights of different wavelength.
 Visible spectrum of light as VIBGYOR, with different wavelength.
 Fig 13.3b the graph shows wave length at which maximum absorption of chlorophyll a ie in
blue and red regions, also shows higher rate of photosynthesis.
 Fig 13.3c shows the overlapping between the absorption spectrum of chlorophyll a and action
spectrum of photosynthesis.
 These graph, together shows that most of the photosynthesis takes place in the blue and red
regions of the spectrum. Some photosynthesis does takes place at the other wavelengths of
the visible spectrum.
 Thus chlorophyll is the major pigment responsible for trapping light, other thylakoid pigments
like chlorophyll b, xanthophylls and carotenoids, which are called accessory pigments also
absorb light and transfer the energy to chlorophyll a, they not only enable a wider range of
wavelength of incoming light to be utilized for photosynthesis but also protect chlorophyll a
from photooxidation.

What is light reaction?

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 Light reactions or the ‘Photochemical ‘phase includes light absorption, splitting of water, evolution of
oxygen and formation of high energy compound like ATP and NADPH.
 Several protein complexes are involved in the process. The pigments are organized into two
photochemical Light Harvesting Complexes (LHC) within the photosystem I (PSI) and photosystem II
(PSII).
 The LHC are made up of hundreds of pigment molecules bound to proteins.
 Each photosystem has all the pigments except one molecule of chlorophyll ‘a’ forming a light
harvesting system (antennae). These pigments help to make photosynthesis more efficient by
absorbing different wavelengths of light.
 The single chlorophyll a molecule forms the reaction center. The reaction is different in both the
photosystems.
 In Photosystem I (PSI) the reaction center is Chlorophyll ‘a’ has an absorption peak at 700 nm,
hence referred as P700.
 While in Photosystem II (PSII) it has absorption peak at 680 nm, and is called P680.

Process of photosynthesis:

 It includes two phases - Photochemical phase and biosynthetic phase.

 Photochemical phase (Light reaction): This phase includes - light absorption, splitting of water,
oxygen release and formation of ATP and NADPH.
 Biosynthetic phase (Dark reaction): It is light independent phase, synthesis of food material
(sugars).

 Simultaneously electron released from photosystem-I is accepted by electron acceptor.

 Electron hole created in PS-I is filled up by the electron from PS-II.
 Electron from PS-I passed down hill and reduce NADP into NADPH+ + H+.

Photolysis of water:

 PS-II loose electrons continuously, filled up by electrons released due to photolysis of water.
 Water is split into H+, (O) and electrons in presence of light and Mn2+ and Cl-.
 This also creates O2 the bi-product of photosynthesis.

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 Photolysis takes place in the vicinity of the PS-II.
 2H2O → 4H+ + O2 + 4e-.

Photophosphorylation:

 The process of formation of high-energy chemicals (ATP and NADPH) in the presence of
sunlight.

Cyclic photophosphorylation: The electron transport:

 In photosystem centre chlorophyll a absorbs 680 nm wavelength of red light causing electrons to
become excited and release two electrons from the atomic nucleus.
 These electrons are accepted by primary electron acceptor i.e. ferredoxin.
 The electron from the feredoxin passed to electron transport system consisting cytochromes.
 The electron moved in downhill in terms of redox potential by oxidation-reduction reactions.
 Finally the electron reached photosystem-I.

 Only PS-I works, the electron circulates within the photosystem.

 It happens in the stroma lamellae (possible location) because in this region PS-II and NADP
reductase enzyme are absent.
 Hence only ATP molecules are synthesized.

Non Cyclic photophosphorylation:

 Two photo systems work in series – First PSII and then PSI.
 The electron released by PS I is accepted by electron acceptor and used to convert NADP TO NADPH.
 PS I get back the electron from PS II, during downhill reaction it produces one ATP.
 PSII gets back the electron from photolysis of water.
 These two photosystems are connected through an electron transport chain (Z. Scheme).
 PSI and PSII are found in lamellae of grana, hence this process is carried here.

Chemiosmotic Hypothesis :

 Chemiosmosis hypothesis explain the mechanism of ATP synthesis in chloroplast.

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 In photosynthesis, ATP synthesis is linked to development of a proton gradient across a membrane.

a) Splitting of water takes place in the inner side of the membrane. The protons or hydrogen ions that
are produced by the splitting of water accumulate within lumen of the thylakoids.
b) As the electron moves through the photosystem, protons are transported across the membrane.
This happens because the primary acceptor of electron which is located towards the outer side of
the membrane transfers its electron not to an electron carrier but to an H carrier. Hence the
molecule removes a proton from the stroma while transporting an electron. When this molecule
passes on it’s to the electron carrier on the inner side of the membrane, the proton is released into
the inner side or the lumen of the membrane.
c) NADP reductase enzyme is located on the stroma side of the membrane, along with electrons that
come from the acceptor of electrons of PS1, protons are necessary for the reduction of NADP to
NADPH + H+. these protons are also removed from the stroma.

