Course outline

I- Carbon nutrition (photosynthesis)

I.1 Site of photosynthesis (chloroplast) transverse section of dicot leaf.
I.2 Photosynthetic pigment
I.3 Absorption and light spectra
I.4 Light reactions
I.5 Dark reactions
I.5.1 C3, I.5.2 C4 and I.5.3 Cam pathways.
I.6 Photorespiration.
II- Crop plant responses to environment
 Stress of plants to various environmental conditions (dry habitat, flooded conditions
etc.)
 Applications of crop physiology in crop management (density, spacings, thinning,
pruning, and pollarding.

I- Carbon nutrition (Photosynthesis).

Carbon is the most abundant compound in nature. It is found in all organic compounds
thus often referred to as the element of organic compounds. It is available to living things from
the environment in inorganic form i.e. CO2 (atm), CO3 and HCO3 (water). The conversion of
inorganic carbon to organic compound is made possible by plants through the process of
photosynthesis.

What is photosynthesis.

This is the reduction of inorganic carbon into organic compounds. It is an endergonic

or energy consuming process in green plants and the source of energy is sunlight. It takes place
in higher plants, algae and some bacteria and it is the most important biochemical process on
earth. It is a vital process in life because it permits not only the maintenance of normal oxygen
concentration in the atmosphere but all living organisms depends on it either directly or
indirectly for energy.

The first evidence to demonstrate this process was by Joseph Priestly (1733-1804). He
identifies the ability of green leaves to produce oxygen. Using an aquatic green plant upon
illumination, gas bubbles are given off. If the illumination is stopped the production of gas
bubbles stops immediately and resumes again instantly upon illumination.

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 The primary products of photosynthesis are carbohydrate mainly starch which
accumulate as starch grains in the chloroplast. Careful analysis shows that the process only
takes place in the green parts of plants hence in cells containing chloroplast and green pigment
chlorophyll. This process can be summarised by the equation below.
CO2 + H2O = (CH2O) n + O2
During this process carbon taken up in the form of CO2 is incorporated into organic compounds
and these constitutes the starting point for all other biosynthetic processes. Hence all organic
matter in green plants may be considered as the product of photosynthesis.
Evidence to demonstrate the source of oxygen during photosynthesis.
Using radioactive isotopes, it has been shown that the oxygen produce during photosynthesis
comes from water and not from CO2. CO2 is reduced by hydrogen from water into
carbohydrates. Water is also produced during the process.
CO2 + H2O+ = (CH2O) n + H2O + O+2
To produce the hydrogen atoms, water has to be lysed or photolysis and this require large
amount of energy usually from the sun. The energy breaks the water molecules to generate
high energy electrons for the reduction of CO2.
H2O = (OH-) + (H+ + e-), OH- is very unstable and regroup to give
4(OH-) = 2H2O + O2 (given off).
The process of photosynthesis takes place in two stages
 The first stage requires light hence it called the light dependent reaction or light phase
 The second stage is the light independent reactions or dark phase.
The importance of photosynthesis
 The energy entering chloroplasts as sunlight gets stored as chemical energy in organic
compounds
 Sugar made in the chloroplasts supplies chemical energy and carbon skeletons to
synthesize the organic molecules of cells
 Plants store excess sugar as starch in structures such as roots, tubers, seeds, and fruits
 In addition to food production, photosynthesis produces the O2 in our atmosphere

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Internal structure of the leaf.

In flowering plants, the main photosynthetic organs are the leaves. The internal
structure of the leaf is different both in monocot and dicot. In dicot the structure is simple
consisting of cuticle, upper epidermis, mesophyll cells and lower epidermis while in monocot
there is complex arrangement around the vascular bundle. The leaf anatomy in the maize plant
has two rings of cells around each of the vascular bundles. The inner ring called bundle sheath
cells contain chloroplast which differ in form from those in the mesophyll cells in the outer
ring. The chloroplast is thus described as dimorphic i.e. having two forms. This type of anatomy
is called the kranz anatomy which means crown referring to the distinct rings of cells. It occurs
mostly in C4 plants such as the maize plant, sugar cane, rice etc.

Structure and functions of parts of a leaf.

a. The lower and upper epidermis. Cells are one cell layer thick, flattened and lacking
chloroplast. The upper part is covered with a waxy cuticle and the lower part contain
stomata. Each stoma is surrounded by a pair of guard cells. They function in protection,
prevent desiccation, infection and gaseous exchange.
b. The palisade mesophyll cells. They are column-shaped cells with numerous chloroplast
in a thin layer of cytoplasm. They are the main photosynthetic tissue.
c. Spongy mesophyll cells. Irregularly-shaped cells fitting together loosely to leave large
air spaces. They are also photosynthetic but with fewer chloroplast than the palisade
mesophyll cells. Gaseous exchange can occur through the large spaces through stomata.
Also store starch
d. Vascular tissues. Extensive finely branching network through the leaf. Conduct water
and mineral salt to the leaf in xylem and removes product of photosynthesis in phloem.
They also provide a supporting skeleton to the lamina.

