Banh 1

Athena Banh
AP Biology Pd. 3
Giannou
4/6/2014
I. Title
Transpiration Rates of Plants Under Different Conditions, Visualize Plant Stem Cell
Structure, and Observe the Epidermis of Plant Leaves that are Accustomed to Different
Environments
II. Introduction
a. Problem/Purpose: Study the transpiration of a plant under different environmental
conditions by applying knowledge of diffusion, osmosis, adhesion and water potential, and
stomata concentration.
b. Background Information: Plants may be divided into groups based on their structure
and method of water transportation. The plant that will be researched in this experiment, the
tomatillo plant, is one of the vascular group, meaning that water moves upward through the plant
through the xylem tissue. The phloem tissue also aids water and solute movement in vascular
plants. Vascular plants do not contain a physical pump that transports water, but rather instead
rely on air pressure to push water from the roots, or hydrostatic root pressure, in which osmosis
moves the water into the xylem. The transport and movement of water in a plant is affected by
water potential in which water moves to an area with a higher water potential. Vascular plants
may also rely on the transpiration pull theory, or cohesion, to bring water into it. Transpiration
occurs with a large water loss from the plant by evaporation from the opening of the stomata.
The opening of the stomata causes evaporation due to the higher water potential of the

Banh 2

surrounding air. The overall movement of water through the plant because of cohesion results in
a transpirational pull from the leaves to the roots of a plant. Despite the many ways in which a
plant obtains the solutes and water needed to sustain itself, environmental factors, such as
temperature, light intensity, and air currents, greatly impact the rate of transpiration, such factors
have an effect on the opening and closing of the stomata and evaporation rate. The epidermis of
plants that require more water to sustain itself differ from that of plants that are less dependent of
water for survival.
i. Terms
Vascular Plants- plants that have tissues that are specifically designed for efficient
water and solute transport
Xylem- vascular tissue in plants that conducts water and dissolved nutrients
upward from the root
Phloem- vascular tissue in plants that conducts sugars and other metabolic
products downward from the leaves
Osmosis- process of molecules passing through the plant membrane from a less
concentrated solution to a more concentrated one
Water potential- explains the water movement from cell to cell over long
distances in the plant
Cohesion- process of water being pulled up the xylem from the roots and soil due
to transpiration
Stomata- pores in the epidermis of the leaf or stem of a plant that allow for
exchange of water and air with the environment
Potometer- a tool used to measure the rate of transpiration in a plant by

Banh 3

determining the amount of water absorbed
Parenchyma- plant tissues that are found in roots, stems and leaves
Sclerenchyma- plant tissues that function as support
Collenchyma- fundamental nonspecialized plant tissue
Epidermis- outer covering of roots, stems and leaves that functions as a barrier
to water loss and protection against injury
c. Hypothesis: Because environmental factors affect the opening and closing of the
stomata on the leaf surface, the rate of transpiration will change accordingly.
d. Variables: For Part A (transpiration rate in different environments), the independent
variables are the different environmental factors: air temperature, light intensity, and air current,
and the dependent variable is the rate of transpiration. For Part B (visualizing the plant stem cell
structure), the independent variable is stem cell structure, and the dependent variable is also the
stem cell structure because it is solely an observational procedure. For Part C (observing the
epidermis of the of different plants), the independent variable is the amount of water the plants
require, and the dependent variable is the amount of stomata on the epidermis.
III. Methods
a. Materials
Part A Materials:
-potometer

-tomatillo plant

-water

-ring stand

-.1mL pipet

-16” vinyl tubing

-petroleum jelly

-timer

-100 Watt light bulb -fan

-mister

-marker

-paper towels

-ruler

-razor

-scale

-scissors

Banh 4

Part B Materials:
-microscope

-tomatillo plant

-razor

-nut and bolt microtome

-Pasteur pipet

-parawax

-spatula

-50% ethanol

-Toluidine blue O stain

-distilled water

-50% glycerol

-cover slip

-microscope

-microscope slides

-Impatiens plant

-succulent plant

-microscope

-microscope slides

-cellophane tape

-clear nail polish

-scissors

-petri plate

Part C Materials:

b. Procedures:
Part A Procedures:
1. Place non-tapered end of a .1mL pipet into one end of a 16” piece of vinyl tubing.
Petroleum jelly may be placed on the outside of the pipet to help the insertion of it into the
tubing.
2. Bend the tube into a U-shape and place it onto a ring stand with clamps. The top of the
tubing and the tops of the pipet must be level.
3. Fill the tubing and pipet with water from the tubing end using a .1mL pipet. No air
bubbles should be present.
4. Insert a freshly cut stem from a tomatillo plant into the end of the tubing and seal with
petroleum jelly.
5. Expose the stem to one of the following conditions:
-room conditions
-100 watt light source place 1 meter from the stem
-Fan 1 meter from the stem on low

