BIO-RAD TM

Photosynthesis and Cellular Respiration Kit

A ThINQ!™ Investigation
Catalog #17001238EDU

AP Biology Student Manual

Dear Students
Take a deep breath in and out. What is your body
doing when you inhale and exhale? Did you say
absorbing oxygen (O2) and releasing carbon dioxide
(CO2)? But where does this O2 come from and
where does the CO2 go? Why doesn’t the earth’s
atmosphere run out of O2 and accumulate CO2?
Hopefully you’ll be able to answer these and
many other questions after investigating
photosynthesis and cellular respiration through
a series of experiments.

This instruction manual, and the experiments

outlined within it, will help you understand
the importance of photosynthesis and
cellular respiration to all life on Earth: from
the algae in your fish tank to the birds in
the trees to the great white sharks in the
ocean. While exploring these subjects you
will have the opportunity to ask questions
and search for answers to these questions.
Sometimes you will find the answers and at other
times you will discover more questions; both will
contribute to your knowledge.

You may apply this knowledge by considering how our

actions as human beings affect the balance of photosynthesis
and cellular respiration and atmospheric O2 and CO2. How are
greenhouse gases contributing to changes in the ocean’s conditions
and the species that can live there? How might replacing rainforests with
farms affect the local ecosystem? How are local farming practices affecting other
geographic regions? What are the benefits and detriments of our daily decisions?
We hope that you will form your own opinions and perhaps suggest improvements
that we as a human race can make. Most importantly, we hope you stay curious
about the world around you and never stop asking questions.

Bio-Rad’s Explorer Team

Bio-Rad Laboratories
6000 James Watson Drive, Hercules, CA 94547
[email protected]
Introduction
Photosynthesis and Cellular Respiration
In the following lab investigations, you will use algae beads (algae cells that are encapsulated ThINQ!
in alginate) and a colorimetric CO2 indicator to observe two biochemical processes that
sustain most life on Earth: photosynthesis and cellular respiration. It’s not an exaggeration
Exercises
Collaborate and use outside
to say that we owe our lives to these two processes. Photosynthesis converts the energy in
resources to answer the
sunlight into the sugars we eat and the oxygen we breathe; it is through cellular respiration
following questions:
that we are able to use the sugars and oxygen to sustain our lives.
When did molecular
Through a series of inquiry experiments and pre- and post-lab activities, you will explore how oxygen (O2 ) first appear in
these two processes share mechanistic and evolutionary ties, and how all life on Earth is tied Earth’s atmosphere? What
inextricably through them. This will lay a foundation for understanding how imbalances in Earth’s organism and processes led
resources can upset the balance of these processes, leading to far-reaching consequences. to O2 accumulation in our
This introduction provides a general overview to photosynthesis and cellular respiration. atmosphere?

Organisms need matter and energy (food) to live

All living things need food to grow and to survive. The food we eat supplies us with the
matter (“stuff”) we need to make more of ourselves, and it supplies the energy we need to
fuel our lives. Our lives, then, follow the first law of thermodynamics, which states the
universe holds a finite amount of energy and, as a result, energy can be neither created nor
destroyed. It can, however, be captured, transferred, and converted into other forms. We
cannot simply “make” the matter and energy we need; we must get it from somewhere. In Where does most of Earth’s O2
come from today (for example,
fact, all life on Earth is based on systems of energy capture, transfer, and transformation.
oceans, rainforests, etc.)?
Life also follows the second law of thermodynamics, which states that during any energy
transformation, a portion of the energy is lost to entropy. Entropy is the measure of disorder
within a system. All things tend to move to a state of higher entropy, and energy input is
required to maintain order and organization. To survive, living things must offset entropy by
taking in more energy than they lose or expend. Otherwise, harmful or even fatal energy
deficiencies can occur. This can affect population size and even cause disruptions at the
ecosystem level.
If photosynthesis captures
Organisms need energy to survive only 1–2% of the energy from

Photosynthesis and cellular respiration are great examples of energy-transforming systems, sunlight, what happens to the
other 98–99% of energy?
and they define the two ways in which organisms derive the energy they need to survive.
Autotrophs (also called primary producers) capture free energy from the environment,
including energy from sunlight (photosynthesis) or chemical sources (chemosynthesis). They
transform this energy into other forms that can be used by themselves and other organisms
within their environment. Some examples of autotrophs include plants, algae, and some Photosynthesis and
chemosynthesis are two forms of
bacteria. Heterotrophs (consumers), on the other hand, obtain free energy from carbon
autotrophy. Provide an example
compounds produced by other organisms, including other heterotrophs and autotrophs.
of a chemosynthetic organism.
Within the web of life, autotrophs provide the food needed by all heterotrophs to grow,
survive, and reproduce.