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 How does the structure of the leaf contribute to its successful functioning?

The Chloroplast.
They are surrounded by two membranes and always contain chlorophyll and other
photosynthetic pigments located on a system of membranes. The membranes run through a
ground substance called stroma. The system consists of many flattened, fluid-filled sacs called
thylakoids which form stacks called grana at intervals with layers (lamellae) between the grana.
Each granum resembles a pile of coins and lamellae are often sheet-like. The membrane system
(thylakoid) is the site of the light dependent reactions while the stroma is site of light
independent reaction of photosynthesis. The structure is gel-like containing soluble enzymes
and other chemicals such as sugars and organic acids.

DNA

Composition of the main parts of chloroplast.

1. The envelop.
This consist of outer and inner membranes which hold content of the organelle and allow
exchange of substances with the cytosol. The membrane is differentially permeable like most
membranes. The main constituent of the envelop include.
 galacto-lipids and sulpho-lipids which are peculiar to the chloroplast envelop.
 Phosphatidylcholine.
 Carotenoids and at least 75 different proteins representing about 1-2% of all chloroplast
proteins. The membrane of the envelop does not contain chlorophyll.
The membrane performs a number of functions such as transport of substances synthesised into
the cytosol, synthesised lipids and show differential permeability.
2. Stroma. It is a colourless matrix that fills most of the volume of the chloroplast. It is a
kind of gel-like fluid that surround the thylakoids. The main constituents are.
 Ribulose bisphosphate carboxylase oxygenase (Rubisco) 50% of chloroplast proteins
and other enzymes of the Calvin cycle.
 It also contains ribosomes, starch grains, lipid droplet, DNA and RNA.

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The main function includes carbon dioxide fixation and synthesis of carbohydrates such as
starch, synthesis of fatty acid, amino acid and other proteins.
3. Thylakoids. They are flattened vesicles arranged in a membrane system. The outer
surface is in contact with the stroma and the inner surface encloses an intra-thylakoid
space called lumen. The entire surface area is about 100 times greater than the envelop.
Thylakoid may be stacked like piles of coin forming grana (granum) or they may be
unstacked forming interconnections between the grana. The unstacked thylakoids are
called stroma thylakoids or inter-grana thylakoid.
The main composition of the thylakoids includes.
5 Macromolecular complexes.
 Light harvesting complex (LHC).
 Photosystems (PSI and PSII)
 Cytochrome b6f complex
 Plastocyanin complex
 ATPsynthase complex.
Lipids such as fatty acids, phospholipids, glycolipids, sulpholipid, carotenoids, chlorophyll and
some mineral ions such as Mg, Fe, Cu etc.
Main functions of thylakoids are.
Absorption of light, electron transport, electrochemical gradient, production of ATP and
NADPH and evolution of oxygen.
Photosynthetic pigments.

The substance responsible for the absorption of sunlight in photosynthesis is the pigment
chlorophyll which is located in the chloroplast. The chloroplast of higher plants contain
chlorophyll a (blue green in chromatogram), chlorophyll b (yellow green), beta carotenoids i.e.
carotene (orange yellow) and sometimes xanthophyll (yellow). These pigment absorbs light at
various wave length.

a. Chlorophylls. This is the green pigment of plants which are the primary photoreceptors
and main energy transfer agent. They form energy trap and are primary electron donors
in photosynthesis. The chlorophyll molecule has a tetrapyloric structure called
porphyrin with the Mg atom at the centre and a long phytol chain or tail extending from
one of the four pyrrole rings. The empirical formula of the chlorophyll molecule is
C55H72O5N4Mg. the phytol chain is hydrophobic but lipophilic while the porphyrin
body is hydrophilic. There are several types of chlorophyll which differ only slightly.

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 Chlorophyll A is found universally in all photosynthetic plants, chlorophyll B which
differ from chlorophyll A in that the -CH3 attached to the third carbon atom is replaced
by a -CHO group. It is also found only in higher plants and green algae.
Chlorophyll A is the most abundant photosynthetic pigment. It exists in several forms
depending on its arrangement in the membrane.

Chlorophyll A and B show different absorption spectra. An absorption spectrum is a

measure to the extent to which a given substance absorbs light of different wavelength. It is the
function of the relationship between absorption and wavelength. Both chlorophyll A and B
show absorption maxima at the blue-violet (450nm) and orange-red regions (650nm) of the
visible spectrum and minima between 500 and 600nm. The absorption spectrum of chlorophyll
provides indirect evidence of the wavelength of light that are absorbed for the process of
photosynthesis. The absorption maxima indicate the quality of light most effective in the
process.

An action spectrum is a
graph showing the
effectiveness of the different
wavelengths of light in
stimulating the process being
investigated. It shows which
wavelength of light is most
effectively used in a specific
chemical reaction. Chlorophyll
is more efficient in using the
red-blue spectrum of light
hence the peak of reaction will
be above wavelength representing blue and red colours.

