Vivien Coombs

AP Biology Period 4
Schulte
Experiment #5
February 14, 2018

The Measurement of Cellular Respiration through the Oxygen

Consumption in Both Germinating and Nongerminating Seeds

Abstract
 This experiment shows the rate of cellular respiration that occurs in germinating
seeds, non-germinating seeds, and plastic beads through the amount of oxygen that is
consumed. We conducted this experiment using seeds that had spent about a week
germinating; this was a crucial part of the experiment since, if the seeds were not
germinated, the experiment would not have worked. The plastic beads served as the
control group since the plastic would not show signs of cellular respiration. The results
came out as expected for both the germinating and non-germinating seeds, with the
germinating seeds consuming more oxygen than the non-germinating cells; although
there was supposed to be no oxygen consumed at all in the plastic beads, there
showed that there was some at the 20 minute mark, this means that human error came
into play and did not display an accurate reading. I believe that the rate of oxygen
consumed in the germinating seeds should have been a lot higher than it was but due to
human error it did not perform as fast as it actually would have. The purpose of this lab
was to determine how important the germination process is to oxygen consumption in
cellular respiration; This experiment was supposed to prove that the germinated seeds
would consume more oxygen thus meaning that they performed a higher rate of cellular
respiration; the non germinated seeds would perform cellular respiration but not at quite
as fast a rate; then the plastic beads would not perform cellular respiration due to them
being inanimate objects that do not possess life.

Materials

● 50 germinating pea seeds

● 50 dry seeds
● 100 plastic beads
● 3 respirometer vials
● 3 stoppers
● 1mL graduated pipettes
● Absorbent cotton
● Non-absorbent cotton
● 1 round wood stick
● 3 pieces of paper towel
● Marking pen
● Water bath
● Ice
● 100 mL graduated cylinder
● Thermometer
● Masking tape
● Stopwatch
● Water
● Dropper bottle of 15% KOH
●
Procedure
1. Set up a cold bath; fill a tub with water and ice to get the temperature down to
10oC.
2. Respirometer 1: Obtain a 100 mL graduated cylinder and fill it with 50 mL of
water. Place 25 germinating seeds* into the graduated cylinder. Measure the
amount of water that was displaced (which is equivalent to the volume of the
peas) and record this number. This number is the volume of the seeds, and will
be used to determine the volume of seeds and beads in the other respirometers
(all volumes should be equal). Remove the peas and place them on a paper
towel. These seeds will be used in Respirometer 1.
3. Respirometer 2: Refill the graduated cylinder until it has 50 mL of water. Place 25
dried seeds (they are dormant, and not germinating) into the graduated cylinder
and then add enough glass beads to reach a volume equivalent to that of the
germinating seeds in Respirometer 1. Remove the seeds and beads and place
them on a paper towel. These seeds and beads will be used in Respirometer 2.
4. Respirometer 3: Refill the graduated cylinder until it has 50 mL of water. Fill it
with glass beads alone until the volume is equivalent to the volume of the
germinating seeds in Respirometer 1. Remove these beads and place them on a
paper towel. These beads will be used in Respirometer 3.
5. To assemble the respirometers, obtain 3 vials, each with an attached stopper
and pipette. Make sure that the vials are dry on the inside. It is important the
amounts of cotton and KOH be the same for each respirometer. Place a
small wad of absorbent cotton in the bottom of each vial and, using a dropper,
saturate the cotton with 15% KOH. Do not get KOH on the sides of the
respirometer. Place a small wad of dry non-absorbent cotton on top of the KOH-
soaked absorbent cotton.
6. Place the first set of germinated seeds, dry seeds and beads, and beads in vials
1,2,and 3 respectively.
7. Insert the non-tapered end of one pipette into the upper surface of one of the
rubber stoppers. It should fit tightly. Place a layer of sealant around the junction
between the pipette and the stopper so that no air can escape. (The pointy end
of the pipette should be outside, not inside)
8. Wrap parafilm tightly around each stopper (where it enters the the vial and where
the pipette is inserted). This is necessary to ensure against any leaks. Any leaks
will cause the experiment to fail.
9. Place a strip of masking tape across the narrow width of the water bath,
approximately ⅔ of the way from one end.
 10. Place respirometers 1, 2, and 3 into the cold bath so that the pipettes rest on the
masking tape prop. Begin time for a total of 7 minutes - this is the equilibrium
period, where your respirometers will become the same temperature as the
water. Use a thermometer to determine the water temperature of each bath and
record the measurement.
11. At the end of 7 minutes, submerge each of the tubes entirely in the water bath.
Some of the water will enter the tip of the pipette, but the influx of water should
stop fairly quickly. If not, check for leaks.
12. At this point, check to make sure you can read the pipettes. The air bubble
should extend from the main chamber up the tube of the pipette. The pipette may
need to be rotated so that you can see the numbers. If your respirometers float,
you may need to weight them. Some come with weights inside and some do not.
You can improvise form here and add whatever works.
13. Record the position of the water in each pipette at the end of 10 and 20 minutes
on the data table.

