C1.

3 Photosynthesis
Interaction and interdependence—Molecules
Standard level and higher level: 3 hours
Additional higher level: 3 hours

Guiding questions
• How is energy from sunlight absorbed and used in photosynthesis?
• How do abiotic factors interact with photosynthesis?
Recommended prior learning: C1.2 Cell respiration

SL and HL

C1.3.1—Transformation of light energy to chemical energy when carbon compounds are

produced in photosynthesis

This energy transformation supplies most of the chemical energy needed for life processes in
ecosystems.

Photosynthesis is the conversion of light energy into chemical energy, which is essential to life on earth.
Not only does it produce energy-rich carbohydrates but oxygen as well, both of which have been crucial to
the establishment of complex ecosystems.

C1.3.2—Conversion of carbon dioxide to glucose in photosynthesis using hydrogen obtained by

splitting water

Students should be able to write a simple word equation for photosynthesis, with glucose as the
product.
!"#$%
Water + carbon dioxide /⎯1 glucose + water
Photosynthesis is composed of light-dependent and light-independent reactions.
Light dependent reactions occur in the thylakoids of chloroplasts where light energy is used to split water
in a process called photolysis (“photo” meaning light and “lysis” meaning breaking down). This assists in
converting light energy into chemical energy.
Light independent reactions occur in the stroma of chloroplasts where the chemical energy generated
from the light dependent stages is used to fuel the production of glucose from carbon dioxide.

C1.3.3—Oxygen as a by-product of photosynthesis in plants, algae and cyanobacteria

Students should know the simple word equation for photosynthesis. They should know that the
oxygen produced by photosynthesis comes from the splitting of water.

In plants, algae, and cyanobacteria, oxygen is a by-product of photosynthesis which results from the
photolysis of water.
C1.3.5—Absorption of specific wavelengths of light by photosynthetic pigments

Include excitation of electrons within a pigment molecule, transformation of light energy to

chemical energy and the reason that only some wavelengths are absorbed. Students should be
familiar with absorption spectra. Include both wavelengths and colors of light in the horizontal axis
of absorption spectra.

A photon is a type of electromagnetic radiation that can be seen by humans as visible light. The higher the
energy of a photon, the shorter its wavelength, and vice versa. When electrons within a pigment molecule
like chlorophyll absorb a photon, they become excited/filled with energy and aid in the transformation of
light energy to chemical energy. The amount of energy/wavelength absorbed depends on the chemical
identity of the pigment molecule.

C1.3.4—Separation and identification of photosynthetic pigments by chromatography

Application of skills: Students should be able to calculate Rf values from the results of
chromatographic separation of photosynthetic pigments and identify them by color and by values.
Thin-layer chromatography or paper chromatography can be used.

Pigment A

Pigment B

Origin

Figure 1: paper chromatography.

distance travelled by the pigment (solute)

R & value =
distance travelled by the solvent
C1.3.6—Similarities and differences of absorption and action spectra

Application of skills: Students should be able to determine rates of photosynthesis from data for
oxygen production and carbon dioxide consumption for varying wavelengths. They should also be
able to plot this data to make an action spectrum.

Absorption spectra are graphs depicting the wavelengths absorbed by a chemical substance, like
chlorophyll. The x-axis represents the wavelengths in nanometers and the y-axis shows the amount of light
absorbed from 0-100% or arbitrary units (au).

Figure 2: absorption spectra of the major types of chlorophylls and carotenoids (Johnson).
Action spectra are graphs depicting rate of photosynthesis (y-axis) plotted against wavelengths of visible
light (x-axis).

Figure 3: action spectrum of photosynthesis (Hamblin).

C1.3.7—Techniques for varying concentrations of carbon dioxide, light intensity or temperature
experimentally to investigate the effects of limiting factors on the rate of photosynthesis

Application of skills: Students should be able to suggest hypotheses for the effects of these
limiting factors and to test these through experimentation.

Carbon dioxide: increasing CO2 concentration increases the rate of photosynthesis until another limiting
factor becomes short in supply. CO2 levels can be varied through several methods:

• Using aquatic photoautotrophs and dissolving different amounts of sodium bicarbonate

• In a sealed chamber, CO2 injectors can vary gas concentrations with a sensor to help in monitoring

Light intensity: increasing light intensity increases the rate of photosynthesis until another limiting factor
becomes short in supply.