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 b. Carotenoids. Its consist of carotenes and xanthophyll. Carotenes are the major
carotenoids found in photosynthetic tissues while xanthophyll are more abundant in
nature. The most important and widespread carotene is beta carotenes, which is familiar
with the orange pigment of carrots. They are yellow, orange, red or brown pigments
that absorbs strongly in the blue-violet range. They play two important roles in
photosynthesis i.e. as accessory pigments, they absorb and transfer light energy to
chlorophyll A. They also protect chlorophyll against photo-oxidation. They are usually
masked by chlorophyll but can be seen in leaves before leaf fall. They are also found in
some flowers and fruits where the bright colours attract insects, birds and other animals
for pollination and dispersal.

Protection against photo-oxidation.

This occurs when many more chlorophyll molecules become excited than can be utilized in
photosynthesis. In the presence of oxygen, the chlorophyll molecules are subjected to oxidation
resulting to chlorosis and dead of the leaves. Chlorophyll which is activated by the absorption
of light is generally returned to its original state as a results of its participation in
photosynthesis. However, activated chlorophyll may combine with molecules of oxygen to
form a complex that can lead to photo-oxidation.

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 UV

Chl

Chl+ photosynthesis

Chl+ O2
Complex.

The excited Chl+ oxygen complex can be inactivated by carotenoids. It is thought that
carotenoids protect chlorophyll photo-oxidation by acting as preferred substrate in
photosynthesized oxidation. They are oxidised through light catalysed formation of epoxides
across double bonds and are then reduced by a dark enzyme catalysed reaction.
Light
carotenoids
Epoxy-carotenoids
catalysed
Carotenoids may also absorb light energy and somehow divert it from chlorophyll through heat
dissipation. Thus carotenoids provide a channel to get rid of excess energy absorbed by
chlorophyll.

The light reaction of photosynthesis.

In 1937 Robert Hill showed that isolated chloroplast can evolve oxygen and acquire reducing
power when illuminated even in the absence of CO2. He then concluded that water had been
split into hydrogen and oxygen. This reaction was called the Hill reaction and is one of the
main events in the light stage of photosynthesis. It has also been shown that isolate chloroplast
is capable of producing ATP as well. Hence in the light phase hydrogen atoms are produced
for the reduction of CO2 in the dark phase while ATP is the source of energy for the subsequent
synthesis of carbohydrate.

Light absorption by chlorophyll.

Light energy is absorbed by pigment molecules of chlorophyll which causes excitation. The
potential energy of the absorbing molecule is increased by the displacement of one or more
electrons to an outer orbital. After excitation the energy can either be dissipated by fluorescence
and lost as heat or transferred to another molecule such as adjacent chlorophyll molecule by
resonance. The second molecule becomes excited while the first return to ground state. Certain
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special chlorophyll molecules called reaction centre chlorophyll when excited loose electrons
to a neighbouring electron acceptor.
Light energy
Chl
Chl+ + e-
Reduced form
Oxidised form

This initial separation of charges is the basic reaction in photosynthesis. Each reaction centre
is associated with the light harvesting of the other pigment molecules that can provide energy
to it. Such a complex is often called a photosynthetic unit and comprised of a reaction centred
chlorophyll, the light harvesting pigment chlorophyll molecule which are the accessory
pigment and the electron transport chain. The photosynthetic unit is the smallest group of
collaborating pigment molecules necessary to effect a photochemical reaction. The tight
arrangement of chlorophyll molecules in the grana allows for excellent energy transfer by
resonance. The amount of light energy absorbed by a single antenna chlorophyll move from
one molecule to another until it is dissipated as heat, fluorescence or formation of ATP and
reducing power (NADPH).

The Emersion effect.

When Emersion and co-workers exposed green plants to different wavelength of light. They
observed that at wavelengths of greater than 680 nm, the efficiency of photosynthesis decreased
abruptly. When plants were exposed to short-wavelength light of less than 660nm, the
efficiency of photosynthesis also decreases. Green plants were then exposed to both short and
long wavelengths at the same time, the efficiency of photosynthesis increased greatly. They
concluded that there must be two photosystems involve in photosynthesis one driven by short
wavelength light and the other driven by long wavelength of light (PSI and PSII). They work
together to enhance efficiency and convert the light energy to forms that can be absorbed by
plants. Light excites the chlorophyll molecules at the reaction centre and cause an increase in
energy. As the molecule becomes less excited, its energy is transported through a chain of
electron carriers to the next photosystem which does much the same thing and produces energy
carrying organic molecules.

Photosystems.

The discovery of the emersion effect and subsequent research showed that photosynthesis
require the interaction of two distinct groups of pigments called photosystems (PSI and PSII).

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PSI has high chl a / chl b ratio and absorbs light of wavelength greater than 680nm with
absorption peak at 700nm. The reaction centre chlorophyll molecule is at P700.

PSII has low chl a /chl b ratio and absorbs light of wavelength less than 680nm with absorption
peak at 680nm. The reaction centre chlorophyll molecule is at P680.

