Practical Investigation: Testing for starch in plants.

Photosynthesis is the process in which green plants (primarily) convert energy

from the sun’s light into usable, chemical energy.

Plants require energy for growth, reproduction, and defense. Excess energy,
created from photosynthesis, is stored in plant tissue as starch.

Starch is a white and powdery substance. It houses glucose, which plants use for
food.

The presence of starch in a leaf is reliable evidence of photosynthesis. That

is because starch formation requires photosynthesis.

MATERIALS AND APPARATUS:

• gloves
• a range of pot plants that can be easily moved around
• 100 ml beaker or glass jar in a saucepan with water
• bunsen burner, or a stove
• tweezers
• ethyl alcohol (or methylated spirits)
• glass petri dishes, white saucer or white tile
• stopwatch or timer
• glass pipette or dropper
• iodine solution

METHOD:
1. Work in groups of four or five.
2. Place half of the plants in the dark for at least 24-48 hours and the others
in a well-lit area of the class that is exposed to lots of natural sunlight.

3. After 24 hours, pour 50 ml of the methylated spirit into the beaker and
place it in the saucepan with water. Heat the saucepan over the bunsen
burner or the stove. The water in the saucepan will distribute the heat
evenly to warm the methylated spirit evenly.
4. Remove one healthy looking leaf from the pot plants that were in
the well-lit area exposed to direct sunlight.
5. Using the tweezers, dip a leaf into the boiling water for 1-2 minutes.
This helps to remove the waxy cuticle that covers the leaf and breaks
down the cell walls.
6. After this, place the leaf into the beaker with the methylated spirit.
7. Leave the leaf in the methylated spirit until all the chlorophyll has been
removed from the leaf and the methylated spirit turns green.
8. Place the leaf into warm water to soften it.
9. Remove the leaf from the warm water and place it on a white tile or a
petri dish on top of a white surface.
10. Use the pipette or dropper to carefully drop 2 or 3 drops of iodine
solution on the leaf in the petri dish and record your observations.
11. Repeat this process for two more leaves that were in the well-lit area.

Why is starch tested for and not glucose?

Glucose is the product of photosynthesis, and is rapidly converted to granules of

starch - a polymer of glucose - for storage. Starch granules have been visualized in
the stroma of the chloroplast as well as the cytoplasm.

It is advantageous to the plant to convert glucose to starch. Starch is insoluble in water

and therefore not a solute in the solution component of the stroma. Glucose on the
other hand is soluble in water and if it accumulated in the stroma, the stroma would
become more concentrated - hypertonic - relative to the cytoplasm. Water would move
from the cytoplasm ( higher water potential) via osmosis into the stroma of the
chloroplasts (lower water potential), causing them to swell and burst.

What observations will you make if the test is positive?

The iodine solution will turn from brown to blue - black.

Explain the significance of boiling the leaf in water.
Boiling the leaf in water:

• Removes the waxy cuticle which prevents entry of iodine/potassium iodide

solution.
• Ruptures cell membranes to make starch granules in cytoplasm and chloroplasts
accessible to iodine/potassium iodide solution. Cell membranes are selectively
permeable and do not readily allow the penetration of iodine.
• Denatures enzymes, particularly those which convert starch to glucose e.g.
diastase. Boiling arrests all chemical reactions, since enzymes which catalyse
the reactions are denatured. Denatured enzymes have altered or destroyed
active sites due to heat, pH, ionic concentration

Account for the texture of the leaf after being boiled in water.

The leaf was flaccid (soft) as cell membranes were ruptured and turgor pressure was
lost.