Testing a leaf for starch

Investigating photosynthesis – starch and chlorophyll

During photosynthesis a plant absorbs light energy using the pigment chlorophyll. This allows it to convert carbon dioxide and water into glucose. This glucose is:

  • transported to the growing parts of the plant for use in respiration
  • transformed into cellulose, proteins and oils
  • turned into starch for storage

Therefore, to test if a plant has been photosynthesising, you can test the leaf to see if starch is present.

Starch testing

Iodine solution is used to test leaves for the presence of starch. You need to:

  1. heat a plant leaf in boiling water for 30 seconds (this kills the leaf, stopping any chemical reactions)
  2. add the leaf to boiling ethanol in a water bath for a few minutes (the boiling ethanol dissolves the chlorophyll and removes the green colour from the leaf - it turns white so it is easy to see the change in colour)
  3. wash with water to rehydrate and soften the leaf and spread onto a white tile
  4. add iodine solution from a dropping pipette

After a few minutes, the parts of the leaf that contain starch turn the iodine from brown to blue/black.

Variegated leaves have green parts (where the cells contain chlorophyll) and white parts (where there is no chlorophyll). Only the parts that were green become blue/black with iodine solution, showing the importance of chlorophyll in photosynthesis. The parts without chlorophyll do not photosynthesise, and so they do not make starch and the iodine does not change colour.

Two variegated leaves. The left one has green parts which contain chlorophyll for photosynthesis. The right one has been treated with iodine. The parts containing chlorophyll turn blue-black.

The leaf on the left is a variegated leaf. The green parts contain chlorophyll and photosynthesise to make starch. The white part of the leaf does not contain chlorophyll, so does not photosynthesise.

The leaf on the right shows a positive test for starch in the areas which contained chlorophyll, and a negative test for starch in the areas which lack chlorophyll.

This is evidence that chlorophyll is required for photosynthesis.

A plant can be ‘de-starched’ by leaving it in the dark for a few hours. Parts of its leaves are covered with dark paper, and the plant is left in the light for a few hours. Only the uncovered parts become blue/black with iodine solution, showing the importance of light in photosynthesis.

Question

Ethanol is flammable and so is heated using a hot water bath instead of using a Bunsen burner. Because ethanol boils at 78°C, a tube of it boils when placed in a beaker of hot water.

In this experiment you need to place a leaf in boiling ethanol to decolourise it. Write a risk assessment for this.

HazardRiskControl measure
HazardRiskControl measure
Ethanol is flammable.It may ignite when heating to boiling point if using a naked flame.Do not heat over a Bunsen burner. Use a beaker of hot water as a water bath.