Each photosystem is made up of a light harvesting complex (LHC), a reaction centre with
special chl a molecules P700 in PSI and P680 in PSII. This arrangement is based on the fact that
the majority of chloroplast pigment does not take part in photosynthesis directly but absorbs
light energy and transfer it to the reaction centre where the special chl a molecule can ionise
and initiate the movement of electrons in a redox cycle. Carotenoids, chl b and chl b are
arranged in a chain of energy transport where energy is transferred by resonance because the
inter-molecular distance and orientation is very specific. The path taken by energy transfer in
the LHC is hap-hazard until it gets to the reaction centre. In the reaction centre special chl a
molecule ionises by loss of an electron thus initiate the electron transport chain.

Electron transport and photophosphorylation.

The energy absorbed by a pigment molecule may lead to charge separation i.e. production of
electrons. The primary flow of electrons within a given granum of thylakoid may be initiated
almost simultaneously for each photosystem. Through integrated reactions and the lysis of
water which provide the necessary electrons flow to produce ATP and NADPH.

Non-cyclic electron flow or non-cyclic photophosphorylation (The Z scheme).

When light of appropriate wavelength falls on PSI i.e. P700 is excited and electrons the move
spontaneously to a primary electron acceptor through an electron transport chain.

Q ADP + Pi A (FeS) Fd NADP+

ATP NADPH

e-
4H+
P700 (PSI)
P680 (PSII)

2H2O = 4H++ 4e-+ O2

The primary acceptor of electron is A (FeS protein). Electrons are transferred to ferredoxin
(Fd) and then to NADP+. NADP++ 2H+ + 2e- = NADPH + H+(reducing power). The transfer
of electrons creates a hole in PSI. this hole or deficit is filled by the excitation of P680 in PSII
which lead to the release of electrons and their transfer to PSI through a system of electron
carriers. This include Quinone(Q), plastoguinone (PQ), cytochrome f (Cyt f) and cytochrome
b6 (Cyt b6) and plastocyanin (Pc). The hole created in PSII is filled by electrons from water.

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Electrons transported between the two photosystems is from a higher energy level to a lower
energy level. This electrochemical gradient is used in the formation of ATP
(photophosphorylation).
Cyclic electron flow or photophosphorylation

When the chloroplasts are illuminated with light of wavelength above 680nm, non-cyclic
photophosphorylation can be excluded. Under this condition, only PSI is activated and
electrons are not removed from water and CO2 assimilation is consequently retarded. As such
NADP+ is no longer available as an electron acceptor. Wavelength above 680 activate PSI
causing electrons to flow from P700 to A (FeS). With no NADP+ to receive the electrons they
are passed back to P700 through cytochrome b6, cytochrome f and plastocyanin. Cyclic flow
is used to demonstrate the cycling of electrons from the donor (excited PSI) to an acceptor A
(FeS) and back to P700 in PSI with limited generation of ATP. It is thought that it provides the
extra ATP needed for CO2 fixation.

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Light independent reactions of photosynthesis (dark phase).

This phase is essentially involving in CO2 fixation and its subsequent reduction to
carbohydrate. This is an endergonic reaction and energy is supplied by ATP formed in the light
phase. Calvin and Benson (1946-1945) working with a radioactive isotope of carbon (14C)
succeeded in mapping and identifying the intermediates involved in the assimilation of CO2 in
photosynthesis.

The Calvin cycle or C3 carboxylation cycle.

The most important stages in carbodioxide reduction in the C3 cycle are.
1. Carbodioxide fixation or carboxylation.
CO2 is accepted by ribulose-1,5-biphosphate (RubP) in the presence of water to form an
unstable intermediate which undergo cleavage enzymatically to form two molecules of
phosphoglyceric acid (3-PGA).
CO2 + CH2O(P)CO-CHOH-CHOH-CH2O(P) +H2O 2CH2O(P)-CHOH-COOH
RubPcarboxylase.
The enzyme that catalyse this reaction is ribulose bi-phosphate carboxylase which is probably
the most abundant enzyme in photosynthetic tissues. The 2 molecules of 3-PGA are converted
to 1,3-diphosphoglyceric acid using two molecules of ATP from the light reaction.
phosphoglyceric acid (PGA) is the first stable product of dark reaction of photosynthesis and
since it is a 3 carbon compound, this cycle is known as C3 cycle.
3PGA Kinase. CH20(P)-CHOH-COOP
CHO(P)-CHOH-COOH
ATP ADP

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 2. Reduction of PGA to PGAL (phosphoglyceraldehyde).

This is the most crucial phase in the cycle. Six molecules of 1, 3 diphosphoglyceric acid are
reduced with the use of 6 molecules of NADPH2 (produced in light reaction) to form 6
molecules of glyceraldehyde 3-phosphate. This reaction is catalysed by glyceraldehyde 3-
phosphate dehydrogenase.
Glyceraldehyde 3-P dehydrogenase CH2O(P)-CHOH-CHO
CH2O(P)-CHOH-COO(P)
NADPH NADP+ + Pi

Glyceraldehyde 3-phosphate is very important in the cycle. It may be transported out of the
chloroplast and converted into various hexoses (glucose, fructose, sucrose etc.), converted to
starch and stored in the chloroplast or move to metabolic pool.