Banh 5

-mist of water from plant mister, covering the potometer immediately with a
plastic bag
6. After a 10 minute equilibrium period, read the water level of the pipet and record as
time 0. Use a marker to draw a line at the water level for each time point.
7. Obtain a reading once every 5 minutes for 30 minutes.
8. After 30 minutes, cut off the leaves of plant and blot off any excess water. Weigh the
leaves.
9. Estimate the total leave surface area for the plant.
Part B Procedures:
1. Assemble the nut and bolt microtome to where there is a small cup at the end.
2. Cut a slice of the tomatillo stem about 5-6mm in length.
3. Place the cut stem into the microtome so that it stands up. Use a Pasteur pipet to fill the
cup with melted parawax.
4. Wait 5-10 minutes for the parawax to solidify. When it has solidified, lay the
microtome on its side and with a razor, slice away the excess wax.
5. Twist the bolt to expose a thin piece of wax. Slice off a thin section of the wax/stem.
Using a spatula, place the sections in a petri dishes with 50% ethanol. Prepare 10-12 sections.
Let the sections soak for 5 minutes.
6. Transfer the sections to the Toluidine blue O stain solution in another petri plate. Stain
for 5-10 minutes.
7. Transfer the stained sections to the petri plate with distilled water.
8. Place the sections onto a microscope slide and add a drop of 50$ glycerol, and cover
with a cover slip.

Banh 6

9. Observe the sections under a compound light microscope.
10. Draw the observed structures.
Part C Procedures:
1. Obtain a succulent leaf and an Impatiens leaf.
2. Paint a solid patch of clear nail polish on the top and bottom of the leaves of each
plant. Make the patch at least one square centimeter.
3. Allow the nail polish to completely dry.
4. Press a piece of clear cellophane tape onto the dried nail polish patch.
5. Peel the tape away carefully.
6. Tape the peeled impression to a microscope slide.
7. Examine the leaf impressions under a light microscope.
8. Observe and draw the structures, including the stomata.
IV. Data and Calulations
a. Data Tables:
Potometer Readings (Room Conditions)
Time (min)
Reading (mL)

0

5

10

15

20

25

30

.009

.0125

.0167

.0192

.0225

.0258

.0283

Individual Water Loss in mL/m² (Room Conditions)
Time (min)

0

5

10

15

20

25

30

Water Loss (mL)

.009

.0125

.0167

.0192

.0225

.0258

.0283

Water Loss per m²

3.33

1.29

2.85

3.78

5

6.22

7.15

Banh 7

Class Average in Water Loss in mL/m2
Time (min)
Treatment 0

5

10

15

20

25

30

Room

2.79

1.71

2.80

3.64

4.6

5.74

6.58

Light

3.62

.878

1.82

2.58

3.08

3.95

4.46

Fan

1.49

1.51

2.66

3.37

4.18

4.81

5.56

Mist

1

-.88

-.75

1

1

1

1

b. Graphs
Individual Potometer Readings (Room Conditions)
0.03

Reading (mL)

0.025
0.02
0.015
0.01
0.005
0
0

5

10

15

20

25

30

35

Time (min)

Individual Water Loss (Room Conditions)

Water Loss per m²

8
6
4
2
0
0

5

10

15

20

Time (min)

25

30

35

Banh 8

Class Average Water Loss
7
6

Water Loss (mL/m²)

5
4

Series1

3

Series2

2

Series3

1

Series4

0
-1

0

5

10

15

20

25

30

35

-2

Time (min)

c. Calculations:
Total Leaf Surface Area:
Water Loss:

