Practical 6: To investigate conditions essential for photosynthesis Objective: (a) To test the presence of starch in green leaf (b)

To show that sunlight is necessary for photosynthesis (c) To show that chlorophyll is essential for photosynthesis (d) To show that carbon dioxide is necessary for photosynthesis (e) To show that oxygen is produced during photosynthesis. Material and apparatus: Leaves Hydrilla or any water plant Beaker Bell jar Filter funnel Polythene bag Test tube White tile Methanol Iodine solution Boiling water Calcium hydroxide Natrium hydroxide

Procedure: (a) To test the presence of starch in green leaf 1. A leaf was plucked from a plant that has been exposed to sunlight for a few hours. 2. The leaf was placed in boiling water. This is to kill and to soften the leaf. 3. A small amount of methanol was placed in a boiling tube. The leaf was placed into it. 4. The tube was placed in the beaker of hot water. 5. The leaf was pulled out after 5 minutes to check the leaf was decolourised. 6. The leaf was dipped into hot water for a few seconds. 7. The leaf was lay on a white tile and three or four drops of iodine was putted on it 8. The observation was recorded in the table below.

Results: Contents Leaves Observation A blue-black colourisation Inference Starch produced when expose to sunshine in the process of photosynthesis

(b) To show that sunlight is necessary for photosynthesis 1. A potted plant in the darkness for about 24 hours. 2. A leaf was plucked from the plant.

3. The leaf was tested for starch by repeating the step 2-6 in (a) 4. The observation was recorded in the table

Results: Content Leaves Observation The colour of iodine remains yellow. Inference Starch is not produce without the present of sunshine.

(c)

To show that chlorophyll is essential for photosynthesis 1. A variegated leaf was removed from a plant that has been exposed to sunlight for a few hours. 2. A diagram was drawn to show the distribution of the green colour in the leaf. 3. The leaf was tested for starch. Step 2-6 was repeated as in (a) 4. The observation was recorded in the table below.

Results: Content Leaves Observation Green colour of the leave will give blue-black colourisation with iodine test but other colour of the leave will remain with the yellowish colour from the iodine Inference Starch only present in green leaves which have chlorophyll. .

(d) To show that carbon dioxide is necessary for photosynthesis. 1. Two potted plants were left in the darkness for about one day to remove any starch present. 2. The pots were covered with polythene bags. 3. The plants are places under two bell jars, which labelled A and B respectively. 4. A beaker of potassium hydroxide was putted in bell jar A. 5. An open tube was used in bell jar B. 6. The two bell jars were placed under sunlight for a few hours. 7. The leaf was tested for starch. Step 2-6 was repeated in (a) 8. The observation was recorded in the table. Results: Jar A Observtion The colour of iodine remains yellow Inference Strarch does dot produce without the presence of carbon dioxide. B Blue-black colouration Starch produce with the present of carbon dioxide.

(E) To show that oxygen is produced during photosynthesis. 1. Some freshwater plants (Hydrilla) were placed under an inverted filter funnel in a beaker of water, preferably pond water. The funnel was raised above the bottom of the beaker to allow free circulation of water. 2. A test tube completely filled with water over the stem of the funnel was inverted. The set was labelled as A

3. The apparatus was placed under the sunlight for a few hours. 4. Repeat the procedure 1- with other plants. The plants were placed in a dark cupboard. The set was labelled as B. 5. The gas in the test tube of Set A and Set B was tested with glowing splint. 6. The observation was recorded in the table below. Results: Set A Observation Glowing splint will be fire Inference Oxygen is contained in the test tube Oxygen is produced during photosynthesis. B No reaction on glowing splint Oxygen is not contained in the test tube Oxygen cannot be produced without sunshine.

Questions: 1. What do you observe concerning the distribution of the blue black colour in the leaf? Green colour of the leaf will give blue black colourisation when doing the iodine test but the other colour of the leave will remain tallow colour of iodine. 2. Can you say that only the green portions of the leaf manufacture starch in the sunlight? Yes, photosynthesis only occurs when chlorophyll is present with the present of sunlight, carbon dioxide (Co2), suitable temperature and water.

Discussion: Photosynthesis is the process of converting light energy to chemical energy and storing it in the bonds of sugar. This process occurs in plants and some algae (Kingdom Protista). Plants need only light energy, CO2, and H2O to make sugar. The process of photosynthesis takes place in the chloroplasts, specifically using chlorophyll, the green pigment involved in photosynthesis. 6H2O + 6CO2 ----------> C6H12O6+ 6O2 Photosynthesis takes place primarily in plant leaves, and little to none occurs in stems, etc. The parts of a typical leaf include the upper and lower epidermis, the mesophyll, the vascular bundle(s) (veins), and the stomates. The upper and lower epidermal cells do not have chloroplasts, thus photosynthesis does not occur there. They serve primarily as protection for the rest of the leaf. The stomates are holes which occur primarily in the lower epidermis and are for air exchange: they let CO 2 in and O2 out. The vascular bundles or veins in a leaf are part of the plant's transportation system, moving water and nutrients around the plant as needed. The mesophyll cells have chloroplasts and this is where photosynthesis occurs. The main factors are light intensity, carbon dioxide concentration and temperature, known as limiting factors. As light intensity increases, the rate of the light-dependent reaction, and therefore photosynthesis generally, increases proportionately. As light intensity is increased however, the rate of photosynthesis is eventually limited by some other factor. Chlorophyll a is used in both photosystems. The wavelength of light is also important. PSI absorbs energy most efficiently at 700 nm and PSII at 680 nm. Light with a high proportion of energy concentrated in these wavelengths will produce a high rate of photosynthesis. An increase in the carbon dioxide concentration increases the rate at which carbon is incorporated into carbohydrate in the light-independent reaction and so the rate of photosynthesis generally increases until limited by another factor.

Photosynthesis is dependent on temperature. It is a reaction catalysed by enzymes. As the enzymes approach their optimum temperatures the overall rate increases. Above the optimum temperature the rate begins to decrease until it stops. Conclusion: (a) Starch will be produced in the present of carbon dioxide (b) The leaf that was not exposed to sunshine cannot produce starch because it cannot undergo photosynthesis. (c) The plant which has green pigment will undergo photosynthesis to produce starch. (d) Starch will produce in the present of carbon dioxide. (e) Exposure of the green water plant to sunlight caused oxygen to be produced.