3. Regeneration of RubP.

The regenerative phase is called as pentose phosphate pathway. For the 6 molecules of 3-PGAL
produced, one molecule enters the metabolic pool as a net gain in energy and substrate to the
metabolic system. The remaining 5 molecules are inter-converted through a series of reactions
into various phosphorylated sugars. A 3 molecules of ribulose 5-phosphate is synthesised
which then react with ATP from the light reaction to form ribulose 1,5- bisphosphate. Ribulose
1,5-bisphosphate ensures the continuity of the cycle.

4. Product synthesis.

This include sugars, lipids, fatty acids, amino acids and other organic acids. These are
synthesised based on the light condition and CO2 tension.

Schematic diagram of light reaction and Calvin cycle

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 The incorporation of one CO2 molecule by RubP give rise to 2PGA and finally 2, 3-
carbon sugars. This 3-carbon sugar is used for the synthesis of carbohydrates and regeneration
of RubP. Ths the incorporation of 3 CO2 molecule will give rise to six 3-carbon sugar
molecules, five of will be used to regenerate RubP for continuity of the cycle.

NB. The first stable product formed after CO2 fixation through the Calvin/Benson cycle is a
3-carbon compound (PGAL) thus it is refered to as the C3 pathway and plants which fix CO2
exclusively through this pathway are called C3 plant. Though a vast majority of plants fix
CO2 through other pathways.

C4 and CAM Pathways.

Carbondioxide fixation in C4 plants (the hatch-slack pathway).
In some plants especially tropically grasses such as maize plant, sugar cane, sorghum,
millet etc. the initial fixation of radioactive carbon (14C) after very short period of
photosynthesis in the presence of radioactive CO2 is predominantly malic and aspartic acid.
Approximately 93% of the radioactive carbon is incorporated into these acid within a short
period of time. Radioactivity appears in PGAL only after a relatively longer period. Also
ribulose 1,5-bisphosphate carboxylase, the enzyme that catalyse the carboxylation is neither
active or concentrated in the mesophyll cells of these plants. Instead the enzyme phosphoenol
pyruvate carboxylase (pepcarboxylase) which catalyse carboxylation into oxaloacetic acid is
found in relatively higher concentration.

Hatch and Slack (1966-1967) were able to detect the relatively unstable radioactive
carbon (14C) in oxaloacetic acid as the first product of carboxylation of phosho-enol pyruvate.
Because this product a 4-carbon compound, plants exhibiting this pathway are are called C4
plants. CO2 is incorporated to PEP in the presence of enzyme pepcarboxylase to form the first
product which is oxaloacetic acid which is later converted to malic acid and aspartic acid.

COOHCHOHCH2COOH
PEPcarboxylase
PEP carboxylase
CH2=CO(P)COOH COOHC=OCH2COOH Malic acid

Oxaloacetic acid
COOHCHNH2CH2COOH
CO2
Aspartic acid

The leaf anatomy in C4 plants is very perculiar. The leaf anatomy in the maize plant has two
rings of cells around each of the vascular bundles. The inner ring called bundle sheath cells
contain chloroplast which differ in form from those in the mesophyll cells in the outer ring.
The chloroplast is thus described as dimorphic i.e. having two forms. This type of anatomy is
called the kranz anatomy. It occurs mostly in C4 plants such as the maize plant, sugar cane,

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rice etc. The mesophyll cells have high pepcarboxylase activity while the bundle sheath cells
have high RubPcarboxylase activity and the other enzymes of C3 cycle. Thus the leaves of C4
plants are compartmentalised and show division of labour. There is CO2 fixation into C4 acid
in chloroplast of mesophyll cells and subsequent formation of phosphorylated sugar and starch
in the chloroplast of the bundle sheath cells. Oxaloacetate is transported from mesophyll cells
to bundle sheath chloroplast where decarboxylation takes place. The CO2 liberated is then
incorporated into the C3 cycle.

NB.

 C4 plants are more productive because PEPcarboxylase has a high affinity for
carbodioxide. They absorb CO2 strongly from a much lower concentration than C3
plants. They have limited phosphorespiration activity because even if it happens, the
CO2 liberated is recaptured and fixed by mesophyll cells.
 Localisation of C3 enzymes in the bundle sheath cell provide a favourable and efficient
carbohydrate metabolism i.e. a port for the loading and transport of sucrose in the
phloem.
 C4 plants also use more light energy in photosynthesis thus are limited to the tropic
where there is excess light.

CAM Plants (Crassulacean Acid Metabolism).

