Biology Transpiration Experiment

Research Question:
How does the light intensity directed towards a plant (room light, 25cm, 50cm and no
light) affect the rate of transpiration in its leaves?

Background information:
The amount of water needed daily by plants for the growth and maintenance of tissues is
small in comparison to the amount that is lost through the process of transpiration (the
evaporation of water from the plant surface). If this water is not replaced by water
uptake, the plant will wilt and may die due to inhibition of photosynthesis.
The transport of water up from the roots in the xylem is governed by differences in water
potential (the potential energy of water molecules). These differences account for water
movement from cell to cell and over long distances in the plant. Gravity, pressure, and
solute concentration all contribute to water potential, and water always moves from an
area of high water potential to an area of low water potential. The movement itself is
facilitated by osmosis, root pressure and adhesion and cohesion of water molecules.

The Overall Process: Minerals actively transported into the root accumulate in the xylem,
increasing solute concentration and decreasing water potential. Water moves in by
osmosis. As water enters the xylem, it forces fluid up the xylem due to hydrostatic root
pressure. But this pressure can only move fluid a short distance. The most significant
force moving the water and dissolved minerals in the xylem is an upward pull as a result
of transpiration, which creates tension. The “pull” on the water from transpiration results
from the cohesion and adhesion of water molecules (although scientists are now also
discovering active “pumps” powered by ATP that help the fluids move against gravity).

Transpiration begins with evaporation of water through the stoma (stomata), small
openings in the leaf surface which open into air spaces that surround the mesophyll cells
of the leaf. The moist air in these spaces has a higher water potential than the outside air,
and water tends to evaporate from the leaf surface (moving from an area of high water
potential to an area of lower water potential). This transpiration pull occurs because of
(1) the cohesion of water molecules to one another due to hydrogen bond formation, and
(2) by adhesion of water molecules to the walls of the xylem cells, which aids in
offsetting the downward pull of gravity.

Evaporation through the open stoma is a major route of water loss in plants. However,
the stoma must open to allow the entry of CO2 and the loss of water by regulating the
opening and closing of stoma on the leaf surface. Many environmental conditions
influence the opening and closing of stoma and also affect the rate of transpiration.
Temperature, light intensity, air currents, and humidity are some of these factors.
Different plants also vary in the rate of transpiration and in the regulation of stomatal
openings.

The plant kingdom can be divided into two major groups: bryophyte and tracheophyta.
The major distinction between these two groups is the presence of specialized vascular
tissue for the transport of water and food. While bryophytes, such as moss, lack
conducting tissues, tracheophtyes utilize specialized cells, xylem and phloem, for the
transport of water and food. The evolution of vascular tissues and the subsequent ability
to transport water and food over greater distances has contributed to the evolution of
larger plants with specialized ground tissues and organs: roots for support and uptake of
water and minerals, stems for support of leaves and flowers, and leaves for
photosynthesis. These adaptations have allowed tracheophtyes to become the dominant
form of terrestrial plant.

Ground tissues, which comprise the plant body, or cortex, are made up of three types of
cells: collenchyma, parenchyma, and sclerenchyma cells. Collenchyma cells are
irregularly shaped, with thick cell walls. They are found in a plant stem, where they help
support the body of a plant. Parenchyma cells are the photosynthetic cells of a plant, with
thin, many-0sided walls; they make up the ground tissue of a leaf. Sclerenchyma cells
are elongated, with primary and secondary walls, containing lignin, a protein that
strengthens the cell. Unlike parenchyma and collenchyma cells, they die at maturity.
Sclerenchyma cells are often found near vascular tissue.

According to the generally accepted cohesion-tension theory, water is pulled up to the

leaves of a plant by transpirational pull. When stomata are open, water transpires from
higher water potential in the mesophyll spaces to lower water potential in the air.
Decreasing water potential in the air spaces pulls water from nearby mesophyll cells,
which in turn pull water from xylem vessels in nearby veins of the leaf.