Energy is captured in universal units of chemical currency

The biochemical pathways that organisms use to capture energy occur in a stepwise
fashion. At each stage energy is trapped in the form of energy-storing chemicals (“chemical
currency”). For example, organisms can burn sugar through cellular respiration and store
the energy as the following four chemicals until it is needed to perform other types of work: Lichens represent a symbiotic
relationship between a
ATP (adenosine triphosphate) — this molecule contains a high-energy phosphate bond.
fungus and an alga. In this
When the molecule is hydrolized, this bond is broken, and the energy released can be
relationship, which organism
coupled to other reactions that require energy to proceed.
is the autotroph? Which is the
ATP —> ADP + Pi (energy utilization reaction) heterotroph?

NADPH (nicotinamide adenine dinucleotide phosphate), NADH (nicotinamide

adenine dinucleotide), FADH2 (flavin adenine dinucleotide) — these three compounds
store energy that is used in oxidation/reduction reactions.

1
Photosynthesis and cellular respiration are also the source of the stuff we use, such as
shampoo, plastic water bottles, clothes, and even medications. The materials for these items
ThINQ!
are derived from petroleum, which is a mixture of hydrocarbons formed by the compression
of ancient fossilized organisms on the ocean bed. These ancient organisms were autotrophs Exercises
and heterotrophs that depended on photosynthesis to convert atmospheric CO2 to energy Collaborate and use outside
and organic matter. The process of converting CO2 to organic matter, then organic matter resources to answer the
to petroleum (hydrocarbons) can be viewed as energy storage and CO2 capture from the following questions:
atmosphere. What is happening to the captured CO2 when we convert petroleum to energy In photosynthesis, water (H2O)
and the things we use in our daily lives? is the ultimate electron donor,
and it is split to yield O2 . Provide
Photosynthesis and Cellular Respiration Are Interdependent Pathways an example of an electron donor
That Are Central to Life used in chemosynthesis. What
In one way or another, all life on Earth depends on photosynthesis and cellular respiration. is the byproduct of its electron
Photosynthesis is the only biological process that can capture energy from sunlight and donation?

convert it into chemical compounds that all organisms — from bacteria to humans — use to
power metabolism, growth, and reproduction. Cellular respiration, in turn, is the process all
organisms require to derive energy from the products of photosynthesis (for example, sugars)
they consume. The carbohydrates produced by photosynthesis can be used to drive multiple
different metabolic processes, including cellular respiration. Cellular respiration uses the free
energy from sugar, for example, to produce a variety of metabolites and to phosphorylate ADP
into ATP to fuel other processes.

Although photosynthesis and cellular respiration evolved as independent processes in early

prokaryotes, a look at the summary reactions (see figure below) highlights their interdependence
today: The products of photosynthesis — oxygen and carbohydrates — are the reactants for
cellular respiration, and vice versa.

Photosynthesis: What do synthase enzymes do?

6 CO2 + 6 H2O + light (energy) → C6H12O6 (sugar) + 6 O2 What are the two precursors that
ATP synthase uses as reactants
to produce ATP?
O2

Chloroplast
CO2 + H2O Sugars
Mitochondrion

O2
Energy
Cellular Respiration:
C6H12O6 (sugar) + 6 O2 → 6 CO2 + 6 H2O + 36 ATP (energy)

Photosynthesis Occurs in Two Phases Within chloroplasts

Photosynthesis powers 99% of Earth’s ecosystems; it is through the photosynthetic activities of plants, algae, and certain bacteria
that our atmosphere gains the molecular oxygen we breathe and the sugars we eat. In addition, photosynthesis helps maintain the
balance between oxygen and carbon dioxide in our atmosphere. This balance is so important that scientists are trying to create
artificial photosynthetic systems to capture CO2 emissions from factories and coal-burning energy plants before they are released
into the atmosphere. Photosynthesis is even responsible for fueling our cars — the energy stored within fossil fuels (natural gas,