=

= .0027 m²
=

= 1.29 mL/m²

V. Conclusion and Evaluation
a. Concluding: As seen in the results of the differing environmental conditions of Part
A’s procedures, the plant that was exposed to the classroom conditions experienced the greatest
loss of water. Factors that contributed to the water loss of the plants in the room conditions were
the temperature, air flow, and exposure to light. This plant probably showed the most dramatic
loss in water because it was exposed to two of the other three conditions, both of which caused a
loss individually as well. The plant that was exposed to the mist conditions retained the most
water throughout the experiment as the humidity in the plastic bag kept water from escaping both
the plant and its environment. Part C’s procedures of observing the stomata density of plants that
require different water levels to survive revealed that the conditions of a plant’s natural habitat

Banh 9

affect transpiration rate. The Impatiens plant, one that requires a sufficient amount of water, had
a significantly higher stomata density on both the upper and lower epidermis when compared to
that of the succulent, a cacti, thus requiring less water.
b. Evaluating Procedures: Part A of the experiment had very clear and detailed
instructions and diagrams that displayed the intentions of the experiment thoroughly and were
easy to follow. Part B proved to be slightly more difficult to follow as the diagrams only showed
one step of the process, which seemed tedious and difficult at times. Though I did not physically
do Part B except for observing the end product through the microscope, my group members
seemed confused and lost at times. Part C’s procedures were clearly numbered and listed and the
procedure was easy to follow overall as its minimal tasks were very simple.
c. Improving the Investigation: The investigation could have been improved had we had
more time to complete the experiment so that each individual could partake in all three of the
procedures so that we could more fully understand the objective of each procedure. Both Part A
and Part C had instructions that were organized logically and easy to follow. Part B could be
improved by providing clearer instructions and more diagrams to demonstrate what was
supposed to be achieved per number instruction.
VI. Questions:
1. How can you calculate the total leaf surface area expressed in cm²? In m²?
The total leaf surface area in m² can be found through weighing all of the leaves
in grams and then separately weighing a 1 cm² section of the leaf in grams.
Multiply the cut section mass by 10,000 to get the mass per square meter of the
leaf and then divide the total weight by the mass per square meter to get the total
surface area.

Banh 10

2. How can you estimate the leaf surface area of the entire plant without measuring every
leaf?
The leaf surface area of the entire plant can be measured without measuring every
leaf by finding that of one leaf and then estimating the amount of leaves on the
plant, and then multiplying the single leaf surface are by the estimation of the
total amount of leaves.
3. What predictions and/or hypotheses can you make about the number of stomata per
mm² and the rate of transpiration?
The greater the number of stomata that can be observed through a microscope, the
greater the transpiration rate of the plant.
4. Is the leaf surface area directly related to the rate of transpiration?
Yes, the greater the leaf surface area, the greater the rate of respiration.
5. What predictions can you make about the rate of transpiration in plants with smaller or
fewer leaves?
The rate of transpiration in plants with smaller or fewer leaves is less than that of
a plant with bigger or more leaves.
6. Because most leaves have two sides, do you think you have to double your calculation
to obtain the surface area of one leaf? Why or why not?
No, because surface area is only in regards to one side of the leaf.
7. Water is transpired through the stomata, but carbon dioxide must pass through the
stomata into a leaf for photosynthesis to occur. There is evidence that the level of carbon
dioxide in the atmosphere had not always been the same over the history of life on Earth.

Banh 11

Explain how the presence of a higher or lower concentration of atmospheric carbon
dioxide would impact the evolution of stomata density in plants.
A higher concentration of atmospheric carbon dioxide would result in plants
having a lower stomata density. The reverse of a lower concentration of
atmospheric carbon dioxide would result in plants with a higher density of
stomata.
8. Based on the data in the following table, is there a relationship between the habitat (in
terms of moisture) to which the plants are adapted and the density of stomata in their
leaves? What evidence from the data supports your answer?
The plants with a greater stomata density in its lower epidermis most likely live in
a warmer, more humid region, as the stomata on the bottom of the leaf would
butter prevent water of evaporating. The plants with a greater stomata density in
its upper epidermis are most likely in a cooler region where water is less likely to
evaporate from the plant. As seen in the table, most plants have greater stomata
densities in their lower epidermis compared to their upper epidermis which
reflects on the fact that having the stomata in the lower epidermis is more reliable
in terms of water and carbon dioxide maintenance in the plant.
VII. Literature Citation;
ED VOTEK: Principles of Transpiration. Washington, DC: ED VOTEK, 2014: Pages 4-14.
Print.