F214 Module 3: Photosynthesis

4.3.1 Photosynthesis
(a) define the terms autotroph and heterotroph
Autotrophs - organisms that use light energy or chemical energy and inorganic molecules (carbon dioxide and water)
to synthesise complex organic molecules.
Heterotrophs - organisms that ingest or digest complex organic molecules, releasing the chemical potential energy
stored in them.
Phototrophs - organisms that uses energy from sunlight to synthesise organic compounds for nutrition.
Chemoautotrophs – organisms which synthesise complex organic molecules using light energy derived from
exergonic chemical reactions.

(b) state that light energy is used during photosynthesis to produce complex organic molecules
Photosynthesis is the process whereby light energy from the Sun is transformed into chemical energy and used to
synthesis large/complex organic molecules from inorganic substances.

(c) explain how respiration in plants and animals depends upon the products of photosynthesis
Both photoautotrophs and heterotrophs can release the chemical potential energy in complex organic molecules
(made during photosynthesis) - this is respiration. They can also use the oxygen for aerobic respiration.

The equation below summarises the process of photosynthesis: 6CO2 + 6H2O (+ light energy) → C6H12O6 + 6O2

Once the Earth's atmosphere contained free oxygen, organisms evolved that could use the oxygen for aerobic
respiration. This releases carbon dioxide back into the atmosphere and produces water.
C6H12O6 + 6O2 → 6CO2 + 6H2O (+ energy, some as ATP)

(d) state that in plants photosynthesis is a two-stage process taking place in chloroplasts
In plants photosynthesis is a two-stage process (light-dependent and light-independent) taking place in
chloroplasts.

(e) explain, with the aid of diagrams and electron micrographs, how the structure of chloroplasts enables them to
carry out their functions
 Most are disc-shaped.
 Between 2-10µm long.
 Intermembrane space is
10-20nm wide.
 Outer membrane is
permeable to small ions.
 Inner membrane is less
permeable and has
transport proteins
embedded in it.

 The inner membrane, with its transport proteins, can control entry and exit of substances between the
cytoplasm and the stroma.
 The many grana, consisting of stacks of up to 100 thylakoid membranes, provide a large surface area for the
photosynthetic pigments, electron carriers and ATP synthase enzymes (light-dependent reaction).
 Photosynthetic pigments are arranged into photosystems, which allows maximum absorption of light energy.
Proteins embedded in the grana hold the photosystems in place.
 The fluid-filled stroma contains the enzymes needed to catalyse the reactions of the light-independent stage of
photosynthesis.
 The grana are surrounded by the stroma so the products of the light-dependent reactions, which is needed for
the light-dependent reaction, can readily pass into the stroma.
 Chloroplasts can make some of the proteins they need for photosynthesis, using genetic instructions in the
chloroplast DNA, and the chloroplast ribosomes to assemble the proteins.
 F214 Module 3: Photosynthesis
(f) define the term photosynthetic pigment
Photosynthetic pigments - molecules that absorb light energy. Each pigment absorbs a range of wavelengths in the
visible region and has its own distinct peak of absorption. Other wavelengths are reflected. They appear to us as the
colour of the light wavelengths that they are reflecting.

(g) explain the importance of photosynthetic pigments in the photosynthesis

There are many different pigments that act together, to capture as
much light energy as possible. They are in thylakoid membranes,
arranged in funnel-shaped structures called photosystems, held in
place by proteins.

Photosystem - a funnel-shaped light-harvesting cluster of

photosynthetic pigments, held in place in the thylakoid
membrane of a chloroplast. The primary pigment reaction centre
is a molecule of chlorophyll a. The accessory pigments consist of
molecules of chlorophyll b and carotenoids.

How is light harvested in the chloroplast membranes?

 (Primary and accessory) pigments form photosystem/antenna complex.
 Photon/light energy absorbed by pigment molecules.
 Electron becomes excited and moves to a higher energy level.
 Energy is passed from one pigment to another.
 Energy is passed to reaction centre/chlorophyll a/ primary pigment.
 A range of accessory pigments allow range of wavelengths to be absorbed.

Primary Pigments Accessory Pigments

 Found in the reaction centre.  Found in the antenna complex.
 Chlorophyll a absorbs blue and red light, of  Chlorophyll b absorbs blue and red light, of
wavelength of around 450nm. wavelengths around 500-640nm.
o P680 - found in photosystem II and its peak of  Carotenoids absorb blue light but reflect yellow and
absorption is light at a wavelength of 680nm. orange light.
o P700 - found in photosytem I and its peak of o Carotene - orange.
absorption is light at a wavelength of 700nm. o Xanthophyll - yellow.

(h) state that the light-dependent stage takes place in thylakoid membranes and that the light-independent stage
takes place in the stroma
The light-dependent stage of photosynthesis takes place on the thylakoid membranes of the chloroplasts.
The light-independent stage of photosynthesis takes place in the stroma of chloroplasts.