explorer.bio-rad.com 2
coal, petroleum, etc.) is derived from solar energy that was trapped and stored during
photosynthesis long ago in the geological past. Today the global rate of energy capture
ThINQ!
through photosynthesis is approximately three times the rate of power consumption by
humans. Photosynthesis occurs as two separate but coordinated sets of reactions: Exercises
Collaborate and use outside
• The Hill reaction, in which the energy from sunlight is captured and converted to
resources to answer the
chemical energy and water (H2O) is split to form free oxygen (O2)
following question:
• The Calvin cycle, in which the ATP and NADH made in the Hill reaction are used
The carbon fixation reactions
to convert CO2 to sugar
are sometimes called the dark
reactions to distinguish them
In eukaryotes, these reactions take place in an organelle specialized to the task: the from the Hill reaction. Is this an
chloroplast. The structure of the chloroplast facilitates its function in photosynthesis. accurate name? In other words,
Stacked and folded inner membranes (thylakoids) contain the pigments (chlorophylls and do these reactions occur only in
carotenoids) and enzymes required for the Hill reaction; the folded and stacked structures the dark?
provide a large surface area for pigments and light harvesting. In prokaryotes, these reactions
occur in infoldings of the inner membrane. The carbon fixation reactions occur within the
stroma of the chloroplast (cytosol of prokaryotes).

Directly or indirectly, nearly all ecosystems on Earth are powered by photosynthesis. For
example, when a top predator, such as a coyote, preys on a rabbit, the coyote is at the end
of an energy path that originated with nuclear reactions on the surface of the sun to light to
photosynthesis to vegetation to the rabbits and, finally, to the coyote.

Hill Reaction Involves an Electron Transport Chain

During the Hill reaction, chlorophylls and other pigments capture the free energy from
sunlight and convert it to a higher, more excited energy state. Excited chlorophylls pass
electrons down an electron transport chain to successively lower energy states, creating
reduced intermediates along the way. This electron transport chain ultimately yields:

• NADPH (one of the forms of universal energy currency mentioned above)

• An electrochemical gradient of protons (hydrogen ions, H+) across the thylakoid
membrane; this gradient drives the synthesis of ATP by the enzyme ATP synthase,
which converts the energy stored in the electrochemical gradient into the energy of ATP
• A chlorophyll molecule without an electron. Chlorophyll regains this electron through the
splitting of water (H2O), which also yields O2

Hill net reaction: 2 H2O + 3 ADP + 3 Pi + 2 NADP+ + energy (light) —> 3 ATP + 2 NADPH + O2

The NADPH and ATP are energy molecules that are utilized in the Calvin cycle to make sugars; the oxygen that is generated fuels
respiration by aerobic organisms like us.

Light Chloroplast
The Hill CO2
reaction in Stroma
the thylakoid
membrane Thylakoid P+
NADP
N
membrane AD + Pi
ADP
A
produces O2, The Calvin
ATP, and cycle in the
NADPH. H2O Calvin
cycle stroma uses
Hill
O2 reaction
CO2, ATP,
ATP
A P and NADPH
NADPH
N PHH
to make
carbohydrates,
O2 such as sugars.
Sugars
Cytosol

3
The Calvin Cycle Is Powered by the Products of the Hill Reaction
The Calvin cycle (carbon-fixing) reactions of photosynthesis use the ATP and NADPH made during the Hill reaction to convert CO2
into sugars.

Calvin net reaction: 6 CO2 + 18 ATP + 12 NADPH —> C6H12O6 (glucose) + 12 NADP+ + 18 ADP + 18 Pi

The enzyme RuBisCO (Ribulose Bisphosphate Carboxylase Oxygenase) catalyzes the CO2 fixation step of the Calvin cycle.
Highlighting its critical role in sustaining life, RuBisCO is the most abundant protein on Earth — ~20–50% of the protein in every
leaf is RuBisCO.

The sugar and other intermediates produced by photosynthesis are the branch points for a number of different metabolic
pathways, including those that synthesize fats, amino acids, and other critical building blocks of life. Glucose also serves as a vital
food source and as a primary entry point to glycolysis and cellular respiration.

Reactions of Glycolysis and Cellular Respiration Release Energy from Food

Where photosynthesis uses sunlight, water, CO2, ADP, and NADP+ as input and generates the sugars we eat and O2 we breathe,
glycolysis and cellular respiration essentially reverse these processes: aerobic organisms eat compounds like glucose and, in the
presence of oxygen that is breathed, convert the glucose to CO2 and water and extract a huge amount of energy in the process.

