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Exercise 13A The Role of Chlorophyll in Photosynthesis Group 1: Abid, Latoga, Pidlaoan, Rodriguez Bio 10 Lab 1A, Sir

Jeremy Vicencio I. Abstract Photosynthesis is one of the most important processes on Earth. It Photosynthesis is a process wherein plants absorb light energy through the chlorophyll, found in leaves, to convert carbon dioxide (CO2) and water (H2O) to glucose (C6H12O6), their form of food, and release oxygen gas (O2) as a by-product. If the plant produces extra glucose, it is stored in its tissues in the form of starch. Therefore, testing the leaf for the presence of starch rather than testing it for glucose is an effective way of knowing whether it has been performing photosynthesis or not. This may be done by staining the leaf with Lugols solution (I2KI), thus forming a deep blue color when the amylase in starch gets in contact with iodine. II. Introduction Photosynthesis is the conversion of light energy to chemical energy, which is stored in sugar and other organic molecules. Photosynthesis may be oxygenic or anoxygenic. In oxygenic photosynthesis, carbon dioxide (CO2) and water (H2O) are used to make glucose, with oxygen gas (O2) as the by-product. This process occurs in higher plants (e.g., rice, maize, mosses, ferns, forest trees, shrubs, etc), in some algae (brown, green, red, yellow) and in blue-green cyanobacteria. In anoxygenic photosynthesis, a variety of reducing chemicals (such as H2S, H2, Fe2+, S0, organic carbon) are used instead of water, thus generating sulfur, sulfate, or iron as the by-product. This occurs in other kinds of bacteria (green and purple), heliobacteria, and some cyanobacteria. The process of photosynthesis in plants takes place in the chloroplasts, which are found in the interior tissue of the leaf called the mesophyll, using the green pigment chlorophyll. Photosynthesis occurs in two steps. First, the light reactions, which take place in the thylakoid membranes, split water, thus releasing O2, and producing ATP and NADPH. The Calvin cycle, which is carried out in the stroma of the chloroplast, form sugar from CO2 using the ATP and NADPH produced in the light reactions. Photosynthesis in plants may be summarized as: 6 CO2 + 12 H2O + Light energy ---> C6H12O6 + 6 O2 + 6 H2O When there is excess glucose produced, it is converted to starch, which is temporarily stored in the chloroplasts. Chlorophyll is a green pigment found in the chloroplasts of green plants. It is a photoreceptor with a structure of a porphyrin ring with magnesium as its central ion. There are two main types of chlorophyll, chlorophyll a and chlorophyll b. These two types differ only in the composition of a side chain (CH3 in a, CHO in b). This structural difference is enough to cause the slight difference in the wavelengths of light absorbed in the red and
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Comment [JV1]: Total Score: 90/100

Comment [JV2]: 12/15

Comment [JV3]: Introduction a tad lengthy. Focus on specifying the objectives and hypotheses of the experiment and include only the necessary background information. 7/10

blue spectrum. This results to chlorophyll a being blue green in color while chlorophyll b is olive green. Chlorophyll converts the light energy it absorbs to chemical energy during photosynthesis. III. Methodology A sarasa (Graptophyllum pictum) leaf was acquired by the group. The leaf was boiled in water for ten minutes. The boiled leaf was then immersed in a test tube halffilled with 95% ethanol. A hot water bath was prepared, in which the test tube containing the boiled leaf immersed in ethanol was placed until all the pigments were extracted. The bleached leaf was then washed with water and tested for the presence of starch using Lugols solution (I2KI solution). IV. Results

parts of the leaf turned blue-black after staining with Lugols solution. Furthermore, there was no reaction in the parts of the leaf that used to be white. V. Discussion In the experiment, a variegated leaf, a leaf that has a variety of color caused by the difference in the amount of chlorophyll in some parts, was used to see the role of chlorophyll in photosynthesis. Since the main product of photosynthesis is glucose, the presence of starch, a carbohydrate composed of a large number of glucose, would indicate that photosynthesis took place. The leaf was boiled first in order to kill the leaf. This would stop the chemical reactions happening inside the leaf, break apart the cell membranes and soften the cell wall and cuticle. This makes it possible for hot ethanol to extract the chlorophyll and allow the penetration of the Lugols solution to the cells. The leaf was then immersed in ethanol to break down the chlorophyll, thus bleaching it. This was done to show the effect of Lugols solution more clearly. Also, since ethanol is an organic solvent, it properly extracts the chlorophylls form the leaf. The bleached leaf was then rinsed with water in order to remove the excess ethanol. This would also ensure that the Lugols solution would properly penetrate the leaf and would yield accurate results. The leaf was then stained with Lugols solution. Parts that turned blue-black in color indicate the presence of starch because the amylase in starch forms a deep blue color in the presence of iodine. The results of the experiment show that only the green parts of the leaf turned
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Comment [JV6]: 38/40 Comment [JV4]: 10/10

Comment [JV5]: 20/20

Comment [JV7]: Explain how starch reacts with Lugols solution to produce the characteristic blueblack color.

Figure 1: A sarasa leaf before boiling (above) and after soaking in Lugols solution (below)

Figure 1 shows the sarasa leaf before and after the treatment. The original green

blue-black after treating with Lugols solution. This means that the white parts of the leaf did not contain chlorophyll because there is an absence of starch, which then applies that photosynthesis does not take place in this certain part of the leaf. VI. Conclusion and Recommendations Since starch is made only in the parts of the leaf that has chlorophyll, it implies that chlorophyll is important and necessary for photosynthesis. The test for starch in the leaf is very effective and the experiment was a success. To further improve this experiment, the researchers recommend testing for other variegated plants in order to yield more results and observe more the importance of chlorophyll in photosynthesis. Examples of other variegated plants are Calathea makoyana, Lamium galebdolon and Cornus alba. The students should also exercise caution especially while doing the hot water bath. The ethanol may tend to boil unevenly and cause superheating. This can be dangerous for the ethanol solution may boil violently. This can be prevented by adding boiling chips to the ethanol solution. VII. References Biology Study Guide (2013). Testing the leaf for the presence of starch. Retrieved from http://brilliantbiologystudent.weebly. com/testing-a-leaf-for-the-presenceof-starch.html Carter, J. (2004). Photosynthesis. Retrieved from 104/photosyn.htm

Chemical of the Week (2013). Chlorophyll. Retrieved from ek/chlrphyl/chlrphyl.html Department of Biology. Biology 10 (General Biology) Laboratory Manual Govindjee & Whitmarsh, J. (1995). The Photosynthetic Process. Retrieved from e/paper/gov.html Home Science Tools (2013). Photosynthesis: Testing for starch. Retrieved from arch-test/a/1497/ Ingram, N., & Roberts, M. (2001). Nelson science: Biology (2nd ed.). UK: Nelson Thornes Ltd. May, P. (2013). Chlorophyll. Retrieved from orophyll/chlorophyll_h.htm Monger, G., Reiss, M., & Roberts, M. (2000). Advanced biology. UK: Nelson 2000. Reece, J., Urry, L., Cain, M., Wasserman, S., Minorsky, P., Jackson, R. (2009). Biology (9th ed.). CA: Pearson. Society of Biology (2011, November 24). Testing leaves for starch: The technique. Retrieved from actical-biology/testing-leaves-starchtechnique Steer, J. (2013). Structure and Reactions of Chlorophyll. Retrieved from /steer/chloro.htm

Comment [JV8]: 3/5 Good amount of references but observe parenthetical citation in the future.

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