(i) outline how light energy is converted to chemical energy (ATP and reduced NADP) in the light-dependent stage
(reference should be made to cyclic and non-cyclic photophosphorylation, but no biochemical detail is required)
Non-cyclic Photophosphorylation:
1. Light energy hits photosystem I and II.
2. The electrons become excited in chlorophyll a and moves to a higher energy level.
3. The electrons are accepted by electron acceptors and the move along the electron transport chain.
 The electrons lost in photosystem II are replaced by the electrons made in the photolysis of water (forms
hydrogen ions and oxygen).
 The electrons lost in photosystem I are replaced by the electrons from photosystem II.
4. Protons are pumped across the thylakoid membrane by the flow of the electrons from the stroma to the
thylakoid space, down the proton gradient.
5. The hydrogen ions made in the photolysis of water combines with electrons to form hydrogen atoms. The NADP
combines with hydrogen to form reduced NADP.
6. As the protons move across the thylakoid membrane by chemiosmosis through ATP synthase, causing
photophosphorylation producing ATP.
 F214 Module 3: Photosynthesis

Cyclic Photophosphorylation:
1. Only photosystem I is used (P700). The
excited electrons pass to an electron
acceptor and back to the chlorophyll
molecule from which they were lost.
2. There is no photolysis of water and no
generation of reduced NADP, but small
amounts of ATP are made. This may be
used in the light-independent reaction
of photosynthesis or it may be used in
guard cells (their chloroplasts only
contain only photosystem I) to bring in
potassium ions, lowering the water
potential and causing water to follow
by osmosis. This causes the guard cells
to swell and opens the stomata.

(j) explain the role of water in the light- dependent stage

Photosystem II contains an enzyme that, in the presence of light, can split water into H+ ions (protons), electrons
and oxygen. This splitting of water is called photolysis.
2H20  4H+ + 4e- +O2
Water is a source of:
 Hydrogen ions – used in chemiosmosis to produce ATP. These protons are then accepted by a coenzyme NADP,
which becomes reduced NADP, to be used into the light-independent stage to reduce carbon dioxide and
produce organic molecules.
 Electrons – replace those lost by the oxidised chlorophyll and to reduce chlorophyll a.
 Oxygen – used by the plant in aerobic respiration but much of it diffuses out of the leaves, through stomata,
into the air.
 F214 Module 3: Photosynthesis
(k) outline how the products of the light-dependent stage are used in the light-independent stage (Calvin cycle) to
produce triose phosphate (TP) (reference should be made to ribulose bisphosphate (RuBP), ribulose bisphosphate
carboxylase (rubisco) and glycerate 3-phosphate (GP), but no other biochemical detail is required)
1. In the stroma, carbon dioxide combines with ribulose bisphosphate (5C), catalysed by the enzymerubisco. RuBP
becomes carboxylated to make a 6C compound.
2. The 6C compound is unstable so breaks down into 2 molecules of glycerate 3-phosphate – carbon fixation.
3. The products from the light-dependent stage are used to reduce and phosphorylate GP into another
3Ccompound, triose phosphate.
4. The TP is used to regenerate RuBP using ATP, so that the cycle can start again.

(l) explain the role of carbon dioxide in the light-independent stage (Calvin cycle)
Carbon dioxide is the source of carbon for the production of all large organic molecules. These molecules are used
as structures, or act as energy stores or sources, for all the (carbon-based) life forms on this planet.

(m) state that TP can be used to make carbohydrates, lipids and amino acids
GP can be made into: TP can be made into:
 Amino acids  Glycerol (+ fatty acids)  Lipids
 Fatty acids (+ glycerol)  Lipids  Hexose sugars (e.g. glucose) 
o Fructose (+ glucose  sucrose)
o Polysaccharides:
 Starch
 Cellulose

(n) state that most TP is recycled to RuBP

Most triose phosphate is recycled to ribulosebisphosphate. 5 out of 6 molecules of TP (3C) are recycled by
phosphorylation, using ATP from the light-dependent reaction, to 3 molecules of RuBP (5C).
 F214 Module 3: Photosynthesis
(o) describe the effect on the rate of photosynthesis, and on levels of GP, RuBP and TP, of changing carbon dioxide
concentration, light intensity and temperature
Carbon Dioxide Light Intensity Temperature
Concentration
 More CO2 = more CO2fixation =  Light intensity – a measure of  Increase temperature = little effect
more GP = more TP = more how much energy is associated upon the rate of the light-
regeneration of RuBP. with the light. dependent reaction (no enzymes
 The number of stomata that  Distance from the source is involved), however it will alter the
open to allow gaseous doubled = the light intensity is rate of light-independent reaction.
exchange leads to increased quartered – inverse square law:  Temperature rises above 25oC =
transpiration = plant wilts if oxygenase activity of rubisco
water uptake from the soil increases more than its carboxylase
cannot exceed water loss by  Increase in light intensity = activity increases –
transpiration = stress response o More light energy available photorespiration exceeds
= release of a plant growth to excite more electrons. photosynthesis.
regulator (abscisic acid) and o More ATP and more  ATP and reduced NADP from the
stomata close = reduce reduced NADP produced, light-dependent reaction are
CO2uptake and reduce the rate which can be used in the dissipated and wasted = reduces
of photosynthesis. light-independent stage as overall rate of photosynthesis.
sources of hydrogen and  High temperature = damage
energy. proteins.
 Decrease in light intensity =  Increased temperatures = increase
o GP cannot be changed to in water loss from leaves by
TP, so GP will accumulate transpiration = closure of stomata
and levels of TP will fall. = reduction in rate of
o Lower the amount of RuBP, photosynthesis.
reducing CO2fixation and
the formation of more GP.