Mitochondrion Chloroplast Bacterium (E. coli)

Matrix Stroma Cytosol

ATP ATP
H+ H+

H+
ATP
Intermembrane Thylakoid Intermembrane space
space lumen (periplasm)

And as with photosynthesis, the reactions of glycolysis and cellular respiration are compartmentalized in eukaryotes: In eukaryotes,
glycolysis (the initial breakdown of sugar) occurs in the cytosol, and cellular respiration — which also involves an electron transport
chain and an ATP synthase — occurs within mitochondria.

In fact, as we sleep each night, we lose weight just from respiration. It is estimated that, for every gram of air we breathe in as we sleep,
we lose about 0.013 gram of carbon (in CO2) and 0.019 gram of water vapor. This can add up to a pound of weight loss overnight!

Aerobic Respiration of Glucose

C6H12O6 (glucose) + 6 O2 —> 6 CO2 + H2O + energy

In the presence of oxygen, the stepwise combustion of glucose yields energy in the forms of ATP, NADH, and FADH2. Ultimately,
since the latter two can be converted to units of ATP, one molecule of glucose yields approximately 36 molecules of ATP!
Remember, ATP is used for many different processes, including muscle action, nerve impulses, and other metabolic processes . . .
a lot of them!

explorer.bio-rad.com 4
Photosynthesis and Cellular Respiration Occur within the Same Cell
It is important to understand that, although only autotrophs perform photosynthesis, ThINQ!
ALL organisms (you, your teacher, the neighbor’s cat, and the tree at the end of the street)
perform glycolysis and cellular respiration. In fact, the reactions that break down glucose
Exercises
in the presence of oxygen are universal. Even autotrophs, who produce their own food, Collaborate and use outside
use glycolysis and cellular respiration to break down the sugars they synthesize in order to resources to answer the
extract energy and metabolites along the way. Where photosynthesis is the capture and following question:

transformation of light energy to chemical energy (photosynthates), respiration is the burning In the absence of oxygen,
of those photosynthates for energy to grow and to do the work of living. Both plants and organisms may undergo
animals (including microorganisms) need oxygen for aerobic respiration. This is why overly wet anaerobic fermentation to
or saturated soils are detrimental to root growth and function, as well as to the decomposition extract chemical energy and
processes carried out by microorganisms in the soil. metabolic intermediates from
glucose. What are some of the
products of fermentation that
In autotrophs such as algae, these pathways occur within the same cells! In fact, if you
occur in nature?
could look inside one of the algal cells you will be using in the lab investigations, you’d
see a large central chloroplast as well as smaller mitochondria — all within the same cell.
Though photosynthesis and cellular respiration are connected through common intermediate
metabolites in the cytosol, elegant regulatory pathways and differences in resource availability
ensure the algal cells balance the rates of photosynthesis and cellular respiration as needed to
survive environmental changes.

The algae beads used in the following investigations allow you to observe both pathways
simultaneously. You will incubate the algae beads in a CO2 indicator solution that is sensitive to
changes in pH caused by gaseous CO2 dissolving in water to form carbonic acid:

CO2 + H2O <—> H2CO3 <—> HCO3− + H+

When the CO2 levels are high, the CO2 indicator will turn yellow, and when CO2 levels
decrease, it turns purple.

Summary
The following table summarizes some of the hallmarks of photosynthesis and cellular respiration.

Table 1. Hallmarks of photosynthesis and respiration.

Photosynthesis Cellular Respiration

Input CO2, H2O, light energy Sugars, O2

Output Sugars, O2 CO2, H2O, chemical energy

Organism type Autotrophs (producers) only Autotrophs and heterotrophs

Organelle Chloroplast Mitochondrion

Electron transport In thylakoid membranes In cristae (inner membrane of mitochondria)

chain?

Terminal electron NADP+ to generate NADPH2 for CO2 fixation Oxygen (O2) to generate water (H2O)
acceptor

Requires light? Yes No

5
 Investigations #1 and 2: Scenedesmus obliquus and Examining
the Rates of Photosynthesis and Cellular Respiration
Scenedesmus obliquus is a eukaryotic microalgae. In these investigations, you will observe its
physiology and monitor the overall rates of photosynthesis and cellular respiration under light and
dark conditions.