CAM plants are usually succulents and they grow under extremely xeric (dry) conditions.
They are adapted to photosynthesis and survival under adverse xeric (dry) conditions.
Examples are Cactus, Orchid, Pineapple, Kalanchoe etc. The mesophyll cells have larger
number of chloroplasts and the vascular bundles are not surrounded by well-defined bundle

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 sheath cells. In these plants, the stomata remain open during night and closed during the
day. They fix CO2 with accompanying acidification. The acid content of the plant increase
during the night and decreases during the day. They fix CO2 into malic acid like C4 plants
but lack the kranz anatomy. Phospoenolpyruvate is incorporated into carbondioxide catalyse
by enzyme pepcarboxylase into malic acid which is stored in the vacuole. During the day
when the stomata are closed, the malic acid undergo decarboxylation releasing CO2 for
carbohydrate formation through the Calvin cycle in the mesophyll cells. This is an inefficient
pathway due to the fact that the carboxylation enzyme is located in the cytoplasm whereas
the malic acid is stored in the vacuole. Hence no metabolism will occur until the

concentration of malic acid becomes high so that no more can enter.

PEP + CO2 + H2O Oxaloacetate + Pi

NADPH NADP+
Oxaloacetate Malic acid Acidification
Malic dehydrogenase

NADP+ NADPH
Malic acid Pyruvate + CO2 Deacidification
Malic enzyme

Photorespiration.

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 The excessive respiration that takes place in green cells in the presence of light. This
was first demonstrated by Warburg (1883-1970) that plant consume oxygen at a higher rate
when they were illuminated. He also found out that this increased in oxygen consumption
inhibited photosynthesis. Several suggestions were made to explain this effects. That
temperature and oxygen concentration play an important role. Photorespiration is very high
when the temperature is between 25 and 30 ºC. The rate of photorespiration increases with
the increase in the concentration of oxygen. Three cell organelles namely chloroplast,
peroxisome and mitochondria are involved in the photorespiration. This kind of respiration
is seen in plants like cotton, capsicum, peas, tomato, soybean, wheat, etc. (mostly C3 plants)
and it is absent in grasses (C4 plants). Further investigation in the 70’s showed the rate of
respiration (O2 consumption and CO2 liberation) from the leaves of C3 plants was two times
greater in light than in the dark. It occurs in the mesophyll cells of C3 plants at high O2
concentration and temperature though has been observed in the bundle sheath cells of C4
plants.

This was explained by Ogren and Bowes (1971) that the site of action of O2 was
ribulose 1, 5-bisphosphate carboxylase oxygenase (Rubisco) and the substrate is RubP. In
the presence of O2, the enzyme operates as an oxygenase catalysing the oxidation of RubP
to phosphoglycolate. PGA
CO2 Rubisco COOHCHOHCH2O(P) C3 Cycle

CH2O(P)COCHOHCHOHCH2O(P)

CH2O(P)COOH C2 cycle
Rubisco
O2 phosphoglycolate CH2OHCOOH
Glycolic acid
Oxygen thus compete with CO2 for RubP causing a reduced rate of fixation of
phosphorylated sugar in the chloroplast. Phosphoglycolate is then converted to glycolic acid
which is transferred to peroxisomes where the glycolic acid is oxidised to glyoxylic acid and
hydrogen peroxide. The glyoxylic acid is converted into glycine which is an amino acid.
Two molecules of glycine condense to form serine and liberate carbon dioxide. Serine is
metabolised in the mitochondria to produce carbohydrate or incorporated into proteins or
recycled in the chloroplast.

NB. Since RubP carboxylase acts as a carboxylase as well as an oxygenase, it is usually

referred as Rubisco (ribulose bisphosphate carboxylase oxygenase. Its activities as a
carboxylase or an oxygenase depends on CO2 concentration of the medium.

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The importance of photorespiration.

 Help to maintain CO2 concentration in the leaves when the stomata are closed may
be due to water stress.
 Might serve to dissipate unwanted energy when light intensity is too high and CO2
in short supply
 Excess oxygen is toxic to plants thus through formation of reactive radicals H2O2, it
might help to maintain the level of O2 in the chloroplast below toxic threshold
especially under stress condition when the stomata are closed.
 CO2 from this process might help to keep the C3 cycle turning and to maintain the
level of intermediates
 Provide enough CO2 to maintain in the active state and prompt photosynthesis upon
opening of stomata.
 It provides a mechanism for intracellular transport and interconversion of
carbohydrate to nitrogen compounds like from phosphoglycolate to glycine, serine
and to PGA.

Comparison of the plants of C3 and C4 cycle

C3 Plant C4 Plant

1. Only C3 cycle is found Both C4 and C3 cycles are found.

2. The efficiency of CO2 absorption at low The efficiency of CO2 absorption at low concentration
concentration is far less and hence, they are less is quite high and hence, they are more efficient plants.
efficient.
3. The CO2 acceptor is Ribulose-1, 5diphosphate. The CO2 acceptor is phosphoenolpyruvate.