(p) discuss limiting factors of photosynthesis with reference to carbon dioxide concentration, light intensity and
temperature
Limiting factor - a factor that prevents a process from increasing any further at its lowest value.
CO2 Concentration Light Intensity Temperature
 Increasing the concentration of  The rate of photosynthesis is  Between the temperatures 0oC
carbon dioxide, increases the directly proportional to the light and 25oC, the rate of
rate of photosynthesis. intensity. photosynthesis approximately
 More CO2 = more CO2fixation =  As light intensity increases, the doubles for each 10oC rise in
more GP = more TP = more rate of photosynthesis increases. temperature.
organic molecules.  Light has 3 main effects:  Above 25oC:
1. Causes stomatato open so o The rate of photosynthesis
CO2 can enter leaves - more levels off and then falls as
carbon fixation. enzymes(ribulose) workless
2. Trapped by chlorophyll efficiently. Proteins
where it excites electrons - (photosystems and
involved in electroncarriers) also
photophosphorylation = denature.
more ATP. o Oxygen more successfully
3. Splits water molecules to competes for rubisco and
produce protons - involved stops it from accepting CO2.
in photophosphorylation = o More water loss from
more ATP. stomata, leading to a
stressresponse in which the
stomata close, limiting the
availability of CO2.
 F214 Module 3: Photosynthesis
(q) describe how to investigate experimentally the factors that affect the rate of photosynthesis
Using a Photosynthometer:
A photosynthometer is used to measure the rate of photosynthesis by collecting and measuring the volume of
oxygen produced in a certain amount of time.
The gas given off by the plant is collected in the
flaredend of the capillary tube forming a gas bubble
and the length of this bubble can be used to calculate
the volume of gas collected.

Volume of gas collected = length of bubble x πr2

The water bath keeps the temperature constant.

Sodium hydrogencarbonate solution is added to the
water in the tube to provide carbon dioxide. The
investigation has to be carried out in a darkened
room, so that the only light available to the plant is
form the light source. If the same apparatus is used
throughout the investigation, the diameter (and
therefore the radius) is constant and we can compare
the rates of photosynthesis by using just the length of
gas bubble evolved per unit time.

1. Fill the apparatus with water by removing the plunger from the syringe. Replace the syringe plunger and gently
push water out of the flared end of the capillary tube, until the plunger is nearly at the end of the syringe and there
are no air bubbles.
2. Place a cut shoot, end upwards, into a test tube containing the same water that the plant has been kept in and
add 2 drops of sodium hydrogen carbonate solution. Stand the test tube in a beaker of water and use a
thermometer to measure the temperature of the beaker.
3. Place a light source as close to the beaker as possible. Measure the distance (d) from the plant to the light source.
Allow the plant to acclimatise.
4. Position the capillary tube over the cut end of the plant and pull the syringe plunger so that the bubble of gas
collected is in the capillary tube near the scale. Measure the length of the bubble and note it down.
5. Gently push the plunger back so that the bubble is expelled and reposition the capillary tube to repeat the
experiment.
 Light Intensity- move the light source further from the plant. Measure the distance and calculate the
light intensity (or use a light meter to measure light intensity). Allow the plant to acclimatise and repeat
steps 4 and 5.
 Temperature- keep all other factors constant and use a light intensity that produced a high rate of
photosynthesis. Alter the temperature of the water bath and measure the volume of gas produced, in a
known period of time, at each temperature.
 Carbon Dioxide Concentration- keep all other factors constant and vary the number of drops of
sodium hydrogencarbonate solution. Measure the volume of gas produced, in a known period of time, at
each temperature.
 F214 Module 3: Photosynthesis
Using Changes in Density of Leaf Discs:
1. Use a drinking straw to cut several leaf discs.
2. Place 5/6 discs in a 10cm3 syringe and half-fill the syringe with dilute sodium hydrogencarbonatesolution.
3. Hold the syringe upright, place your finger over the end of the syringe and gently pull on the plunger - pulls the
air out of the spaces of spongy mesophyll in the leaf disks. As the density of the discs increase they sink to the
bottom of the syringe.
4. Once the discs have sunk, transfer the contents of the syringe to a small beaker. Shine a light from above and
time how long it takes for one leaf disc to float to the top of the solution - the reciprocal of the time taken (1/t) is a
measure of the rate of photosynthesis.
5. Repeat the procedure twice more at this light intensity and find the mean rate of photosynthesis. Repeat at other
light intensities.

This method can also be adapted to investigate the effect of temperature, light wavelength or carbon dioxide
concentration on the rate of photosynthesis.