INVESTIGATING RATE OF PHOTOSYNTHESIS

Starch Production & Need for Chlorophyll

 Starch is stored in chloroplasts where photosynthesis occurs so testing a leaf for

starch is a reliable indicator of which parts of the leaf are photosynthesizing.

 Leaves can be tested for starch using the following procedure:

 A leaf is dropped in boiling water to kill the cells and break down the cell
membranes.
 The leaf is left for 5-10 minutes in hot ethanol in a boiling tube. This removes the
chlorophyll so colour changes from iodine can be seen more clearly.
 The leaf is dipped in boiling water to soften it.
 The leaf is spread out on a white tile and covered with iodine solution.
 In a green leaf, the entire leaf will turn blue-black as photosynthesis is occuring in all
areas of the leaf.
 This method can also be used to test whether chlorophyll is needed for photosynthesis by
using a variegated leaf (one that is partially green and partially white).
 The white areas of the leaf contain no chlorophyll and when the leaf is tested only the
areas that contain chlorophyll stain blue-black.
 The areas that had no chlorophyll remain orange-brown as no photosynthesis is
occurring here and so no starch is stored.
 Care must be taken when carrying out this practical as ethanol is extremely
flammable, so at that stage of the experiment the Bunsen burner should be turned off.

 The safest way to heat the ethanol is in an electric water bath rather than using a beaker
over a Bunsen burner with an open flame.

The Need for Light in Photosynthesis

 The same procedure as above can be used to investigate if light is needed for
photosynthesis.
 Before starting the experiment the plant needs to be destarched by placing in a dark
cupboard for 24 hours.
 This ensures that any starch already present in the leaves will be used up and will not
affect the results of the experiment.
 Following destarching, a leaf of the plant can be partially covered with aluminium
foil and the plant placed in sunlight for a day.
 The leaf can then be removed and tested for starch using iodine.
 The area of the leaf that was covered with aluminium foil will remain orange-brown as
it did not receive any sunlight and could not photosynthesise, while the area exposed to
sunlight will turn blue-black.
  This proves that light is necessary for photosynthesis and the production of starch.

The Need for Carbon Dioxide in Photosynthesis

 Destarch a plant.
 Tie a clear bag containing sodium hydroxide, which will absorb carbon dioxide from
the surrounding air, around one leaf.
 Tie a clear bag containing water (control experiment), which will not absorb carbon
dioxide from the surrounding air, around another leaf.
 Place the plant in bright light for several hours.
 Test both leaves for starch using iodine.
 The leaf from the bag containing sodium hydroxide will remain orange-brown as it
could not photosynthesise due to lack of carbon dioxide.
 The leaf from the control bag containing water should turn blue-black as it had all
necessary requirements for photosynthesis.

Investigating the Rate of Photosynthesis

 The plants usually used are Elodea or Camboba - types of pondweed

 As photosynthesis occurs, oxygen gas produced is released 
 As the plant is in water, the oxygen released can be seen as bubbles leaving the cut end
of the pondweed
 The number of bubbles produced over a minute can be counted to record the rate
 The more bubbles produced per minute, the faster the rate of photosynthesis
 A more accurate version of this experiment is to collect the oxygen released in a test tube
inverted over the top of the pondweed over a longer period of time and then measure
the volume of oxygen collected

Investigating the effect of changing light intensity, by moving a lamp different distances
away from the beaker containing the pondweed:

 
 Investigating the effect of changing temperature on photosynthesis, by changing the
temperature of the water in the beaker

 
As temperature increases the rate of photosynthesis increases as the reaction is controlled by
enzymes. However, as the reaction is controlled by enzymes, this trend only continues up to a
certain temperature beyond which the enzymes begin to denature and the rate of
reaction decreases

Investigating the effect of changing carbon dioxide concentration, by dissolving different

amounts of sodium hydrogen carbonate in the water in the beaker 
 

Investigating the effect of changing carbon dioxide concentration on the rate of

photosynthesis

 Care must be taken when investigating a condition to keep all other variables
constant in order to ensure a fair test
 For example, when investigating changing light intensity, a glass tank should be placed
in between the lamp and the beaker to absorb heat from the lamp and so avoid changing
the temperature of the water as well as the light intensity


 INVESTIGATING NET GAS EXCHANGE

 During the day, especially when the sun is bright, plants are photosynthesising at a
faster rate than they are respiring, so there is a net intake of carbon dioxide and a
net output of oxygen
 We can investigate the effect of light on the net gas exchange in an aquatic plant using
a pH indicator such as hydrogencarbonate indicator 
 This is possible because carbon dioxide is an acidic gas when dissolved in water
 Hydrogencarbonate indicator shows the carbon dioxide concentration in solution
 The colour of the indicator turns yellow or orange at higher levels of carbon dioxide
concentration (respiration > photosynthesis means lower pH).
 The colour of the indicator turns purple at low levels of carbon dioxide concentration
(photosynthesis > respiration means higher pH)

 Several leaves from the same plant are placed in stoppered boiling tubes containing
some hydrogencarbonate indicator 
 The effect of light can then be investigated over a period of a few hours
 Results from a typical experiment are shown in the table below:

TUBE CONTENTS CONDITIONS INDICATOR CONCLUSION

TURNS
A Leaf Light Purple There is a net intake of oxygen by a
leaf in light.
B Leaf Dark Yellow There is a net intake of carbon-
dioxide by a leaf in the dark
C No leaf Light Red This is the control.The two other
tubes can be compared with it.
 INVESTIGATING RATE OF TRANSPIRATION

Investigating the role of environmental factors in determining the rate of transpiration

from a leafy shoot

 Cut a shoot underwater to prevent air entering the xylem and place in tube
 Set up the apparatus as shown in the diagram and make sure it is airtight, using vaseline
to seal any gaps
 Dry the leaves of the shoot (wet leaves will affect the results)
 Remove the capillary tube from the beaker of water to allow a single air bubble to form
and place the tube back into the water
 Set up the environmental factor you are investigating
 Allow the plant to adapt to the new environment for 5 minutes
 Record the starting location of the air bubble
 Leave for a set period of time
 Record the end location of air bubble
 Change the light intensity or wind speed or level of humidity or temperature (only one -
whichever factor is being investigated)
 Reset the bubble by opening the tap below the reservoir
 Repeat the experiment
 The further the bubble travels in the same time period, the faster transpiration is
occurring and vice versa
 The further the bubble travels in the same time period, the faster transpiration is
occurring and vice versa

Environmental factors affecting rate of transpiration can be investigated in the following

ways:

 Temperature : Temperature of room (cold room and warm room). Rate of transpiration
increases with increase in temperature.
 Humidity : Spray water in plastic bag and wrap around plant. Rate of transpiration
decreases with increase in humidity

 
Investigating transpiration rates using a potometer