A Foray into Photosynthesis Austin Cao, AP Biology Mr.

Savage 11/1/13

Introduction Photosynthesis is the process by which organisms convert light energy into usable chemical energy that maintains life. First, we created a paper chromatography strip using spinach to reveal the various pigments used in photosynthesis and the distances they travelled. Second, we measured light transmittance (spectrophotometer) to compare the rates of photosynthesis of various solutions of boiled/unboiled chloroplasts under light/dark conditions. Third, we looked at other factors of interest that may affect photosynthesis by the floating leaf disk technique, since as oxygen is produced as a product, the leaves will float as the process occurs.

Chromatography

Band 1 2 3 4 5

Distance (mm) 7 19 29 41 90

Color olive green bright green yellow yellow orange white?

Identity cb ca x bc

Figure 1. Bands on chromatography strip made with spinach.

1) What factors are involved in the separation of the pigments? The only factors that determine separation of pigments are all properties of pigment molecules. For example: the molecules solubility, the molecules interaction with paper, and the size of the molecule.

2) Would you expect the Rf value to be different with a different solvent? Yes, this will affect how the molecules polarity interacts with the solvent when travelling through the paper.

3) Why do the pigments become separated during the development of the chromatogram? The pigment molecules travel with the water because they are water soluble. The reason they separate into bands is because the size of the pigment molecule determines how high it can climb through the paper.

Spectrophotometry time (minutes) Unboiled/dark Unboiled/light Boiled/light No chloros/light 0 21.80 40.45 7.95 12.25 5 40.32 88.45 9.35 12.13 10 (missed) 15 25.85 89.71 9.29 12.03 20 88.7 91.2 8.45 12.04

Figure 2. Lab data for transmittance percentages over time.

Transmittance over time for various environments

100 Transmittance (%) 80 60 40 20 0 0 5 15 20 Time (minutes) Unboiled/dark Unboiled/light Boiled/light No chloros/light

Figure 3. Lab data graphed.

1) What is the function of DPIP in this experiment? DPIP takes the place of NADP as the electron acceptor in photosynthesis. As the DPIP is reduced, it changes colors from blue to clear. Thus, DPIP allows us to practical method of observing the rate of photosynthesis. 2) What molecule found in chloroplasts does DPIP replace in this experiment? NADP is replaced.

3) What is the sources of the electrons that will reduce DPIP? Through hydrolysis, electrons are taken from water.

4) What was measured with the spectrophotometer in this experiment? The spec measures light transmittance, which we know is a measure of the extent of photosynthesis.

5) What is the effect of darkness on the reduction of DPIP? The lack of light should prevent photosynthesis and the reduction of DPIP from occurring.

6) What is the effect of boiling the chloroplasts on the subsequent reduction of DPIP? Boiling the chloroplasts should have denatured the molecules enough so that photosynthesis cannot occur.

7) What reasons can you give for the difference in the percentage of transmittance between the live chloroplasts that were incubated in the light and those that were kept in the dark? Light energy is required to excite electrons, thus allowing for the DPIP to be reduced. The reaction should have proceeded normally when incubated in light.

Floating leaf disc assay Abstract Rate of photosynthesis can be measured by through the floating disc method. We sought to measure the effect of different light frequencies on photosynthesis by incubating leaf discs in white light, red light, and no light. Both red and no light stopped the photosynthesis reaction, while white light resulted in a rate similar to that of room light. Light intensity is a potential source of error.

Introduction As photosynthesis occurs in a leaf, carbon dioxide is incorporated into organic molecules, and oxygen is released. The oxygen can be trapped in bubbles, thus allowing the leaf to float in normal conditions. Thus floating leaves can be directly associated to photosynthesis: as more leaves are observed to float, the reaction has occurred to a greater extent. This is a simple way to quantify the rate of photosynthesis, and allows us to demonstrate the effects of various factors on the rate of photosynthesis. We sought to demonstrate the effect of different wavelengths of light on photosynthesis. 30 leaf discs were punched out for 3 different light treatments: red light, white light, and no light.

In this experiment, we hypothesize that red light will have the same effect as no light on the rate of photosynthesis, which will be substantially less than the effect of white light.

Methodology Three leaf suspensions were created with the given lab procedure, and placed in their respective cup environments. For red and white light, cups were isolated away from room light under a fumehood. Counts for floating discs were taken every minute for a 12 minute period.

Data Time (min) 0 1 2 Red light 0 0 0 White light 0 0 0 No light 0 0 0 Room light* 0 0 0

3 4 5 6 7 8 9 10 11 12

0 0 0 0 0 0 0 0 0 0

1 2 5 6 8 9 9 9 10 10

0 0 0 0 0 0 0 0 0 0

0 0 1 1 2 3 6 6 6 8

Figure 4. Number of floating discs over time. *from previous lab.

Effects of different types of light on the rate of photosynthesis

12 Number of floating discs 10 8 6 4 2 0 0 1 2 3 4 5 6 7 8 9 10 11 12 Time (minutes) Red light White light No light Room light

Figure 5. Lab data graphed.

Results and Discussion Over the course of 15 minutes, neither the red light incubation nor the no light incubation caused any of the leaf discs to float. The white light resulted in a rate of photosynthesis similar to that of room light in the previous lab (where we learned the procedure). This proves our hypothesis. We also note that while the slopes of the white light and room light graphs are approximately equal, white light initiated the reaction faster.

However, we speculated that there may have been a discrepancy between the amount of energy emitted between white and red light. During the incubation period, the three environments were measured with a light intensity sensor. While values fluctuated over time, the white light registered significantly more lumens than the red light or the no light. This may indicate that the change in wavelength of light cannot be directly correlated with the change in rate of photosynthesis. In a future lab, we would attempt to equalize the light intensities in order to isolate wavelength as the only independent variable.