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Investigating the need for chlorophyll, light and carbon dioxide

Plant nutrition · Photosynthesis · note 4 of 7

Investigating the need for chlorophyll, light and carbon dioxideSpec 6.1.7

In short

Testing a leaf for starch shows whether it has photosynthesised, because glucose made in photosynthesis is converted to starch. The plant is destarched in the dark for 24 to 48 hours first. The leaf is boiled, then heated in ethanol to remove the chlorophyll, and iodine turns from orange-brown to blue-black where starch is present.

To show that chlorophyll, light and carbon dioxide are needed for photosynthesis, test leaves for starch. Glucose made in photosynthesis is converted to starch, so starch in a leaf is evidence that photosynthesis has taken place.

Destarching

Before each investigation, destarch the plant by leaving it in the dark for 24 to 48 hours. The plant uses up its stores of starch in respiration. Leaves from a destarched plant test negative for starch.

The starch test on a leaf

  1. Drop the leaf into boiling water for about 30 seconds. This kills the cells and softens the leaf.
  2. Turn off the Bunsen burner. Put the leaf in a boiling tube of ethanol and stand the tube in a beaker of hot water for several minutes. The ethanol removes the chlorophyll, so the leaf turns pale.
  3. Dip the leaf back into the hot water to soften it, then spread it out on a white tile.
  4. Add a few drops of iodine solution. Starch turns the iodine from orange-brown to blue-black.

Investigating each requirement

Investigating what photosynthesis needs
To investigateMethodControlResult
ChlorophyllUse a variegated leaf (green and white patches). Destarch, expose to light, then test for starch.The green parts of the same leafOnly the green parts, which contain chlorophyll, turn blue-black. The white parts stay orange-brown.
LightCover part of a destarched leaf with foil or black paper, leave the plant in the light, then test for starch.The uncovered part of the same leafThe uncovered part turns blue-black. The covered part stays orange-brown.
Carbon dioxidePut a destarched plant in a sealed bag or bell jar with soda lime (or potassium hydroxide), which absorbs carbon dioxide, and leave in the light. Test a leaf for starch.An identical plant sealed in a bag with no soda limeThe leaf without carbon dioxide stays orange-brown. The control leaf turns blue-black.
Practical skill:

Use a control so that only one factor changes. In the variegated leaf and foil investigations the control is part of the same leaf, so everything else is identical. Ethanol is flammable: switch off the Bunsen burner before heating it, use a hot water bath, and wear eye protection.

Exam tip:

If a question asks why the leaf was heated in ethanol, the answer is to remove (decolourise) the chlorophyll so the colour change with iodine can be seen.

Testing a leaf for starch in three steps: the leaf in boiling water over a Bunsen burner, the leaf in a boiling tube of ethanol standing in a beaker of hot water with the Bunsen burner turned off, and iodine solution dropped onto the leaf on a white tile. Below are the results: a variegated leaf turns blue-black only where it was green, a leaf partly covered by foil turns blue-black only where it was uncovered, and a plant sealed in a bag with soda lime gives an orange-brown leaf while the control plant gives a blue-black leaf. (opens full size in a new tab)
The starch test, and the results that show photosynthesis needs chlorophyll, light and carbon dioxide.

Written and checked against the Cambridge IGCSE Biology (0610) specification · Updated October 2026

Frequently asked questions

What are the products of photosynthesis used for?

The glucose made in photosynthesis is used for respiration, stored as starch, built into cellulose for cell walls, converted to sucrose for transport in the phloem, and used to make nectar to attract insects. Oxygen is released as a by-product. Glucose is also combined with nitrogen from nitrate ions to make amino acids.

How does light intensity affect the rate of photosynthesis?

Supplement As light intensity increases, the rate of photosynthesis increases, because light provides the energy for the process. The rate then levels off, because another factor, such as carbon dioxide concentration or temperature, becomes limiting. You can show this by moving a lamp closer to pondweed and counting the oxygen bubbles released per minute.

How does temperature affect the rate of photosynthesis?

Supplement As temperature rises, the rate of photosynthesis increases up to an optimum, because photosynthesis is controlled by enzymes and molecules move faster with more collisions. Above the optimum, the rate falls sharply because the enzymes are denatured. On a cold bright day, temperature is likely to be the limiting factor.

All 5 questions on Photosynthesis