 Hence with in the chloroplast, protons in stroma decrease in number, while in the lumen there is
accumulation of protons. This creates proton gradients across the thylakoid membrane as well as a
measurable decrease in pH in the lumen.
 This gradient is important because it is the breakdown of this gradient that leads to the synthesis of
ATP. The gradient is broken down due to the movement of protons across the transmembrane
channel of CF0 of the ATP synthase.
 The ATP synthase enzyme consists of two parts one called CF 0 is embedded in the thylakoid
membrane and forms a transmembrane channel that carries out the facilitated diffusion of protons
across the membrane. The other portion is called CF1 and protrudes outside on the outer surface of
the thylakoid membrane on the side that faces the stroma. The breakdown of the gradient provides
enough energy to cause conformational change in the CF1 particle of the ATP synthase, which makes
the enzyme synthesize several molecules of energy packed ATP.
 Chemiosmosis requires a membrane, a proton pump, a proton gradient and ATP synthase, energy is
used to pump protons across a membrane to create gradient or high concentration of protons within
the thylakoid lumen. ATP synthase has a channel that allows diffusion of protons back across the
membrane, this releases enough energy to activate ATP synthase enzyme that catalyses the
formation of ATP.

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 Along with the NADPH produced by the movement of electrons, the ATP will be used immediately in
the biosynthetic reaction taking place in the stoma, responsible for fixing CO2 , and synthesis of
sugars.

Biosynthetic phase in C3 plants:

 ATP and NADH, the products of light reaction are used in synthesis of food. The first
CO2 fixation product in C3 plant is 3-phosphoglyceric acid or PGA.
 In some other plants the first stable product is an organic acid called oxaloacetic acid a 4-C compound
hence is called C4 plants.

The Calvin cycle:

 The CO2 acceptor molecule is RuBP (Ribulose bis phosphate).

 The cyclic path of sugar formation is called Calvin cycle on the name of Melvin Calvin, the discoverer
of this pathway. Calvin cycle proceeds in three stages:
1. Carboxylation :
o Carboxylation is the fixation of CO2 into a stable organic intermediate.
o CO2 combines with Ribulose 1, 5 bisphosphate to form two molecules’ of 3 PGA in the presence
of RuBisCO enzyme ( Ribulose bis phosphate carboxylase oxygenase)

2. Reduction :

o These are a series of reactions that lead to the formation of glucose.

o 2 molecules of ATP for phosphorylation and two of NADPH for reduction per CO2 molecule fixed.
o The fixation of six molecules of CO2 and 6 turns of the cycle are required for the formation of one
molecule of glucose.

3. Regeneration :

o Regeneration of the CO2 acceptor molecule RuBP is crucial if the cycle is to continue
uninterrupted.
o The regeneration steps required one ATP for phosphorylation to form RuBP.

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  Hence for every CO2 molecule entering the Calvin cycle, 3 molecules of ATP and 2 molecules of
NADPH are required.

In Out
Six carbon dioxide One glucose
18 ATP 18 ADP
12 NADPH 12 NADP

The C4 pathway

 Plants that are adapted to dry tropical regions have C4 pathway.

 It is seen in monocot and some dicots.
 C4 Plants have special leaf anatomy, they can tolerate higher temperatures, they show a response to
high light intensities, and they lack a process called photorespiration and have greater productivity
of Biomass.
 In leaf anatomy of monocots particularly large cells are found around the vascular bundles of the C 4
plants called bundle sheath cells and the leaves which have such anatomy is called kranz anatomy.
 ‘Kranz’ means ‘wreath’ and is a reflection of the arrangement of cells.
 The bundle sheath cells may form several layers around the vascular bundles; they are characterized
by having large number of chloroplasts, thick walls impervious to gaseous exchange and no
intercellular space. Ex – Maize, Sorghum.

 The primary CO2 acceptor is a 3 carbon molecule phosphoenol pyruvate (PEP) and is present in
mesophyll cells. The enzyme responsible for this fixation is PEP carboxylase or PEPcase.
 The enzyme RuBisCO is absent in mesophyll cells.
 The C4 acid OAA is formed in the mesophyll cells.
 It then forms other 4 carbon compounds like malic acid or aspartic acid in the mesophyll cells itself,
which are transported to the bundle sheath cells.
 In the bundle sheath cells these C4 acids are broken down to release CO2 and a 3-carbon molecule
(phosphoenol pyruvate).
 The 3- carbon molecule is transported back into mesophyll where it is converted back into PEP again,
Thus completing the cycle.
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  The CO2 released in the bundle sheath cells enters the C3 or Calvin cycle, a pathway common to all the
plants.
 RuBisCO is present in bundle sheath cells but lack PEP case in bundle sheath cells.
 Thus, the basic pathway that results in the formation of sugars is the Calvin pathway. It is common
for both C3 and C4 plants.

