Laraib Awan

AP Biology

Schulte

Experiment # 4

Date of Experiment: 2/14/18

Measuring and calculating the rate of oxygen consumption in germinating seeds using
respirometry techniques
 Abstract
The purpose of this lab was to measure the rate of oxygen consumption, also known as
cellular respiration, in germinating mung beans which were compared to non germinated mung
beans and glass beads in an ice bath around 10 degrees celsius. It is predicted that if germinated
seeds are present, then the rate of oxygen consumption will increase as the hypothesis.
Three respirometers were used throughout the duration of this lab: one with 50 mL of
water and 2 mL of mung beans, one with 50 mL of water and 25 non germinated mung beans to
equal 53 mL, and one with 50 mL and 34 glass beads in order to equal it to 53 mL. The
respirometers were then drained and the solid material which was placed in these paper towels
were taken out and put on a paper towel to dry. Absorbent cotton, which was drenched in 15%
KOH to activate the process of cellular respiration, and non absorbent cotton to block any air
flow. Because germinated plants undergo cellular respiration more than non-germinated seeds, it
is reasonable to assume that often the germinated mung beans are more likely to consume more
oxygen than the other solid materials. Masking tape was used to stop airflow from entering the
respirometer because if air were to enter the respirometer, the experiment would fail as carbon
dioxide would come from other human sources and not solely the seeds and beads. It was to be
tested in two different temperatures of water; however, the experiment was only conducted in 10
degrees celsius. Therefore, the temperature was not measured and the only things compared was
the measurement of oxygen which was consumed.
After the lab was conducted, it was observed that the germinated seeds did consume more
oxygen at 4.75 mL while the beads consumed the least amount of oxygen at 2.5 mL. The
hypothesis was proven to be correct as the germinated seeds did consume the most oxygen.

Materials
● 50 germinating pea seeds
● 50 dry seeds
● 100 plastic beads
● 3 respirometer vials
● Weights for vials
● 3 stoppers
● 1 mL graduated pipette
● Sealant
● 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 of clock
● Water
● Dropper bottle of 15% KOH
Procedures
1. A 10 degree celsius bath should be set up immediately to allow time to adjust the
temperature.
2. Respirometer 1: Obtain a 100-mL graduated cylinder and fil lit with 50 mL of water.
Place 25 germinating peas into the graduated cylinder. Measure the amount of water that
was displaced and record this number. Mung bean volume: _2 mL_
3. Respirometer 2: Refill the graduated cylinder until it has 50 mL of water. Place 25 dried
mung means into the graduated cylinder and then add enough glass beads to reach a
volume equivalent to the volume of the germinating peas in Respirometer 1. Remove
these beans and beads and place them on a paper towel. These peas 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 peas 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 that 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 set of germinating peas, dry peas + beads, and beads in vials one, two, and
three, respectively.
7. Insert the non tapered end of one pipet into the upper surface of one of the rubber
stoppers. It should fit tightly. Place a layer of sealant around the junction between the
pipet and the stopper so that no air can escape. (*The pointy end of the pipet should be
outside, not inside)
8. Wrap parafilm tightly around each stopper (where it enters 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
2/3 of the way from one end.
 10. Place all three respirometers in the ice bath and begin timer for 7 minutes.
11. At the end of 7 minutes, submerge each of the tubes entirely in the water bath. Some
water will enter the tip of the pipet, but the influx of water should stop fairly quickly. If it
does not stop, check the respirometer 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 pipet. The pipet 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 here, stainless steel
dissection scissors; for instance, can serve to weight the tubes.
13. Record the water level in each pipet onto the data table at the Time Interval 0. 6. Record
the position of the water in each pipet at the end of 5, 10, and 15, 20 min on Data Table 2.

Results

Respirometer Temperature Contents

1 10 degree celsius Germinated seeds

2 10 degree celsius Dry seeds + Beads

3 10 degree celsius Beads

This is a graph which displays what was put in each respirometer at what temperature in the ice
bath.
Respirometer 1 Respirometer 2 (non Respirometer 3
(germinating seeds) germinating) (beads only)

Initial Volume (mL) 50 mL 50 mL 50mL

Final Volume (mL) 53mL 53 mL 53mL

Volume of 3 mL of seeds 2 mL of seeds (25) 3 mL of beads (34)

beads/seeds 1 mL of beads (17)
This chart shows the initial and final volumes along with the volumes of the beads/seeds which
were subtracted from the equation. The amount of seeds and beads required are also shown.
Final Volumes
Germinating seeds volume Non germinating seeds Beads volume
volume

4.75 mL 4.5 mL 2.5 mL

This chart exhibits the final volumes which were discovered at the end of the experiment in the
respirometer.
 Analysis
The purpose of this lab was to calculate and observe which form of seeds consume the
most oxygen. The germinated seeds turned out to consume the most oxygen as it had produced a
volume of 4.75 mL when put in an ice bath with a temperature of 10 degrees celsius. Beads,
which are an abiotic factor, exhibited signs of oxygen consumption as it obtained a volume of 2.5
mL; however, because beads are unable to undergo the process of cellular respiration, the
volume should have been 0 mL. This may have been caused due to human error in the lab. The
non germinating seeds produced a volume of 4.5 mL which was fairly close to the volume that
the germinating seeds had produced; however, no cellular respiration should have occurred with
the non-germinated seeds since they do not produce nor receive to gases needed for cellular
respiration to occur (oxygen, carbon dioxide). This .25mL difference may have been caused due
to human error.
The hypothesis that if germinated seeds were present, then the rate of oxygen
consumption would increase was proven to be correct; however, the experiment underwent
several errors caused by a human source which could have skewed the results and cause this to
be incorrect. The non-germinated seeds and glass beads were expected to not produce any
carbon dioxide as measured by the volume in the respirometer but the beads produced little
carbon dioxide of 2.5 mL and the non-germinated seeds had produced similar amounts of carbon
dioxide as the germinated seeds with only a .25 mL difference.
There were several errors with the source of humans throughout this lab. It is possible
that air had entered the respirometers, allowing other sources to produce carbon dioxide. It is
also possible that different seeds might have gone into their unassigned respirometers, leading in
flaws of the results.
The hypothesis was proven correct; however, to get accurate results, this experiment
should be done multiple times to get rid of human error.

Background research

It is important to see how different temperatures affect the process of cellular respiration
and the rate of oxygen consumption. Enzymes play a significant role in the process of cellular
respiration and when these enzymes are denatured, it interrupts the process. Testing this
experiment in several temperatures will give researchers a broad idea of how well cellular
respiration works in different temperatures and towards what temperature the enzymes become
denatured.
According to the St. Vincent College, cellular respiration works better in higher
temperatures; however, once the temperature increases or decreases towards the extremities, the
enzyme will denature and disrupt the process of cellular respiration.
 Citations

Campbell, N. A., Urry, L. A., Cain, M. L., Wasserman, S. A., Minorsky, P. V., & Reece,
J. B. (2017). ​Campbell biology in focus​. Harlow, Essex, England: Pearson Education Limited.

S. V. (n.d.). Does temperature change affect the respiration of yeast. Retrieved March 1,
2018, from
https://vle.stvincent.ac.uk/2014/pluginfile.php/33446/mod_resource/content/2/Does-temperature-
change-affect-the-respiration-of-yeast-10vsrbd%20(1).pdf