• Adjusting distance of photoautotroph from light source

• Adjusting light source intensity then measuring light intensity using a lux meter

Temperature: increasing temperature increases rate of photosynthesis up until the optimum temperature is
reached in which any further increase causes denaturation of enzymes like rubisco, decreasing the rate of
photosynthesis.

• Using a water bath and monitoring using thermometer

• Changing temperature of the atmosphere through air conditioning or enclosed chamber

C1.3.8—Carbon dioxide enrichment experiments as a means of predicting future rates of

photosynthesis and plant growth

Include enclosed greenhouse experiments and free-air carbon dioxide enrichment experiments
(FACE).

With the rise in CO2 levels since the industrial revolution, it is important to also understand how this might
not only affect photoautotrophs but the ecosystems in which they exist in as well. Although there is a lot of
research regarding the effects of CO2 on plants, most of these studies have been done in very controlled
labs and greenhouses. While this has allowed us to better understand the physiology of plants, it provides
little insight into how these environmental factors influence ecosystems as a whole.

Free-Air Carbon Dioxide Enrichment (FACE) experiments enable the study of increased CO2
concentrations on plants and ecosystems under natural (free-air) conditions without confinement or
enclosure (not within a greenhouse or lab, where plant competition and other ecological mechanisms are
not considered). This is done through installing CO2 blowers/injectors in certain areas of an ecosystem and
relying on wind to disperse the gas in order to raise the levels of CO2 and observe its effects. Some
hypotheses around these experiments include:

• Increases in CO2 will increase crop yield (result: FACE experiments showed smaller increases in
crop growth than expected)
• Different types of plants will respond more to increased CO2 than others (result: FACE experiments
showed that trees were more responsive than grasses)

Since FACE experiments take years to complete, there is still a lot of data needed before predicting future
rates of photosynthesis and plant/ecosystem growth with sufficient certainty.
NOS: Hypotheses are provisional explanations that require repeated testing. During scientific
research, hypotheses can either be based on theories and then tested in an experiment or be
based on evidence from an experiment already carried out. Students can decide in this case
whether to suggest hypotheses for the effects of limiting factors on photosynthesis before or after
performing their experiments. Students should be able to identify the dependent and independent
variable in an experiment.

An independent variable is the variable being manipulated/changed in an experiment so as to explore its

effects; it is the ‘cause’.
A dependent variable is the variable being measured/observed in an experiment; it is the ‘effect’.
Hypotheses are provisional explanations that require repeated testing. During scientific research,
hypotheses can either be based on theories and then tested in an experiment or be based on evidence
from an experiment already carried out.

NOS: Finding methods for careful control of variables is part of experimental design. This may be
easier in the laboratory but some experiments can only be done in the field. Field experiments
include those performed in natural ecosystems. Students should be able to identify a controlled
variable in an experiment.

Controlled variables remain unchanged/fixed during an experiment to reach valid and precise conclusions.

Aspect Lab experiments Field experiments

Controlling variables Easier and more precise Harder and less precise
High due to standardized equipment Low due to variable natural
Reproducibility
and conditions conditions
Ecological applicability Less ecologically relevant More ecologically relevant
Additional higher level

C1.3.9—Photosystems as arrays of pigment molecules that can generate and emit excited
electrons

Students should know that photosystems are always located in membranes and that they occur in
cyanobacteria and in the chloroplasts of photosynthetic eukaryotes. Photosystems should be
described as molecular arrays of chlorophyll and accessory pigments with a special chlorophyll as
the reaction center from which an excited electron is emitted.

Photosystems are protein complexes embedded within thylakoidal membranes that aid in the conversion
of light energy to chemical energy and are composed of two main components:
• Antenna complex: consists of several light-harvesting protein complexes that contain accessory
pigments (chlorophyll a, chlorophyll b, carotenoids). These pigments protect the special
chlorophylls in the reaction center from oxidation by absorbing sunlight and transfer this light
energy between pigments within the antenna until reaching the reaction center. Thus, the antenna
complex acts as a funnel by harnessing and directing light energy to a site where it can be
effectively used.
• Reaction center: a protein complex that contains a special pair of chlorophyll molecules whose
electrons, once excited by energy passed from accessory pigments, are immediately transferred to
carriers in the Electron Transport Chain (ETC).

There are two types of photosystems (named according to the order in which they were discovered and not
in the order of electron transfer) in thylakoids which have the same structure but differ on the basis of what
they oxidize and reduce.

