Exercise :- 4 Measurement of the rate of transpiration using

photometer

A potometer (from Greek ποτό = drunken, and μέτρο = measure) —

sometimes known as a transpirometer— is a device used for measuring the rate
of water uptake of a leafy shoot. The causes of water uptake are photosynthesis and
transpiration. Everything must be completely water tight so that no leakage of
water occurs.

There are two main types of potometers used - the bubble potometer (as
detailed below), and the mass potometer. The mass potometer consists of a plant
with its root submerged in a beaker. This beaker is then placed on a digital balance;
readings can be made to determine the amount of water lost by the plant. The mass
potometer measures the water lost through transpiration of the plant and not the
water taken up by the plant.
A potometer is a piece of apparatus used to measure the rate of water loss from a plant
(transpiration). The rate of transpiration can be estimated in two ways:
1) Indirectly - by measuring the distance the water level drops in the graduated tube over a
measured length of time. It is assumed that this is due to the cutting taking in water which in turn
is necessary to replace an equal volume of water lost by transpiration. 2) Directly - by measuring
the reduction in mass of the potometer over a period of time. Here it is assumed that any loss in
mass is due to transpiration.

Design
Potometer come in a variety of designs, but all follow the same basic principle.
•A length of capillary tube A bubble is introduced to the capillary; as water is taken up by the
plant, the bubble moves. By marking regular gradations on the tube, it is possible to measure
water uptake.
•A reservoir. Typically a funnel with a tap; turning the tap on the reservoir resets the bubble.
Some designs use syringe instead.
•A tube for holding the shoot. The shoot must be held in contact with the water; additionally,
the surface of the water should not be exposed to the air. Otherwise, evaporation will interfere
with measurements. A rubber bung greased with petroleum jelly suffices.
Preparation
Cut a leafy shoot from a plant and plunge its base into water. This prevents the xylem
from taking up any air. Wetting the leaves themselves will alter the rate of transpiration.
•Immerse the whole of the potometer into the sink. Move it about until all the air bubbles
come out.
•Recut the shoot's stem underwater. Put it into the bung; grease the bung with plenty of
petroleum jelly (Vaseline) if it doesn't stay and then put the bung into the potometer.
•Make sure the tap is closed, then lift the whole assembly out of the water.
•Leave the end of the capillary tube out of the water until an air bubble forms then put
the end into a beaker of water.
Types of Potometer
•Bubble Potometer
•Mass potometer
•Ganong's Potometer
•Darwin's Potometer
Use
•Set up the conditions of the experiment. Alterations to lighting (placing the plant in
bright light or shadow), wind (directing a fan at the plant), and humidity (placing the
plant in a humid chamber) are typical.
•Let the bubble reach a "zero" point in the tube.
•Measure the movement of the bubble at regular intervals and record the results
Precautions
•When a twig is cut from a plant, it should be immediately put under water (only the
cut portion). Then, a small part is cut under water. This prevents entry of air into the
xylem vessels.
•The conditions of the potometer, other than the alteration that is being tested, should
not be changed during a test, as outside conditions (for example, temperature)
determine water uptake.
Limitations
The Potometer does not measure the rate of transpiration accurately because not all
of the water that is taken by the plant is used for transpiration (water taken might be used
for photosynthesis or by the cells to maintain turgidity). The potometer measures the rate
of uptake of water. To measure transpiration rate directly, rather than the rate of water
uptake, utilize a scientific instrument which quantifies water transfer at the leaves.
The water retained by plant is so negligible that it can be neglected.
Introducing an air bubble may not be easy.
A twig may not be active for a long time.
A simple experiment is described below to demonstrate how the rate of transpiration
of common plants found in Hong Kong can be determined with the use of potometer.
Moreover, students can also be given the opportunity to estimate the stomatal density of
leaves from different plants to evaluate the relationship between different transpiration
rates and this morphological feature.
Estimation of transpiration rate using a potometer

Fig.1 Experimental setup

Procedure
1. Use a sharp razor blade to cut a leafy shoot under water.
2. Insert the leafy shoot through the hole of the stopper provided with the potometer.
3.Fill the potometer with water and fit the stopper holding the leafy shoot to the
apparatus.
4. Use vaseline to seal all the connections of the apparatus.
5. Trap an air bubble in the capillary tube by the following procedures:
• dip the end of the capillary tube into a beaker of water,
• close the tap of the reservoir,
• take away the beaker of water and allow the plant to transpire for a while, and
• re-immerse the capillary tube into the beaker of water again.
6. Wait for about 5 minutes for the plant to equilibrate.
7. Estimate the rate of transpiration by measuring the distance moved by the air bubble
per unit time. Take another measurement, and average the two readings.
8. Find out the total leaf surface area of the experimental plant using a graph paper.
9. Transpiration rate can be expressed in terms of water transpired per unit time per unit
area of leaf surface.
10. If time permits, estimate the rate under different environmental conditions, e.g. under
direct incandescent light illumination, under low light condition, in rapid air
movement (provided by a fan) or in still air.
Note
1. The total surface area of leaf should include the areas of both upper and lower surfaces.
2. This potometer only measures the rate of water uptake, it cannot find out the actual
transpiration rate.
3. The volume of water transpired can be estimated using the formula below:
Distance moved by air bubble = d cm
Cross-sectional area of capillary tube = a cm2
Volume of water transpired (V) = d x a cm3