Pre-lab questions:
1. In your own words (or using chemical reactions), describe how photosynthesis and
cellular respiration are interdependent:
2. What type of organism would you need to use to be able to monitor both
photosynthesis and cellular respiration?
3. Why are the eukaryotic algal cells in the Photosynthesis and Cellular Respiration
lab a good choice?
4. Which process (photosynthesis, cellular respiration, or both) do the algae perform
when incubated in the light? In the dark?
5. Photosynthesis uses CO2 and cellular respiration produces CO2. We call the point
when the two processes are in balance— when there is no net production of
CO2— the compensation point. How might you limit one of the processes in order
to achieve a compensation point?
6. Examining the data below, how do you expect the rate of cellular respiration to
impact the rate of photosynthesis that you can measure in the light and the dark?

7. What would happen to life on Earth if the rates of photosynthesis and cellular
respiration in all phototrophs were equal?
 Investigation #1: Algae Microscopy
Overview
In this exercise, you will depolymerize the algae bead to free the algae (Scenedesmus obliquus)
and observe the algae under a microscope.
The Scenedesmus genus is one of the most common unicellular freshwater algae. Though it
can exist in a single cell (unicell) stage, it is also often seen in coenobia of four to eight cells.
The coenobia you observe may have end cells with two long spines protruding from the outer
corners. Each cell contains a single, plate-like chloroplast. Scenedesmus is used as an
experimental system in research on pollution, photosynthesis, and biofuels. In another practical
application, Scenedesmus provides oxygen for the bacterial decomposition of organic matter in
sewage purification processes.
Protocol
1. Gently invert the tube of debeaded algae to mix, and then use a fresh transfer pipet to
transfer 1 drop of dissolved algae bead solution to the center of a microscope slide.
2. Place a coverslip over the microscope slide.
3. Observe under a microscope, taking notes and making sketches.
Data Collection and Analysis
1. Draw some of the cells you see. Do you see coenobia? If so, how many cells do you
typically see per coenobium?
2. Do the S. obliquus vary in color? What does the intensity of the color tell you about the
algae?
3. Draw a diagram that represents the interdependence of photosynthesis and cellular
respiration within the algae cells. This diagram should include the connections between
the products and reactants for each process, the location/organelle in which each
process occurs, and a short description of what is occurring during photosynthesis and
cellular respiration.

Investigation #2: Photosynthesis and Cellular Respiration

Core Lab Overview
All life on Earth ultimately relies on two biochemical processes: photosynthesis and cellular
respiration. In this exercise, you will use algae beads to measure rates of photosynthesis and
cellular respiration. The beads contain eukaryotic microalgae (Scenedesmus obliquus)
encapsulated in alginate. You will incubate them in a CO2
indicator solution that is sensitive to changes in pH caused by gaseous CO2 dissolving in water
to form carbonic acid:CO2+ H2O <—> H2CO3<—> HCO3−+ H+When the CO2 indicator is at
equilibrium with the atmosphere, it is dark orange. When the CO2 levels increase, it changes to
yellow, and when CO2 levels decrease, it changes to purple (see Indicator Color Guide). The
CO2 indicator spans the range of pH change that will be seen in the algae beads (pH 6.9–9.1),
making it a convenient way to measure photosynthesis and cellular respiration. In this exercise,
you will compare the rates of color change of the CO2 indicator caused by algae beads
incubated under bright light and in complete darkness. The color/pH change of the CO2 indicator
can be determined using the Indicator Color Guide or a spectrophotometer set to measure
absorbance at 550 nm. Indicator Color Guide and corresponding pH values.
Prelab questions
1. As the algae photosynthesize, how will the pH of the CO2 indicator change? Why?
2. How will the pH change if the cells begin to respire?
3. Imagine that the algae are experiencing the light conditions that would result in the graph
from prelab question 6, Predict what color changes will happen in the CO2 indicator
between compensation points 1 and 2, and explain why. What about after compensation
point 2?