• Photosystem II (PSII) is the first one involved in electron transfer which oxidizes water (photolysis)
and reduces an electron-carrier molecule (plastoquinone, PQ).
• Photosystem I (PSI) is the second photosystem involved in electron transfer which oxidizes an
electron-carrier molecule (plastocyanin, PC) and reduces NADP+.

Figure 4: structure of a photosystem (Ann Clark).

C1.3.10—Advantages of the structured array of different types of pigment molecules in a

photosystem

Students should appreciate that a single molecule of chlorophyll or any other pigment would not
be able to perform any part of photosynthesis.

A pigment is a molecule whose electrons selectively absorb some wavelengths and reflect others. The
reflected wavelengths that aren’t absorbed mix together to form the pigment’s color. There are several
advantages of the structured array of different types of pigment molecules in a photosystem:

• The diversity of pigments broadens the range of wavelengths that can be absorbed by a
photosystem, so collectively a greater amount of sunlight can be captured
• Some of the pigments help protect against photodamage by dissipating excess energy as heat
• The proximity of pigments to other ones in the antenna complex facilitates efficient and effective
transfer of energy
C1.3.11—Generation of oxygen by the photolysis of water in photosystem II

Emphasize that the protons and electrons generated by photolysis are used in photosynthesis but
oxygen is a waste product. The advent of oxygen generation by photolysis had immense
consequences for living organisms and geological processes on Earth.

When the special chlorophylls in PSII lose their two electrons to an electron carrier (plastoquinone), they
become a strong oxidizing agent. This causes the photolysis of water according to the following equation:

2H' O → O' + 4H( + 4𝑒 )

The protons produced contribute to the proton gradient during chemiosmosis.

The electrons produced compensate for those lost from the special chlorophylls, 2 for each molecule within
the pair.
The oxygen generated is a waste product and leaves the plant through the stomata. The production of
oxygen billions of years ago had immense consequences for living organisms and geological processes on
Earth. Photoautotrophs first evolved inside oceans, so oxygen in water reacted with dissolved iron to
produce iron oxide, which precipitated and formed banded iron formations (rocks). Once iron and other
elements in the ocean were oxidized, oxygen started to accumulate in the atmosphere, leading to the Great
Oxidation Event and the evolution of aerobic respiration.

C1.3.12—ATP production by chemiosmosis in thylakoids

Include the proton gradient, ATP synthase, and proton pumping by the chain of electron carriers.
Students should know that electrons are sourced, either from photosystem I in cyclic
photophosphorylation or from photosystem II in non-cyclic photophosphorylation, and then used
in ATP production.

Once electrons are transferred from the special chlorophylls to the electron-carrier molecules, they move
through the ETC, and their energy is used by cytochrome complexes to pump protons into the thylakoid
lumen. Due to the small size of thylakoids, a steep proton gradient is quickly established between the
stroma (low H+ concentration) and the thylakoid lumen (high H+ concentration). This causes the protons to
passively diffuse into the stroma through ATP synthase via chemiosmosis, which generates the ATP
necessary to fuel the light-independent reactions later on.

The final electron acceptor in the ETC is PSI, which accepts the now low-energy electrons from
plastocyanin into its reaction center. Each of the electrons received are boosted to a high-energy state by
light energy funneled through the antenna complex and then used to reduce NADP+ in the stroma. This
process is called noncyclic photophosphorylation, which produces around 1 ATP molecule per pair of
electrons passed to NADP+.

Cyclic photophosphorylation occurs when the cell needs to produce more ATP molecules to power the
light-independent reactions. Instead of passing the re-energized electrons to NADP+, PSI passes them back
to the cytochrome complexes in order to pump more protons into the thylakoid lumen, thereby increasing
the proton gradient that drives ATP synthase to produce more ATP. The cell has mechanisms to regulate
how much light energy is converted into high-energy phosphate bonds (ATP) and how much into reducing
power (NADPH).

Figure 5: diagram of light-dependent reactions (Ann Clark).

C1.3.13—Reduction of NADP by photosystem I

Students should appreciate that NADP is reduced by accepting two electrons that have come
from photosystem I. It also accepts a hydrogen ion that has come from the stroma. The paired
terms “NADP and reduced NADP” or “NADP+ and NADPH” should be paired consistently.

NADP (nicotinamide adenine dinucleotide phosphate) is the same as a typical NAD molecule but with an
added phosphate group. NADP+ can undergo reduction to gain electrons and a hydrogen ion (NADPH), or
oxidation to donate electrons and a proton. NADP reductase is an enzyme embedded within the thylakoid
membrane that catalyzes the reduction of NADP+ to NADPH. Since a proton is used to reduce NADP+, this
decreases the H+ concentration of the stroma which further increases the steepness of the proton gradient.

