PHOTOSYNTHESIS

Objectives:
After working on this exercise, you should be able to:

1. Explain how different factors affect photosynthesis

2. Discuss the basis of paper chromatography
3. Relate the properties of light to the existence of different pigments in plants.
4. Explain the significance of photosynthesis in the biosphere.
5. Describe the overall process of photosynthesis.
Materials:

San Francisco (Coleus leaves) tap water

Hydrilla sp. (Digman) chromatography paper
5 mature papaya leaves test tubes
Cork cooled boiled water
Drinking straw mortar and pestle
2-week-old potted bean plant solvent (95 parts petroleum ether and
5 parts acetone)
Petri dish iodine solution
Carbon paper 95% ethyl alcohol
Hot plate phenol red
Beaker paper clip or adhesive
Forceps pencil
Paper towels

Procedures

A. Effect of Chlorophyll
1. Select one young variegated leaf of Coleus. Examine and take note of the
areas with chlorophyll. Sketch and label the green and non – non green areas.
2. Boil the leaf 2 – 3 minutes.
3. Remove the leaf, blot it dry and immerse it in 95% ethyl alcohol until the
pigments are extracted.
4. Rinse the bleached leaf with tap water. Place in a petri dish and 2-3 drops of
dilute iodine solution to determine the presence of starch. The occurrence of
photosynthesis is indicated by a bluish – black to purple coloration.
5. Sketch the bleached leaf and label the photosynthetic and non –
photosynthetic portions.
B. Effect of Light
1. Grow two potted bean plants for 1-2 weeks. Place them in the dark for 48-
72 hours.
2. Cover one plant with carbon paper and keep the second plant
uncovered. Expose both plants to light for 6 hours.
3. Detach one leaf each from the covered and uncovered plants. Test for the
presence of starch by adding three drops of iodine solution per leaf.

C. Effect of Carbon Dioxide

1. Obtain three test tubes and number them from 1 to 3.
2. Fill half of each tube with cooled boiled water.
3. Add five drops of phenol red to each tube and shake gently.
4. With a drinking straw, blow into the water in test tubes 1 and 2 until the
solutions show a change in color from green to yellow.
5. Place a shoot of Hydrilla in test tube 1. Place the three test tubes in very
bright light. After 20 – 30 min, observe the water for color changes.

D. Separation of Chloroplast Pigments by Paper Chromatography

1. Obtain five mature leaves of Carica papaya.
2. Cut the leaves into small pieces and grind them to obtain a deep green
liquid extract. This is your chlorophyll extract.
3. Obtain a strip of chromatography paper big enough to fit into a test tube
that will serve as a chromatography chamber.
4. Using a pencil, mark a spot about 1 cm from the base of the paper.
5. Place 3 – 5 drops of chlorophyll extract on the spot, making sure that the
spot dries quickly between applications.
6. Carefully hang the strip on the lower or basal end of a cork with the use of
a paper clip or adhesive, and lower this into a test tube containing 3 -5 ml
of solvent (95 parts of petroleum ether and 5 parts of acetone). The tip of
the paper should be in contact with the solvent
7. Observe for the movement of solvent up the strip. Make frequent
observations, or else some pigments may become superimposed on each
other if the separation continues for too long.
8. Stop the development of the chromatogram when the solvent has traveled
to above 1 -2 cm form the top of the strip.
9. Sketch the outline of the chromatogram.
10. Determine the RF value for each pigment.

(Retention factor) RF= DISTANCE OF SOLUTE/DISTANCE MOVED BY PIGMENT

DISTANCE OF SOLVENT
Data and Observations:

A. Effect of Chlorophyll

A.1. Variegated leaf (not bleached) A.2. Bleached lead

Description: Description:

B. Effect of Light
Reaction to starch test: leaf from covered plant: ____________
Leaf from uncovered plant: _________
C. Effect of Carbon Dioxide
Color changes in test tubes with Phenol red
Test tube Before blowing After blowing 20 -30 minutes
after adding
Hydrilla
1
2
3

D. Separation of Pigments by Paper Chromatography

*outline of chromatogram

RF Values:
Pigment 1 _________________
Pigment 2_________________
Pigment 3_________________
Pigment 4 _________________