Protocol
1. Label one empty cuvette light, and the other cuvette dark. Label each cuvette so that it does
not obstruct light reaching the algae beads.
2. Label a transfer pipet algae and convert it into a scoop (see example) Use the algae transfer
pipet to transfer _____ algae beads into each of the light and dark tubes.
3. Label a new transfer pipet excess and use it to remove and discard the liquid that transferred
along with the beads.
4. Label a new transfer pipet wash and use it to add ____ml of distilled water to each of the
tubes. Let the algae beads incubate in the water for 5 min to allow indicator within the bead to
wash out.
5. Use the wash transfer pipet to remove the water from the tubes. Discard the water into the
waste container.
6. Label a new transfer pipet indicator and use it to transfer ____ ml of CO2 indicator to each
tube. Cap tubes tightly.
7. Wrap the tube labeled dark in aluminum foil. Place both the cuvettes labeled light and dark
on their sides 15–25 cm from the lamp. Ensure that the beads are distributed evenly throughout
the tube and the clear side of the tube faces the light.
8. Collect data starting at time = 0 min. Every 5 min, thoroughly mix the CO2 indicator in the
cuvettes and determine the color. This can be done by comparing the color of the CO2 indicator
in your cuvette to the provided Indicator Color Guide, or by reading the absorbance at 550 nm
(A550) in a spectrophotometer (make sure your teacher has zeroed the machine). Be quick
about taking this reading and immediately return the cuvettes to the experimental conditions.
9. If enough time remains after the last time point, switch the light and dark cuvettes. Place the
cuvette labeled light in the dark and the cuvette labeled dark in the light. Continue to record pH
or A550 every 5 min
Data Collection
1. Enter your data in a table
2. Make some general observations about your experimental setup (type of light bulb, light bulb
color, brightness of the light bulb, distance of your tubes from the light, temperature of the room,
location of your experimental setup relative to other light sources, etc.). It might be useful to
sketch your experimental setup. Why might these general observations be important?

Analysis of Results
The goal of this analysis is to determine the rates of photosynthesis and cellular respiration in
the light and in the dark.
1. Graph your results. Label the y-axis with pH or A 550 value intervals that are appropriate
to your data. Plot the color change versus time for both your light and dark samples on
the same graph, as below. Use a ruler to draw a best fit line for the linear region of your
light and dark datasets. Hint: use a different color to plot your light and dark results, or
use solid and dashed lines.
2. Calculate the slope. Mark two points along your light best fit line. Try to choose points
that are far apart but are still in the linear range of the graph. Do the same with the dark
best fit line. Are your slopes positive or negative for light and dark conditions? What does
this mean about the change in CO2
3. Under which condition did the CO2 indicator turn more alkaline? Why?
4. Under which condition did the solution start to change color more quickly (light or dark)?
(Hint: look at the absolute value of the slopes you calculated.)
5. How does cellular respiration impact the observed rate of photosynthesis? Is your
calculated rate of photosynthesis accurate? Why or why not?
6. Revisit your diagram of photosynthesis and cellular respiration from either Pre-Lab #3 or
Investigation #1. Revise your diagram to demonstrate how the experimental design of
your current investigation affects these processes. Indicate where the rates of these
processes may increase or decrease and describe why. Also indicate how the organism
will be affected if the conditions of your investigation were to continue.
7. Look up current ocean pH values. How do the current values compare to those from
previous years? Consider what you’ve just learned about algae and how the chemistry of
the indicator used in the experiments you just performed works. Hypothesize why oceans
are at their current pH. How is the pH of the ocean changing and why? How might this
affect the organisms that live in the ocean?

Investigations #3–6: Examining Rates of Photosynthesis

and Cellular Respiration under Various Conditions
Overview
Plants cannot move from place to place like animals can when their environment changes
or becomes inhospitable. When temperatures or light availability change, or even when
predators attack, plants have no choice but to stay where they are and survive as best they
can. Plants have adapted to a wide range of habitats, from the very cold and shaded to the
very bright, hot, and humid. Plants have adopted a variety of strategies to accommodate
changing environmental conditions, but how do environmental fluctuations affect rates of
photosynthesis and cellular respiration?
As biochemical processes, photosynthesis and cellular respiration are affected by an array
of biotic and abiotic factors, including the availability of substrates, availability of light, and
the surrounding temperature. In the activities that follow, you will use the algae beads, CO2
indicator, and other supplies to design and perform your own experiments to examine some
of the factors that may influence rates of photosynthesis and cellular respiration.