4. The first stable product is phosphoglyceric acid Oxaloacetate (OAA) is the first stable product.
(PGA).
5. Plants show one type of chloroplast Plants show dimorphic type of chloroplast. The
(monomorphic type). chloroplast of bundle sheath is different from that of

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 mesophyll cells (dimorphic type). The chloroplasts in
bundle sheath cell are centripetally

arranged and lack grana. Leaves show Kranz type of

anatomy.

6. In each chloroplast, two pigment systems In the chloroplasts of bundle sheath cells, the
(Photosystem I and II) are present. photosystem II is absent. Therefore, these are
dependent on mesophyll chloroplasts for the supply of
NADPH + H+.
7. The Calvin cycle enzymes are present in Calvin cycle enzymes are absent in mesophyll
mesophyll chloroplast. Thus, the Calvin cycle chloroplasts. The cycle occurs only in the chloroplasts
occurs. of bundle sheath cells.
8. The CO2 compensation point is 50-150 ppm CO2. The CO2 compensation point is 0-10 ppm CO2.
9. Photorespiration is present only to a slight degree or
Photorespiration is present and easily detectable.
absent.
10. The CO2 concentration inside leaf remains high The CO2 concentration inside the leaf remains low
(about 200 ppm). (about 100 ppm).
13 12
11. The C/ C ratio in C-containing compounds
The ratio is relatively high, i.e. C4 plants are more
remains relatively low (both 13CO2 and 12CO2 are
enriched with 13C than C3 plants.
present in air).
12. Net rate of photosynthesis in full sunlight (10,000 It is 40-80 mg. of CO2 per dm2 of leaf area per hour.
– 12,000 ft. c.) is 15-25 mg. of CO2 per dm2 of That is, photosynthetic rate is quite high. The plants
leaf area per hour. are efficient.
13. The light saturation intensity reaches in the range
It is difficult to reach saturation even in full sunlight.
of 1000-4000 ft. c.
14. Bundle sheath cells are unspecialized. The bundle sheath cells are highly developed with
unusual construction of organelles.
15. The optimum temperature for the process is 10- In these plants, it is 30-45°C and hence, they are warm
25°C. climate plants. At this temperature, the rate of
photosynthesis is double than that is in C3 plants.
16. 18 ATPs are required to synthesize one glucose 30 ATPs are required to synthesize one glucose
molecule. molecule.

Factors affecting photosynthesis

The rate of photosynthesis is an important factor in crop production since it affects yields. An
understanding of those factors affecting the rate is therefore likely to lead to an improvement
in crop management.
Limiting factors.
The rate of any biochemical process just like photosynthesis involves a series of reactions and
will be limited theoretically by the slowest reaction in the series. Example in photosynthesis,
the light independent reactions are dependent on the light dependent reactions for reduced
NADP and ATP. At low light intensity, the rate at which these are produced is too slow to
allow the light-independent reactions to proceed at maximum rate thus light becomes a limiting
factor.

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The principle of limiting factor state as follows-when a chemical reaction process is affected
by that factor which is nearest it’s minimum value, it is that factor which directly affects a
process if it’s quality is changed.

Exercise. Study the graphs below and answer the questions that follows.
1. In fig (1) i. what is the limiting factor in region A? ii. what is represented by the curve
at B and C? iii. what does point D represent on the curve? iv. What does point E
represent on the curve?
2. In fig (2) i- what does the point X, Y and Z represent on the three curves?

Rate of photosynthesis

(fig 1) Light intensity (fig 2)

I. External factors
1. Light
It is the most important factor of photosynthesis. Any kind of artificial light such as electric
light can induce photosynthesis. Out of the total solar energy, only 1-2 % is used for
photosynthesis and the rest is used for other metabolic activities. The effect of light on
photosynthesis can be studied under three categories.
a. Light intensity
Wolkoff (1966) found that the arte of photosynthesis is directly proportional to light
intensity. But the extremely high light intensities do not favour higher photosynthetic rates.
The high light intensity which fails to accelerate photosynthesis is called light saturation
intensity. Of the light falling on a leaf, about 80 per cent is absorbed, 10 per cent is reflected
and 10 % is transmitted. The rate of photosynthesis is greater in intense light than in diffused
light. The plants are grouped into two types on the basis of light requirement. i. Heliophytes
(Sun plants) ii. Sciophytes (Shade plants)
At a specific light intensity, the amount of CO2 used in photosynthesis and the amount of
CO2 released in respiration are volumetrically equal. This specific light intensity is known
as light compensation point.
At very high light intensity, beyond a certain point, the photosynthetic cells exhibit photo
oxidation. This phenomenon is called solarisation and a result of this, inactivation of

