Output variables – volume of oxygen produced (rate of photosynthesis) is to be measured by finding the volume of oxygen produced in a minute, and thus finding the rate of photosynthesis
Control variables –Light wavelength (colour) – light energy is absorbed by the pigment, chlorophyll, in the leaf. Chlorophyll easily absorbs blue light, in the 400-450 nm range, and also easily absorbs red light, in the 650-700 nm range. However it does not easily absorb green or yellow light, rather it reflects them, decreasing the amount of light absorbed, and therefore the rate of photosynthesis. This can easily be controlled, simply by using the same lamp throughout the experiment.
Carbon dioxide concentration – This can affect the rate of photosynthesis, since if there is too little CO2, it can become the limiting factor, thus impeding the viability of the experiment. In this case, as long as the experiment is done over a short period of time, the amount of carbon dioxide used up by the plant will not be sufficient enough to cause the carbon dioxide concentration to become the limiting factor. If my experiment were to be performed over a longer period of time, for instance 24 hours, I would add a fixed amount of Sodium hydrogen carbonate to the water, thus ensuring a large enough supply of carbon dioxide.
Water availability – water is also required in the photosynthesis reaction, and when it is lacking, the plants’ stomata close to prevent further water loss. This closing of the stomata cells also leads to little carbon dioxide being able to diffuse through. Clearly, in a water plant, like the pondweed, as long as the plant is fully submerged in water at all times, this will not be a problem.
Temperature – Enzymes are used in the photosynthesis reactions of a plant. Therefore, temperature will increase the rate of photosynthesis, until a point at which the enzymes denature. Although performing the experiment at a temperature slightly higher than room temperature, perhaps 25°C, would have a positive effect on the accuracy of the readings I took, as it would reduce the percentage error, by increasing the volumes, I decided that the inaccuracy of maintaining a constant temperature would outweigh any advantages. I am therefore going to perform the experiment at room temperature, checking the temperature frequently, in case the heat given off from the light should slightly raise the temperature, in which case I shall simply refill the beaker with more water after each experiment.
Method
Apparatus list
Desk lamp
Audus apparatus
Canadian pond weed
Knife
Clamp
Pond water
Thermometer
Test-tube
Beaker
Cold water
Stopwatch
Cut a stem of Canadian pondweed of about 3cm in length. Fill a test-tube with pond water, and place it in a clamp, and then in a large beaker of cold water. Connect the end of the pondweed to the Audus apparatus. Insert a thermometer into the beaker, and record the temperature at the beginning and end of each experiment, merely as a precaution against a significant rise in temperature, which is not expected. Set up a lamp at a set distance from the plant, ensuring that this distance is from the filament of the lamp to the actual pondweed, rather than the edge of the beaker. The light intensity was measured in the same way as described in the preliminary experiment, and assumed to be the same at any point at any particular distance. When bubbles are being produced at a steady rate, clear any previous bubbles from the tubing by moving the syringe. Start the stopwatch, and wait for 1 minute. Move the bubbles, which have been collected at the bend in the tubing to the part of the tube with a scale. Find the length of the bubble collected. Repeat for all other readings, and then repeat all readings a second time to get an average result for each distance.
Analysis
My graph was in the form of a best-fit curve. I drew it as a curve rather than a straight line because of the clear pattern of the points. This meant that the rate of photosynthesis increased as the light intensity increased. This was because photosynthesis is a reaction, which needs energy from light to work, so as the amount of energy available from light increased with the rise in light intensity, so did the amount of oxygen produced as a product of photosynthesis.
My graphs showed that the relationship between the light intensity and the rate of photosynthesis was non-linear, as both graphs produced a best-fit curve. However, as I expected in my hypothesis, it does appear that for the very first part of the graph, the increase in rate is in fact proportional to the increase in light intensity (i.e. a straight line) and I can show this by taking some readings from the graph:
Light intensity Rate of photosynthesis
(All increase by the 100 1 (mm/min)
same factor) 150 1.5 (mm/min)
200 2 (mm/min)
From these results, I am able to say that an increase in light intensity does certainly increase the rate of photosynthesis. The gradual decrease in the rate of increase of the rate of photosynthesis (the shallowing of the curve) can be attributed to the other factors limiting the rate of photosynthesis. As light intensity increases, the photosynthetic rate is being limited by certain factors, such as carbon dioxide and temperature. These factors do not immediately limit the rate of photosynthesis, but rather gradually. As light intensity increases further, so the rate of photosynthesis is being limited by other factors more and more, until the rate of photosynthesis is constant, and so is almost certainly limited in full by another factor.
