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Effect of Light Intensity

on the Rate of Photosynthesis

Light intensity is one of the factors affecting the rate of photosynthesis. Other factors are
concentration of carbon dioxide, temperature and to a lesser degree, water. Light intensity
directly affects the light-dependentreaction in photosynthesis and indirectly affects the light-
independent reaction.

Light is a limiting factor when the light intensity is too low to allow the light-dependent
reaction to proceed at its maximum rate. Light is not normally a limiting factor except for plants
in forest understory of other shaded plants. Very high light intensities may slow the rate of
photosynthesis due to bleaching of chlorophyll. However, plants exposed to such conditions
usually have protective features to include thick, waxy cuticles and hairy leaves.

Measuring the rate of oxygen evolution using a water plant is commonly used to measure the rate
of photosynthesis. Oxygen is a gaseous product, so can be measured by noting volume changes
of the number of bubbles evolved (less accurate).

Apparatus and Materials

Mercury/projector lamp e.g. 60 W


Stop watch

Apparatus for collecting gas e.g. graduated syringe connected via tubing (This is
optional. In some practical exercise the rate of oxygen production is determined -
counting number of bubbles per minute)

400 cm3 beaker as water bath/heat filter


Test tube containing dilute sodium hydrogencarbonate solution

Pondweed eg Elodea, Hydrilla

1. Cut the stem of a bubbling pond weed which has been well illuminated to about 5cm. Place
the cut surface upwards in a test tube containing sodium hydrogencarbonate solution.

2. Place the test tube in the beaker of water and note the temperature. The beaker of water acts
as a heat filter or heat shield, so its temperature should be checked at intervals to ensure that it is
constant throughout the experiment; the water should be renewed if necessary.

3. Attach the gas-collecting apparatus, if any.

4. Darken the laboratory by turning off as many lights as possible.

5. Place lamp 10 cm away from the beaker. Allow the plant to equilibrate or adjust to the light
intensity for 2-3 minutes
When the rate of air bubbles is regular and a adequate (>10 bubbles/minute), place the capillary
tube/test tube over the cut tip of the pondweed and then measure the volume. OR, count the
number of bubbles. This should be done for 5 minutes. Repeat twice and obtain an average of
the results.

6. Repeat steps 4 and 5, with increasing distances away from the light source e.g. 20 cm, 30 cm,
40 cm and 50 cm. Light intensity is a inversely proportional to the square of the distance, so as
the distance is increased the light intensity decreases. Note that doubling the the distance does
not half the intensity, rather quarters it.

7. Record results in a table, then plot a graph of volume of oxygen/minute OR number of

bubble/minute against the distance between the lamp and the plant.

As light intensity increases (distance between lamp and plant decreases) the volume of oxygen
(or the rate of bubble production) increases. This indicates that the rate of photosynthesis
increases with light intensity.

However, at sufficiently high levels of light intensity, the rate oxygen evolution remains constant.

Use the Waterweed Simulator to manipulate the light intensity, wavelength and carbon dioxide
and see their effect on the rate of photosynthesis.

1. Why is sodium hydrogen carbonate solution used instead of water?

Sodium hydrogencarbonate increases carbon dioxide availability, and ensures that carbon
dioxide is not a limiting factor for the the reaction. Plain water water may not contain enough
oxygen so that the maximum rate of photosynthesis can occur.

Ideally, pondwater should be used to make up the sodium hydrogencarbonate solution. Tap
water may be used to make the solution, but only after it has been left to stand overnight, so that
it chlorine content is reduced. Chlorine tends to inhibit the rate of photosynthesis.

2. In some cases, a trace of detergent is also added to the water in the test tube. Explain
Detergent may be added to reduce surface tension. This will allow bubbles to escape
freely. When surface tension is high, bubbles are of varied sizes.

3. Suggest how this procedure can be modified

that the distance between the lamp and the p
not varied.
The power of the lamp may be varied e.g. use of
40W, 60W and 100W bulbs.

4. Why was the laboratory darkened?

The laboratory was darkened to reduce light from
sources, toensure that the light intensity was ma
to the lamp.

5. What are the main sources of inaccuracy in the experiment?

When counting bubbles, the bubbles may be of varied sizes, leading to inaccurate conclusions
about the rate of oxygen production. If the rate of bubbling is very high, then there may be errors
in counting. Another source of inaccuracy is the presence of other light sources, of which
intensities may be varied by shifting shadows.

5. How can we prove that oxygen was the gas produced?

A glowing splint should be placed near the mouth of the test tube, where the gas was collected.If
oxygen is present the splint is re-ignited into a flame.

6. If the gas which was produced was analyzed and found not to be pure oxygen, how
would you account for this?
Water contains other dissolved gases which came out of solution and mixed with the
oxygen. Carbon dioxide is expected in this mixture, particularly if sodium hydrogen carbonate
was used.

7. What is the relationship between the light intensity and the rate of photosynthesis at low
light intensities?
At low light intensities, the rate of photosynthesis increases with the intensity; the rate is directly
proportional to intensity ( but inversely proportional to distance)

8. Why does the rate of photosynthesis not increase at very high light intensities?
At very high light intensities, the rate of photosynthesis levels off or remains constant. At this
point, another factor - other than light - contributing to the rate of photosynthesis is limiting. The
most likely reason for the leveling off of the reaction rate is the saturation of active sites in the
enzymes catalyzing the reactions.

In the case of this experiment - the limiting factors are unlikely to be water or carbon dioxide. A
limiting factor may be temperature - a slight increase in the ambient temperature may result an
increase in the rate of reaction. However, a high temperatures the enzymes catalyzing the
photosynthetic reactions would be denatured.