Lauren Dattilo

Ms. Schulte

Experiment: #5

February 14, 2018

"The effect germinating versus non-germinating seeds on oxygen consumption during

respiration"

Abstract
 Cells break down nutrients molecules in order to generate ATP which is then used to for
cellular process that require energy. This process overall can be defined as cellular respiration.
Cellular respiration uses nutrient molecules (glucose) and oxygen in return provide carbon
dioxide and a waste product of water. There are several methods used to measure this process.
Some examples would be to measure the heat produced, oxygen consumption, and even the
carbon dioxide production.
In this lab you will use respirometers in order to measure the gas volume changes in
return giving you data on the rate of cellular respiration. Since there are two gases involved in
this processes KOH is introduced. KOH reacts with carbon dioxide to create a solid precipitate.
This allows for only the oxygen gas to change pressure within the respirometer. When the
oxygen is consumed the overall pressure in the respirometer will decrease, causing the water to
move away from the air with low pressure.
Using this process with the respirometers, you will compare the different oxygen
consumption levels of germinating seeds and non germinating seeds. You will also use a
respirometer with only glass beads as a control that will also allow you to see any atmospheric
pressure changes.

Materials and Equipment Used

○ 50 germinating seeds
○ 50 dry seeds
○ 100 plastic beads
○ 3 respirometer vials
○ Weights for vials
○ 3 stoppers
○ 1 mL graduated pipettes
○ Sealant (Vaseline)
○ 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 or clock
○ Water
○ Dropper bottle of 15% KOH
Procedures
*note: it is important to remember that you must grow the germinated seeds before performing
this lab
Day One
 1. Build a room-temperature bath (approximately 25 degrees celsius) and a 10 degree
celsius bath should be set up immediately to allow for time to adjust the temperature of
each. (Add ice to attain the 10 degree celsius bath)
2. Respirometer 1: Obtain a 100-mL graduated cylinder and fill it with 50 mL of water.
Place 25 germinating peas 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 peas and will be used to determine the volume of the
peas and beads in the other respirometers (all volumes should be equal). Remove the peas
and place them on a paper towel. These peas will be used in Respirometer 1.
3. Respirometer 2: Refill the graduated cylinder until it has 50 mL of water. Place 25 dried
peas (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 peas in
Respirometer One. Remove the peas 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. Repeat the above procedures (2-4) to prepare a second set of germinating peas, dry peas
and beads, and beads for use in Respirometers 4, 5, and 6, respectively.
6. Your group will be assigned to do one of the temperature sets.
7. To assemble the respirometers, obtain 3 vials, each with an attached stopper and pipette.
Make sure 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, staurate 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.
8. Place the first set of germinating peas, dry peas and beads, and beads in vials one, two
and three, respectively.
9. Place the second set of germinating peas, dry peas and beads, and beads in vials four,
five, and six, respectively.
10. 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.
11. 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. Set your apparatus aside for day 2.
Day Two
1. Place a strip of masking tape across the narrow width of the water bath, approximately ⅔
of the way from one end.
2. Place respirometers 1, 2, and 3 into the room temperature water bath and respirometers 4,
5, and 6 into the cold water so that the pipettes rest on the masking tape prop. Begin time
for a total of 7 minutes (this is the equilibration 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.
 3. 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.
4. 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 here, stainless steel
dissection scissors; for instance, can serve to weight the tubes.
5. Record the water level in each pipette onto the data table at the Time interval 0.
6. Record the position of the water in each pipette at the end of 5, 10, and 15, 20 minutes.

Results

Cellular Respiration Germinating Seeds Non-Germinating Seeds

Room Temp Time Reading Corrected Reading Corrected

27 degrees 0 0 0 0 0
Celsius
5 0 0 0 0

10 0 0 0 0

15 .76 mL .76 mL .59 mL .59 mL

20 .84 mL .84 mL .62 mL .62 mL

Based on the data in the above table, you are able to see that more oxygen was consumed by the
germinating seeds. The germinating seeds consumed about ⅔ the oxygen of the non-germinating
seeds at room temperature.

Cellular Respiration Germinating Seeds Non-Germinating Seeds

Cold Time Reading Corrected Reading Corrected

4 degrees 0 0 0 0 0
 Celsius 5 0 0 0 0

10 0 0 0 0

15 .71 mL .71 mL .75 mL .75 mL

20 .77 mL .77 mL .78 mL .78 mL

In this table you can see that both germinating and non-germinating seeds consumed around the
same amount of oxygen at a cold temperature. In specific the same amount of water was
observed in cold temperatures between the two types of seeds.

Analysis/Conclusion
Based on the tables and information obtained with this experiment, we can conclude that
the use of respirometers are a valid unit to measure the rate of cellular respiration with seeds. At
room temperature we also observed that the amount of water collected in the pipette was greater
for that of germinating seeds then it was of the non-germinating.This means that there was more
oxygen consumed in the respirometer containing germinating seeds because there was a greater
change in pressure within the device causing for more water to be recorded. This data proves that
when it comes to cellular respiration at room temperature germinating seeds are more efficient
and consume more oxygen. However when placed in the water bath, Both seeds had the same
amount of water recorded in the pipette. Meaning both germinating and non germinating seeds
consume the same amount of oxygen at colder temperatures.
For further experiment different organisms can be tested within the respirometers. For
example, this lab could be conducted using a cricket or a worm instead of seeds. It would be
interesting to compare the date obtained between the two organisms. It would also be interesting
to test the difference considering the cells are different and to see if one cell (animal or plant) is
more efficient at the process then the other.