Biology Lab Photosynthesis

1. Is there evidence that chloroplasts were able to reduce DPIP in this experiment? Explain.

Yes, DPIP is reduced by the chloroplasts in the solution. DPIP replaces NADP as an electron
acceptor when hydrogen is separated from H20 as a result of the chlorophyll in the chloroplasts. When
the solutions with DPIP in them were exposed to light, the transmittance percentage had an increasing
tendency, meaning more light is able to shine through the solution. The dark color from the DPIP
became much lighter and more see-through as chloroplasts used it in place of NADP, reducing it and
therefore changing the color.

2. Were chloroplasts able to reduce DPIP when kept in the dark? Explain.
DPIP is not reduced in the dark. Chloroplasts cannot oxidize H20 if light cannot provide the
photons to energize electrons. Electrons must be excited to be oxidized from H20, and if they cannot
be separated from water by the chlorophyll, no electrons can be freed to reduce DPIP.

3. Were boiled chloroplasts able to reduce DPIP? Explain.

No, boiled chloroplasts did not and can not reduce DPIP. The proteins and enzymes necessary
for photosynthesis, for example the catalyzer RuBisCO in the Calvin cycle, become denatured when
heated, making oxidation of electrons impossible. Proteins are necessary in the electron transport
chain, if they cannot “pass down” electrons, photosynthesis is completely halted because ATP cannot
be produced. If no electrons can be freed by oxidation, there are no free electrons to reduce DPIP.

4. What conclusions can you make about the photosynthetic activity of spinach?
Photosynthesis occurs at a faster rate when the chloroplasts found in spinach cells are exposed
to more light for longer periods of times. The amount of light and exposure to it directly effects the
rate at which DPIP was used in the experiment. DPIP serves as an electron acceptor, reducing the
molecule and serving as a replacement to NADP, which is the normal electron acceptor during
photosynthesis. The more DPIP that was used, or the clearer the solution got, the more photosynthetic
activity that was going. Chloroplasts that are boiled denature the enzymes and proteins found in the
electron transport chain, crippling the entire processes and making it impossible to conduct.