Due to the cohesive nature of water molecules, when one water molecule is pulled from
the xylem, more and more follow close behind in a chain of water molecules pulled
upward from the roots to the leaves. The tension, or negative pressure, caused by the
upward pull of the water column is so strong that the diameter of a stem actually
decreases when the rate of transpirational pull is very high.

In the root, minerals actively absorbed from the soil increase the solute concentration of
cortical cells. This causes water to flow by osmosis from the soil into the root, creating
hydrostatic pressure known as root pressure. Water from the cortex of the root
continually moves toward the xylem, aided by the push of root pressure, where it is then
pulled up by transpiration. Root pressure and transpiration pull together provide more
than enough force to offset the pull of gravity and raise water to the top of even the tallest
redwood trees.

Although transpiration is the driving force behind water transport, and the sun’s energy is
the driving force for transpiration, plants have evolved many adaptations to prevent
excessive loss of water by transpiration. As expected, the rate of transpiration varies
directly with the amount of sunlight, heat, and wind in the environment, and a delicate
balance must be maintained between stomata closing to prevent water loss and stomata
opening for the exchange of oxygen and carbon dioxide. Structures called guard cells
regulate the opening and closing of the stomata. For instance, in desert plants, guard cells
often keep the stomata closed during the day when the rate of transpiration is high and
open them at night for respiration. Stomata are sensitive to light quality as well as
quantity. They will open in response to energy from the light spectrum. In addition, the
presence of a waxy cuticle on the upper surface of leaves, or small hairs on the lower
surface of leaves, prevents the loss of water. The rate of transpiration also varies
indirectly with humidity in the environment. Cacti, which live in hot arid environments,
have evolved small needle-like leaves to reduce the surface area from which transpiration
can occur; whereas tropical plants in humid and shady environments can afford to have
very large leaves.
Hypothesis:

I think that the closer the lamp is to the plant, the greater the light intensity will be hence
causing the rate of transpiration to be much higher as it causes the stomata to open wider.
In the dark room there will be no transpiration as the stomata will be closed.

Materials and Apparatus:

 Potometer
 Retort stand and clamp
 Vaseline
 Plant with undamaged leaves
 Metre rule
 Table lamp
 Stopwatch
 Scissors
 Large tub/basin
 Rubber bung
 Syringe
 Paper towel

Variables:

Variable Value(s) Justification

Independent Light Intensity Room light, 25cm, To find out the effect of
50cm, dark room different light intensities on the
amount of water transpired.
Dependent Distance the air In cm This will be used to denote the
bubble has moved in amount of water lost from the
6 minutes plant.
Controlled Room temperature 26ºC To ensure that change in
temperature does not affect the
rate of transpiration.
Wind speed constant To ensure that wind speed does
not affect the rate of
transpiration
Size of plant Medium sized To ensure transpiration will not
take too long and make sure all
the readings are accurate.
 Species of plant used
Time interval
between each reading
So that rate of transpiration is
constant for each reading.
6 minutes So the water transpired is
measured at exact intervals
enabling an accurate reading
for each light intensity.

Procedure:

1) Obtain a plastic tub and fill it full of water.

VERY IMPORTANT: During the following steps, the cut end of the plant must always be
under water and not exposed to air. If the end comes out of the water, a small air bubble
will be trapped in the end and the plant will not be able to draw up water as efficiently. If
this happens, you must cut off the end again about an inch up the stem.

2) Rinse the dirt off the roots in the bucket indicated by your instructor.

3) Place the roots in the tub of water.

4) While keeping the roots and lower stem of the plant under water in the basin, cut off
the stem at an angle (Fig. 1), approximately 1 inch from the roots.

Fig. 1. Diagram of how to cut the plant stem.

LEAVE THE CUT END OF THE PLANT UNDER WATER!

5) The leaf’s shoot is placed upright in the potometer’s capillary tube, through a rubber
bung.

6) Quickly smear a layer of lubricant around the area where the plant’s stem and the
tubing come together. This procedure will produce a watertight seal. Dry the leaves using
tissue paper.
7) To trap an air bubble, apply gentle pressure to the syringe.