Effects of Light Quantity and Quality

Sunlight is the starting point for photosynthesis, and the quantity and quality of light available
to a plant — or alga — can fluctuate widely over both time and space. How do you think the
quantity and quality of light might affect plant growth?
Light Intensity
When we talk about light quantity, we are talking about light intensity, the amount of light
hitting a plant leaf. Light intensity also refers to the degree of brightness the leaf might be
exposed to, with high light intensity being brighter than low light intensity.
Light intensity varies significantly in nature. When you plant a garden, you select plants that
can thrive in the various conditions in your yard. Plants are often classified as sun plants
or shade plants based on the light intensity to which they are best adapted. In addition,
individual leaves of a tree often show developmental adaptations to different conditions:
Leaves growing on the exterior of the tree canopy, for example, may exhibit differences in
anatomy, shape, and metabolism (such as photosynthesis) from leaves growing within the
crown of the tree, where they are shaded by surrounding leaves. Light intensity also changes
with the time of day, the season, geographic location, distance from the equator, and weather.
For example, light intensity increases from sunrise to the middle of the day and then decreases;
it is high during summer, moderate in spring and fall, and low in winter. Geographically,
maximum light intensity occurs at the equator and decreases with increasing proximity to the
poles. Light intensity is also affected by dust and water particles in the air.
Light quality (color)
Where light intensity refers to the amount of light available, light quality refers to the
wavelength or color of light available. As a source of energy, visible light occupies only a small
portion of the electromagnetic spectrum (see figure below), which extends from high-energy
gamma rays at one end to lower-energy radio waves at the other. Life on Earth is made possible
by our atmosphere, which filters out most of the higher-energy, ionizing radiation (<295 nm) that
would be hazardous to living organisms.
Light energy must be absorbed by a pigment in order to have a biological effect. In the
plants, algae, and cyanobacteria that are capable of oxygen-producing photosynthesis, the
primary pigment molecule used for capturing sunlight is chlorophyll a. Six other forms of
chlorophylls also exist in nature, along with other accessory pigments that help increase the
range of light that can be harvested. Each of these pigments has a characteristic absorption
spectra (see figure below). For example, chlorophyll and carotenoid pigments are optimized
for absorption of blue and red wavelengths of light

As with light intensity, the growth of plants is strongly correlated to the wavelengths of light
available to them. A number of developmental changes result from changes in light quality,
including flowering and senescence. In the laboratory or classroom, wavelengths of light
from tungsten or fluorescent bulbs are different from those of natural sunlight. As a result,
the rates of photosynthesis measured under artificial light sources will be different from those
observed under the sun.
Effects of Other Environmental Factors
Plants in your garden are sensitive to other environmental factors as well, including
water availability, soil conditions, nutrient availability, presence of pests and disease, and
temperatures. The ability of algae to perform photosynthesis and cellular respiration is
similarly sensitive to environmental factors, including substrate availability (CO2, O2, water)
and temperature. In the laboratory investigations that follow, you will design and conduct
experiments to test the effects of different conditions on photosynthesis and cellular respiration.
The following sections will help you ask a question about photosynthesis and cellular
respiration, formulate a hypothesis, develop an experiment to test this hypothesis, and predict
results.

Investigation #3: Effect of Light Intensity

In nature, light intensity can vary significantly, depending on the time of day or year, location,
climate, and a host of other factors. In the laboratory, neutral density filters, which reduce
transmittance of all wavelengths of light, can be used to manipulate light intensity in a
controlled manner. In this investigation, you will design and conduct an experiment using
neutral density filters to understand the impact of light intensity on the rate of photosynthesis
by the algae beads. For this investigation, all the materials and equipment from Investigation #2
will be made available to you as well as the following:
• Neutral density filters that block 0%, 50%, 85%, and 100% of light
• Ruler or yard/meter stick or measuring tape (optional)
Observation and Hypothesis
1. Observe the world for a phenomena involving light intensity and photosynthetic
organisms. Describe this phenomenon and ask a question about it.
2. Formulate a hypothesis to explain your observation.
Writing the Procedure
When you develop a protocol for your experiment, it can be helpful to draw a picture of your
experimental setup. Think about the materials you have and the question you are trying to
answer. Illustrate your setup and articulate your procedure. Ask your teacher to review the
experiment before you begin.
Results
Record any observations relevant to your experiment.
Data Analysis and Interpretation
Graph your results
Make any calculations that may be relevant to your data:
What do these calculations/results mean?
How do the data support or contradict your hypothesis?

Revisit your diagram of photosynthesis and cellular respiration from Investigation #1. Revise
your
diagram to demonstrate how the experimental design of your current investigation affects these
processes. Indicate where the rates of these processes may increase or decrease and describe
why. Also indicate how the organism will be affected if the conditions of your investigation were
to continue. List any ideas you have for further refining your hypothesis and testing your
experimental design.