*Prior to this experiment you need to have already germinated the seeds for a least 4-5
days, preferably a week, or else this experiment will not work.
KOH is a chemical that will burn when in contact with skin or eyes; use latex glove when
working with the chemical and use caution.
The independent variables are the seeds, both germinating and non-germinating, and
the dependent variable is the rate of cellular respiration measured through oxygen
consumption.
Results

mL of Water at 10 mL of Water at 20
Minutes Minutes

Respirometer #1 3.25 mL 4.75 mL

(germinating seeds)

Respirometer #2 (Plastic 3.00 mL 4.5 mL

beads and non-
germinating seeds)

Respirometer #3 (only 0 mL 2.5 mL

plastic beads)

This table records the amount of water that has travelled up the respirometers at both
10 minutes and 20 minutes in all three respirometers.

Analysis
 The germinated seeds used had been germinating for about 7 days total when
we used them for this experiment which proved to be a good time frame fro the seeds to
spend germinating. I believe that the rest of the experiment worked well and was able to
express the purpose of the experiment; however, there were some human errors that
occured during our experiment. The stoppers were not secured enough and most likely
had some leaks, hence the amount of water that ended up in the respirometer with the
glass beads despite them not being able to carry out cellular respiration. The
respirometer with the germinating seeds should have resulted in a higher rate of oxygen
consumption because the seeds were already growing and had gotten used to the
process of releasing oxygen, whereas the non germinated seeds had to start from
scratch and just start beginning the process; the cells in the non-germinated seeds have
not had time to aquire ATP and begin the cellular process. Also the experiment is not as
accurate an example due to the water bath being one of 10oC; the cold of the water
slowed down the rate of the oxygen consumption because the enzymes were not able
to work as fast. If the water would have been one of room temperature than the data
would have shown an great increase in the rate of oxygen consumed since the
enzymes were in their range of tolerance. Our hypothesis that the germinating seeds
would show the most oxygen consumed was proven; however it was surprising that the
non-germinated seeds consumed as much oxygen as they did, being that they were
very close to the same amounts as the germinated seeds. The most reasonable
explanation for this would be that another human error occured and perhaps that the
seeds had not germinated for as long as they needed and they would have consumed
more oxygen if they had been germinating for two weeks instead of one.

Background
Cellualr respiration is a process in which cells break down nutrient molecules to
generate ATP and exchange energy; cellular respiration can be measured through
oxygen consumption or change in temperature from production of heat. The
consumption of oxygen and ATP synthesis happen in the mitochondria of the cell which
will then use the oxygen to help run other processes in the cell. The respirometers
measure gas volume changes thus determining the rate of cellular respiration; the gas
for the KOH absorbs the carbon dioxide and cause the consumption of oxygen to occur.
When the respirometers are submerged in the water, the water will flow from an area of
high pressure to an area of low pressure which give us the ability too measure the
oxygen being consumed.
Andersen, Paul. “AP Biology Lab 5: Cellular Respiration.” YouTube, 14 Mar.
2012, youtu.be/r9o_VdToClE.
“Cellular Respiration.” Open Learning Initiative: Register for a Course,
oli.cmu.edu/jcourse/workbook/activity/page?context=434a5e6180020ca60197abf697aff
3e4.

“Bioenergetics: Open Access.” Click Here, www.omicsonline.org/scholarly/cellular-

respiration-journals-articles-ppts-list.php.