C1.3.14—Thylakoids as systems for performing the light-dependent reactions of photosynthesis

Students should appreciate where photolysis of water, synthesis of ATP by chemiosmosis and
reduction of NADP occur in a thylakoid.

Thylakoids are membrane-bound compartments within chloroplasts that perform the light-dependent
reactions of photosynthesis. A stack of disc-shaped thylakoids is called a granum (plural grana), which is
connected to other grana via lamellae (unstacked stroma thylakoid).

PSII are mainly concentrated in grana and PSI in lamellae. Since lamellae have more access to the stroma
than grana, NADP+ reduction is easier in them compared to grana.

C1.3.15—Carbon fixation by Rubisco

Students should know the names of the substrates RuBP and CO2 and the product glycerate 3-
phosphate. They should also know that Rubisco is the most abundant enzyme on Earth and that
high concentrations of it are needed in the stroma of chloroplasts because it works relatively
slowly and is not effective in low carbon dioxide concentrations.

The Calvin Cycle constitutes the light-independent reactions of photosynthesis and involves 3 main steps:

1. Carbon fixation
2. Reduction
3. Regeneration

CO2 is a non-polar and small molecule that can easily diffuse through stomatal openings into plant cells.
This makes it a suitable provider of the carbon atoms needed as backbone for biomolecules like glucose.

In carbon fixation, the enzyme Rubisco catalyzes the reaction between CO2 molecule and ribulose
bisphosphate (RuBP), a 5C compound with 2 phosphate groups, which produces 2 glycerate-3-
phosphate (GP) molecules.

Rubisco is the most abundant enzyme on earth and found in all three domains of life. It works relatively
slowly compared to most enzymes, which according to evidence may be because of the reciprocal
relationship between enzyme specificity and activity. Ever since the great oxidation event, the enzyme had
to evolve to precisely distinguish between CO2 and O2 molecules, which came at the cost of a reduced
catalytic rate. The enzyme is also not very effective at low CO2 concentrations, so high concentrations of
Rubisco are needed within the stroma of chloroplasts.

C1.3.16—Synthesis of triose phosphate using reduced NADP and ATP

Students should know that glycerate-3-phosphate (GP) is converted into triose phosphate (TP)
using NADPH and ATP.

In the reduction step, the 2 GP molecules are reduced into triose phosphate (TP), a 3C compound, by
hydrolyzing 2 ATP molecules into ADP + Pi and oxidizing 2 NADPH molecules into NADP+.
C1.3.17—Regeneration of RuBP in the Calvin cycle using ATP

Students are not required to know details of the individual reactions, but students should
understand that five molecules of triose phosphate are converted to three molecules of RuBP,
allowing the Calvin cycle to continue. If glucose is the product of photosynthesis, five-sixths of all
the triose phosphate produced must be converted back to RuBP.

In the regeneration step, and for every 3 Calvin cycles, 1 TP molecule is used to make glucose while the
other 5 are used to regenerate RuBP to enable sustainable production. 3 ATP molecules are hydrolyzed
into 3ADP + 3Pi in this step per 3 molecules of CO2 fixed.

Since 6 carbon atoms make up the structure of glucose, 6 CO2 molecules need to be fixed, so 6 Calvin
Cycles need to be performed in order to produce 1 glucose molecule.

Figure 6: 3 turns of the Calvin cycle (Molnar).

C1.3.18—Synthesis of carbohydrates, amino acids and other carbon compounds using the
products of the Calvin cycle and mineral nutrients

Students are not required to know details of metabolic pathways, but students should understand
that all of the carbon in compounds in photosynthesizing organisms is fixed in the Calvin cycle
and that carbon compounds other than glucose are made by metabolic pathways that can be
traced back to an intermediate in the cycle.

All of the carbon in compounds in photosynthesizing organisms is fixed in the Calvin cycle. Other carbon
compounds, like amino acids and lipids, are made by metabolic pathways that can be traced back to an
intermediate in the cycle. For example, amino acids can be synthesized using TP and fatty acids using GP
with the addition of phosphates and sulfurs from mineral nutrients like ammonium.
 C1.3.19—Interdependence of the light-dependent and light-independent reactions

Students should understand how a lack of light stops light-dependent reactions and how a lack of
CO2 prevents photosystem II from functioning.