8) Read the level of the air bubble at time zero and continue recording readings every 6
minutes.

Safety Precautions:
 Be careful while moving photometer from one place to another.
 Wear safety goggles
 Avoid parallax error while taking readings.
 Take care not to leave any air bubbles while placing plant into photometer.
 Be cautious while cutting the plant shoot

Raw data:
Distance travelled by air bubble in 6 minutes (in cm)
Light intensity at: Initial Final
Room light 0.8 1.2
50cm 1.7 2.0
25cm 2.0 2.9
Dark room 2.9 3.0

Processed data:
Calculations:
Rate of transpiration= distance travelled by bubble(cm)
Time taken for bubble to move that distance (s)

Room light:
(1.2-0.8) = 0.001111 cm/s
(6*60)

50cm:
0.3 = 0.000833 cm/s
360

25cm:
0.9 = 0.002500 cm/s
360

Dark room:
0.1 = 0.000278 cm/s
360
Table showing the rate of transpiration at each light intensity:
Light intensity Rate of transpiration (cm/s) ±0.0000005
Room light (control) 0.001111
50cm 0.000833
25cm 0.002500
Dark room (0cm) 0.000278

Observations:
As the light intensity got stronger it was easier to view the movement of the air bubble as
compared to room lighting.

Graph of rate of distance of light against rate of transpiration:

0
Rate of transpiration/cms-1

0
Dark room (0cm) 50cm 25cm

Distance at which light was placed/cm

Analysis of graph:

As can be seen from the graph the general trend shows that as the distance is decreased,
therefore as light intensity increases the rate of transpiration also increases. The graph
also shows the large rise in rate of transpiration from 0.000278 cm/s to 0.000833 cm/s as
light was introduced as compared to a dark room with no light. The rate of transpiration
increases three-fold for half the distance, from 0.000833 cm/s at 50cm to 0.002500 cm/s
at 25cm; this shows the drastic effect that light intensity has on the rate of transpiration.

Conclusion:
From the experiment and data collected it can be concluded that there is a definite
relationship between light intensity and the rate of transpiration. Supported by the data it
can be said that the greater the light intensity, the more the distance travelled by the air
bubble and hence the greater the rate of transpiration. From the data we see that the
results where the light was placed at a distance of 50cm and 25cm were much larger than
those in the room light and dark room showing that light is one of the major factors that
effects transpiration. These results prove my hypothesis to be right as there was an
increase in the rate of transpiration as light intensity was increased. However, in the dark
room transpiration did occur which could be as a result of external factors.

Evaluation:
Strengths:
The experiment was conducted in fair conditions and the controlled variables were kept
constant throughout so I feel as though the experiment yielded quite accurate results. The
constant decimal point reading for the distance travelled also allowed for more precise
readings and therefore a more precise calculation for the rate of transpiration. The plant
used had large undamaged leaves which meant that the rate of transpiration was uniform
and no external factors were affecting it.

Limitations:
One of the biggest limitations of this experiment was the amount of trials conducted,
which was only one, this left more room for systematic and random errors which could
not be picked out as there were no other values for comparison. This greatly reduced the
accuracy and reliability of the results. Another limitation was the use of a phone light to
view the distance travelled by the air bubble in the dark room which may have interfered
with the results, causing transpiration to occur. The readings could be taken to more
decimal places to further increase accuracy. There were also only 3 different light
intensities that were measured as the room light was used as a control.

Improvements:
The next time this experiment is conducted, more trials should be conducted in order to
reduce random and systematic errors and make the result more accurate with the
calculation of an average distance moved for each light intensity. More light intensities
could be used, varying the distance of the lamp. Different coloured lights (using filters)
could also be used as a variation to light intensity, to test the effect on the rate of
transpiration. In the dark room, the reading could be taken at room light, by removing the
photometer from the dark room so there is less light intensity as compared to the phone
light reducing the chances of transpiration occurring.