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chlorophyll molecules, bleaching of chlorophyll molecules and even inactivation of some
enzymes take place resulting in the destruction of whole photosynthetic apparatus. In
general, low light intensity favours stomatal closure and in turn reduced rate of
photosynthesis.
b. Light quality (wavelength)
Photosynthesis occurs only in the visible part of the light spectrum i.e., between 400 and
700 nm. The maximum rate of photosynthesis occurs at red light followed by blue light.
The green light has minimum effect and photosynthesis cannot take place either in the
infrared or in the ultraviolet light.
c. Light duration
In general, tropical plants get 10-12 hours of light per day and this longer period of light
favours photosynthesis.
2. Carbon dioxide
CO2 is one of the raw materials required for photosynthesis. If the CO2 concentration is
increased at optimum temperature and light intensity, the rate of photosynthesis increases.
But, it is also reported that very high concentration of CO2 is toxic to plants inhibiting
photosynthesis.
3. Temperature
The rate of photosynthesis increases by increase in temperature up to 40 ºC and after this,
there is reduction in photosynthesis. High temperature results in the denaturation of enzymes
and thus, the dark reaction is affected. The temperature requirement for optimum
photosynthesis varies with the plant species. For example, photosynthesis stops in many
plants at 0 ºC but in some conifers, it can occur even at -35 ºC. Similarly, photosynthesis
stops beyond 40-50 ºC in certain plants; but certain bacteria and blue green algae can perform
photosynthesis even at 70 ºC.
4. Water
Water has indirect effect on the rate of photosynthesis although it is one of the raw materials
for the process. The amount of water utilized in photosynthesis is quite small and even less
than 1 per cent of the water absorbed by a plant. Water rarely acts as a limiting factor for
photosynthesis. During water scarcity, the cells become flaccid and the rate of
photosynthesis might go down.
5. Oxygen
Oxygen is a bi-product of photosynthesis and an increase in the O2 concentration in many
plants results in a decrease in the rate of photosynthesis. The phenomenon of inhibition of

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photosynthesis by O2 was first discovered by Warburg (1920) in green alga Chlorella and
this effect is known as Warburg’s effect. This is commonly observed in C3 plants.
In plants, there is a close relationship between Warburg’s effect and photorespiration. The
substrate of photorespiration is glycolate and it is synthesized from some intermediates of
Calvin’s cycle. In plants that show Warburg’s effect, increased O2 concentration result in
diversion of these intermediates of Calvin cycle into the synthesis of glycolate, thereby
showing higher rate of photorespiration and lower photosynthetic productivity.
6. Mineral elements
The elements like Mg. Fe, Cu, Cl, Mn, P etc are involved in the key reactions of
photosynthesis and hence, the deficiency of any of these nutrients caused reduction in
photosynthesis.
7. Chlorophyll content
It is very much essential to tarp the light energy. In 1929, Emerson found direct relationship
between the chlorophyll content and rate of photosynthesis. In general, the chlorophyll
sufficient plants are green in colour showing efficient photosynthesis. The chlorotic leaves
due to irregular synthesis of chlorophyll or breakdown of chlorophyll pigment exhibit
inefficient photosynthesis.
8. Leaf
The leaf characters such as leaf size, chlorophyll content, number of stomata. Leaf
orientation and leaf age are some of the factors that are responsible for photosynthesis. The
maximum photosynthetic activity is usually seen in the physiologically functional and full
size leaves (usually third/fourth leaf from the tip of the shoot system).
9. Carbohydrates
If the accumulated carbohydrates are not translocated, the photosynthetic rate is reduced and
respiration is increased. Sugar is converted into starch and gets accumulated in the
chloroplasts. This reduces the effective surface in the chloroplast and the rate of
photosynthesis is decreased.
10. Phytohormones
Photosynthesis may be regulated by plant hormone system. Gibberellic acid and cytokinin
increase the carboxylating activity and photosynthetic rates. It also reported that kinetin at
3µm concentration causes 12 per cent increase in photosynthesis within one hour of the
treatment.

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 Tutorials.
1. What is the significance of photosynthesis to man?
2. Why are C4 plants more productive than C3 plants
3. What is the basis of classifying plants into C3, C4 and CAM?
4. i-Give the different reactions that take place in PSII (equations only)
ii- What is the composition of this photosystem
iii How does it relate with photosystem I.
5. i-What are C4 plants
ii-Do they possess the enzyme RuBPcarboxylase? Explain
iii- Why is osmosis more pronounce in C3 plants than C4 plants
6. Explain the following as used in photosynthesis
i-Emersion effect, ii-limiting factors, iii-photorespiration
7. What has been the importance of radioisotopes in photo-research
8. a-How is photorespiration different from oxidative phosphorylation
b-How does both interfere in photosynthesis
c-Why is it that after 3hrs of darkness CO2 fixation is impossible.
9. a-Draw a graph of showing variation in photosynthesis with light intensity in a plant
shaded and a plant exposed to the sun
b-Explain fully the shape of your graph in not more than 5lines.
10. In other to have a photochemical effect in a plant, light must be absorbed by
pigments.

a. State the various photosynthetic pigments

b. How can you justify the participation of these pigments in photosynthesis.

c. Give the significance of the participation of several pigments in photosynthesis

d. What is the role of each pigment in photosynthesis.

11. Write down the location of the following processes:

a. Light reactions of photosynthesis
b. Dark reactions of photosynthesis
c. Electron transport chain.

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