Investigation #4: Effect of Light Color

In nature, light quality, or the color or wavelength of light available to an organism for
photosynthesis, can fluctuate depending on a variety of conditions, including location
and weather. In the laboratory or classroom, color filters, which reduce transmittance of
specific wavelengths of light, can be used to manipulate the quality of light available to your
samples. In this investigation, you will design and conduct an experiment using color filters
to understand the impact of light quality (color) on the rate of photosynthesis by the
algae beads.
For this investigation, all the materials and equipment from Investigation #2 will be made
available to you as well as the following:
• Color filters that allow only specific colors (wavelengths) of light to pass through
Observation and Hypothesis
1. Observe the world for phenomena involving light color and photosynthetic organisms.
Describe one phenomenon and ask a question about it.
2. Formulate a hypothesis to explain your observation.
When you develop a protocol for your experiment, it can be helpful to draw a picture of your
experimental setup. Think about the materials you have and the question you are trying to
answer. Illustrate your setup and articulate your procedure. Ask your teacher to review the
experiment before you begin.
Results
Record any observations relevant to your experiment.
Data Analysis and Interpretation
Graph your results
Make any calculations that may be relevant to your data:
What do these calculations/results mean?
How do the data support or contradict your hypothesis?

Revisit your diagram of photosynthesis and cellular respiration from Investigation #1. Revise
your diagram to demonstrate how the experimental design of your current investigation affects
these processes. Indicate where the rates of these processes may increase or decrease and
describe why. Also indicate how the organism will be affected if the conditions of your
investigation were to continue. List any ideas you have for further refining your hypothesis and
testing your experimental design.

Investigation #5: Effect of Temperature

All living organisms have a wide variety of adaptations that allow them to survive
temperature extremes. For example, some plants have adapted to extreme conditions like
the Arctic tundra. Temperature is another environmental factor that can affect the rate of
photosynthesis, as well as respiration, in the algae beads. In this investigation, you will design
and conduct an experiment to understand the impact of temperature on the rate(s) of one or
both of these Processes. For this investigation, all the materials and equipment from
Investigation #2 will be made
available to you as well as the following:
• Water bath
• Ice
• Thermometer
Observation and Hypothesis
1. Observe the world for phenomena involving light color and photosynthetic organisms.
Describe one phenomenon and ask a question about it.
2. Formulate a hypothesis to explain your observation.
When you develop a protocol for your experiment, it can be helpful to draw a picture of your
experimental setup. Think about the materials you have and the question you are trying to
answer. Illustrate your setup and articulate your procedure. Ask your teacher to review the
experiment before you begin.
Results
Record any observations relevant to your experiment.
Data Analysis and Interpretation
Graph your results
Make any calculations that may be relevant to your data:
What do these calculations/results mean?
How do the data support or contradict your hypothesis?

Revisit your diagram of photosynthesis and cellular respiration from Investigation #1. Revise
your diagram to demonstrate how the experimental design of your current investigation affects
these processes. Indicate where the rates of these processes may increase or decrease and
describe why. Also indicate how the organism will be affected if the conditions of your
investigation were to continue. List any ideas you have for further refining your hypothesis and
testing your experimental design.

Investigation #6: Mini-Ecosystem

In nature, organisms are generally not found in isolation. They are part of a complex
and dynamic ecosystem where organisms interact to exchange energy and matter. In
Investigation #2 you studied the balance between photosynthesis and cellular respiration in
a single photosynthetic organism.
How might you investigate photosynthesis and cellular respiration in an ecosystem?
For this investigation, all the materials and equipment from Investigation #2 will be made
available to you as well as the following:
• Aquatic snails or another heterotroph

Observation and Hypothesis

 1. Observe the world for phenomena involving light color and photosynthetic organisms.
Describe one phenomenon and ask a question about it.
2. Formulate a hypothesis to explain your observation.
When you develop a protocol for your experiment, it can be helpful to draw a picture of your
experimental setup. Think about the materials you have and the question you are trying to
answer. Illustrate your setup and articulate your procedure. Ask your teacher to review the
experiment before you begin.
Results
Record any observations relevant to your experiment.
Data Analysis and Interpretation
Graph your results
Make any calculations that may be relevant to your data:
What do these calculations/results mean?
How do the data support or contradict your hypothesis?

Revisit your diagram of photosynthesis and cellular respiration from Investigation #1. Revise
your diagram to demonstrate how the experimental design of your current investigation affects
these processes. Indicate where the rates of these processes may increase or decrease and
describe why. Also indicate how the organism will be affected if the conditions of your
investigation were to continue. List any ideas you have for further refining your hypothesis and
testing your experimental design.