The light-dependent and independent reactions are interconnected and interdependent on each other. If the
light-dependent reactions do not occur, the Calvin cycle cannot proceed as it has no ATP and NADPH to
fuel it, and if the Calvin cycle does not take place, then the light-dependent reactions will not have the
biomolecules it needs to function.

Lack of CO2 prevents PSII from functioning, because there is no need to produce more ATP and NADPH if
there is no carbon dioxide for the Calvin cycle to fix into carbohydrate.

Linking questions
• What are the consequences of photosynthesis for ecosystems?
• What are the functions of pigments in living organisms?

Review questions
SL and HL
• Identify the reactants and products of photosynthesis. [1]
• Outline the consequences of constantly increasing light intensity past the saturation point. [2]
• Explain the significance of photosynthesis to life on earth. [3]
• Outline why photosynthesis efficiency might differ between a plant growing at sea level and one
at high altitudes. [3]
• Explain the importance of FACE experiments in predicting the effects of climate change on both
plants and ecosystems. [4]

Additional higher level

• State why photosystem II does not function when carbon dioxide is missing. [1]
• Suggest why two forms of energy, ATP and NADPH, are needed during the Calvin cycle. [1]
• Outline why carbon dioxide is a good source of carbon in photosynthesis. [2]
• Outline the interdependence of the light-dependent and independent reactions of
photosynthesis. [2]
• Explain the role of rubisco in photosynthesis. [3]
• Outline the role of accessory pigments in photosynthesis. [3]
• Explain how the proton gradient across the thylakoid membrane influences the synthesis of ATP
during photosynthesis. [3]
• Explain the consequences of releasing oxygen as a by-product of photosynthesis. [3]
• Explain the role of reduced NADP in photosynthesis. [4]
• Describe how a proton gradient is established between the thylakoid lumen and chloroplast
stroma. [4]
• Explain the significance of photolysis in photosynthesis. [4]
• Explain the role of ATP in photosynthesis. [4]
• Compare and contrast the two types of photophosphorylation in the light-dependent reactions of
photosynthesis. [4]
• Explain the significance of having two variations of the electron transport chain in thylakoids. [4]
• Describe the Calvin cycle. [4]
• Explain how the structure of a chloroplast is adapted to its function. [5]
• Compare and contrast the photosystems involved in photosynthesis. [6]
• Explain how photosystems I and II are adapted to their function. [6]
• Explain how variations in chlorophyll concentrations can affect the absorption spectrum of a leaf,
and outline how this might influence a plant's ability to adapt to different light environments. [7]
• Discuss how ATP is produced within chloroplasts. [8]
• Discuss the movement of electrons during the light-dependent and independent reactions of
photosynthesis. [8]
References
Ainsworth, Elizabeth A., et al. "What have we learned from 15 years of free-air co2 enrichment (face)? a meta-analytic
review of the responses of photosynthesis, canopy properties and plant production to rising co2". New Phytologist, vol.
165, no. 2, 2004, p. 351-372. https://doi.org/10.1111/j.1469-8137.2004.01224.x.

Ann Clark, Mary, et al. Biology 2e. E-book, OpenStax, 2018, https://openstax.org/books/biology-2e/pages/1-introduction.
OpenStax.

Erb, Tobias J, and Jan Zarzycki. “A short history of RubisCO: the rise and fall (?) of Nature's predominant CO2 fixing
enzyme.” Current opinion in biotechnology vol. 49 (2018): 100-107. doi:10.1016/j.copbio.2017.07.017.

Gordon Betts, J., et al. Anatomy and Physiology 2e. E-book, OpenStax, 2022, https://openstax.org/books/anatomy-and-
physiology-2e/pages/1-introduction. OpenStax.

Hamblin, Michael & Huang, Ying-Ying & Heiskanen, Vladimir. (2018). Non-mammalian Hosts and Photobiomodulation: Do
All Life-forms Respond to Light?. Photochemistry and Photobiology. 95. 10.1111/php.12951.

Johnson, Matthew P. (2016). Photosynthesis. Essays In Biochemistry. 60. 255-273. 10.1042/EBC20160016.

Molnar, C., & Gair, J. (2015). Concepts of Biology – 1st Canadian Edition. BCcampus. Retrieved from
https://opentextbc.ca/biology/.

Rojdestvenski, Igor et al. “Segregation of photosystems in thylakoid membranes as a critical phenomenon.” Biophysical
journal vol. 82,4 (2002): 1719-30. doi:10.1016/S0006-3495(02)75524-0.