Photorespiration:

 In the first step of Calvin cycle RUBP combines with CO2 to two molecule of PGA that is catalysed by
RuBisCo ( Ribulose bisphosphate carboxylase – oxygenase)
 RuBP + CO2 -------------- 2 X 3 PGA.
 RuBisCO has affinity for CO2 than O2
 In C3 plants some O2 binds with RuBisCo and hence CO2 fixation is decreased.
 In PHOTORESPIRATION - RuBP instead of being converted to 2 molecules of PGA, binds with
O2 to form one molecule of PGA and phosphoglycolate. This is called photorespiration.
 In the photo respiratory pathway there is neither synthesis of sugar, nor of ATP. Rather it results in
the release of CO2 with utilization of ATP.
 In the photo respiratory pathway there is no synthesis of ATP or NADPH.
 Therefore photorespiration is a wasteful process.
 In C4 plants photorespiration does not occur. This is because they have a mechanism that increases
the concentration of CO2 at the enzyme site. This takes place when C4 acid from mesophyll is
broken down in the bundle sheath cells to release CO2.
 This results in increasing the intercellular concentration of CO2. In turn this ensures that the RuBisCo
functions as a carboxylase minimizing the oxygenase activity.

Factors effecting photosynthesis

The rate of photosynthesis is very important in determining the yield of crop plants.

Photosynthesis is under influence of several factors, both internal (plant) and external.

The internal (plant factors) include the number, size, age and orientation of leaves, mesophyll cells and
chloroplasts, internal CO2 concentration and the amount of chlorophyll.

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The plant or internal factors are dependent on the genetic components and the growth of the plant.

The external factors would include the availability of sunlight, temperature, CO2 concentration and water.

As a plant photosynthesises all these factors will simultaneously affect photosynthesis or CO2 fixation.

Usually one factor is the major cause or is the one that limits the rate. Hence, at any point the rate will be
determined by the factor available at sub-optimal levels.

When several factors affect any (bio) chemical process, Blackman’s (1905) Law of limiting factors comes into
effect.

Law of Limiting Factors:

 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.

1. For example, despite the presence of a green leaf and optimal light and carbon dioxide conditions the
plant may not photosynthesize if temperature is low. This leaf if given optimal temperature will start
photosynthesizing.
2. For example in cloudy day the rate of photosynthesis will be controlled only by light.

External or environmental factors

1. Light:

• The influence of light depends on the quality, light intensity and the duration of exposure to light.
Maximum photosynthesis takes place in red and blue light.
• There is a linear relationship between incident light and carbon dioxide fixation rates at low light
intensities.
• At higher light intensities gradually the rate does not show further increase as other factors become
limiting.

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 • It is note that 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.
• Increase in light beyond a point causes the breakdown of chlorophyll and decrease in photosynthesis.
• Higher intensity of light results in bleaching or photoxidation of pigments which is called as
solarization.

2. Carbon dioxide concentration

 Carbon dioxide is the major limiting factor for photosynthesis.
 The concentration of CO2 is very low in the atmosphere (between 0.03 and 0.04 percent). Increase in
CO2 concentration upto 0.05 percent can cause increase in CO2 rates. Beyond this levels can become
damaging over longer periods.
 The C3 and C4 plants respond differently to CO2 concentrations.
 At low light conditions neither group will respond to high CO2 conditions.
 At high light intensities, both C3 and C4 show increase in the rates of photosynthesis.
 Important to note is C4 Plants show saturation at 360 microperlitre while C3 plants responds to
increase CO2 concentration and saturation is seen only beyond 450 microperlitre. Thus current
availability of carbon dioxide is limiting to the C3 plants.
 Since C3 plants respond to higher CO2 concentration by showing increased rates of photosynthesis
leading to higher productivity has been used in green house crops such as tomatoes and bell pepper.
Hence they are allowed to grow in CO2 enriched atmosphere that leads to higher yields.

3. Temperature:
 The rate increase with an increase in temperature up to about 400C. It is known as optimum
temperature. The rate of photosynthesis is maximum at an optimum temperature which may be
around 25-300C for mesophytes and 35-400 C for xerophytes. Beyond the optimum temperature,
the rate of photosynthesis decreases, the enzymes get denatured.
 The C4 plants respond to higher temperature and show higher rate of photosynthesis while C3
plants have a much lower temperature as optimum.
 The temperature optimum for photosynthesis of different plants also depends on habitat that they
are adapted to.
 Tropical plants have a higher temperature optimum than the plants adapted to temperate climates.

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 4. Water:
 Water is important for light reaction. The entire process of photosynthesis depends on the
availability of water; hence it is directly proportional to photosynthesis.
 Water stress causes the stomata to close hence reducing the CO2 availability.
 Besides water stress also makes leaves wilt, thus reducing the surface area of the leaves and their
metabolic activity also, hence reduces the yield.

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