Biol 130 Supplement to Exercise 5 Leaf Disc Experiment (http://www.elbiology.com/labtools/Leafdisk.

html) Hypothesis: Plant leaves require light, a source of CO2, H2O, and chlorophyll in order to carry on photosynthesis. If they are supplied those items, photosynthesis can be observed by the production of O2 gas. 1. Prepare a 0.2% sodium bicarbonate solution for the experiment: 1/8 tsp sodium bicarbonate in 300 ml water. ( OR if stock solution is provided, make the appropriate dilution with water to get 0.2%.)Add 1 drop dilute soap liquid and mix. Do not make suds. Sodium bicarbonate will serve as the source of CO2 in this experiment. Make another solution of 300 ml plain water with 1 drop soap liquid. Prepare 3 beakers, each containing the same amount of liquid (3 cm deep): 2 beakers with the sodium bicarbonate-soap solution, 1 with the water-soap solution*. Label the beakers so you know what each one contains. (do not short-cut here common place to mess up!) 2. Use a hole punch to make the following leaf discs: 30 dark green; 15 light green/white. Avoid large veins in the leaves. Keep the 2 types of leaf discs separate. Carry out step 3 for each set of leaf discs. HOWEVER, for the ONE set of green discs, DO NOT USE sodium bicarbonate-soap solution in step 3; use the water-soap solution only. This set of discs will be tested ONLY in water-soap. 3. Place 15 leaf discs in an opened syringe. Replace the plunger, and depress the plunger until there is very little air left with the leaf discs. Pull up a small (3-5 ml) volume of the sodium bicarbonate-soap solution into the syringe. Block the opening of the syringe with thumb. Pull on the plunger to form a vacuum. Maintain a vacuum for ~10 seconds, gently shaking the discs around at the same time. You should be able to see tiny gas bubbles coming out of the leaf discs. The point is to suck out all the air bubbles inside the leaf discs so that they will sink in the liquid. This vacuum procedure may be repeated 2-3 times until all of the discs are sunken. 4. Place the sunken leaf discs in the beakers prepared in step 1. 15 green leaf discs go into one beaker with bicarbonate-soap solution, and 15 white/light leaf discs go into the second beaker with bicarbonate-soap. The remaining 15 green discs go into the beaker with water-soap. Keep discs away from light until all discs are ready for step 5. 5. Place all 3 beakers under a light source. All 3 beakers should have the same amount of light. For each minute under light, count the number of leaf discs floating. Record data on table.

Biol130 spr11, ML

Number of Leaf Discs Floating min

Green discs, White discs, bicarbonate- bicarbonatesoap soap Green discs, water-soap

(Space for conclusion)

Plot the graph with the y-axis for # of leaf discs floating and the x-axis for Time (in minutes). (Please label these axes properly.) You only need to make a graph for the set(s) of leaf discs that floated. If you plot more than one set of data, please label the data on the graph or provide a key with symbols. The ET50 is the point in time when 50% of the leaf discs are floating. Mark the ET50 for your data on the graph. What does this experiment show? Briefly state a conclusion in the space provided above. For full credit, follow the steps for writing a conclusion from the buffer handout. (Most points lost here. Will put some tips on BB.)

ET50 Graph for Leaf Discs

Biol130 spr11, ML