Overall, both graphs and my results support my predictions fully. My idea that the rate of photosynthesis would increase with light intensity was comprehensively backed up by my results. This is because a higher light intensity involves a greater level of light energy, which can then be transferred to a special protein environment designed to convert the energy. Here, the energy of a photon is used to transfer electrons from one chlorophyll pigment to the next. When enough energy has been gathered at a reaction centre, ATP can be synthesised from ADP. The oxygen collected in the experiment is in fact the by-product of this reaction, and so it is clear to see that the more light energy, the more ADP is being converted into ATP and more oxygen is produced as a result.
Evaluation
Although I feel that my experiment was sound overall, I thought there were many points at which the accuracy was not perfect. As I have already stated, my preliminary experiment was not accurate enough to justify being used as my main experiment, mostly due to the fact that I was relying on all the bubbles being the same size, which they clearly weren’t, however many of the smaller inaccuracies also apply to my main experiment.
Firstly, the distance between the light sources and the Canadian Pondweed were not measured to a very high degree of accuracy, especially when you note the fact that the distance should have been measured exactly from the filament of the light bulb to the centre of the plant, and it is possible here to find a percentage error. I estimate that the error could have been up to 0.5cm and I will find the percentage error for the largest and smallest reading using this estimate:
Percentage error = possible inaccuracy
total reading
% error distance
10 5cm
1 50cm
It is clear to see that the percentage error is much less for the larger distances. Although I was not actually using the distances as part of my results, I used them as a marker for where the lamp was placed each time, as I assumed that the light intensity would be the same each time at a particular distance. Therefore, any inaccuracies in measuring the distances, i.e. if a distance was slightly different when doing the actual experiment from the distance at which I earlier measured the light intensity, an error would ensue.
The second major inaccuracy was in measuring the volume of oxygen given off.
When reading the syringe there could have been an error of 0.25mm, and again it is possible to find a percentage error.
% error volume
3.57 7ml
50 0.5ml
For the smallest volumes this is clearly a massive error, and to improve this, it would be necessary to do the readings over a longer period of time, therefore increasing the volumes, and in turn reducing the percentage errors.
Another error would have been due to background light in the vicinity. We tried to reduce this error by closing all blinds in the laboratory, but due to practical reasons, we could not all perform the experiment in a separate room, and we therefore experienced light pollution from other student’s experiments. This would have had a very marginal effect on my results as a whole, but to eliminate this problem completely, it would have been necessary to perform the experiment in a totally dark room.
A further inaccuracy was in the heat generated by the lamp. As I have earlier described, temperature has a very noticeable effect on the rate of photosynthesis, and so any increase in the temperature of the pond water would have had serious effects on the accuracy of my results. To ensure this did not happen, I monitored the temperature of the water before and after every reading, to check that the temperature did in fact not rise. It turned out not to be a problem, as over the short period of time taken by my experimental readings, the temperature did not rise at all. However, if I were to extend the time of my experiment to 5 minutes for each reading for example, which would have the effect of reducing other percentage errors, I would have to find some way of keeping the temperature constant. One way of doing this would be to place a perspex block between the lamp and the plant, which would absorb most of the heat, while allowing the light energy to pass through.
As I mentioned in my planning, carbon dioxide concentration could have been an error in the experiment, however, I feel that due to the short period of time taken, there is very little chance that the concentration would ever have been so low as to have become the limiting factor. Again if I were to carry out the experiment over a longer time period, it would have been necessary to add sodium hydrogen carbonate to the water to increase the carbon dioxide concentrations.
The last inaccuracy, though a small one, was in the time keeping. The main problem here was in when to begin the minute. If for one reading, the minute was started just after one bubble had been produced, and in another reading it was just before, this could have had a negative effect on the accuracy of my results. I therefore ensured that in each case I started the stopwatch just after a bubble had been produced, thus heightening the accuracy.
Overall, I felt that due to the small volumes of oxygen involved, my experiment was not as accurate as it could have been, however I believe it was accurate enough to support and justify my hypotheses. Improvements could have been made as I have stated, mainly by simply increasing the time taken. However, due to practical time constraints in taking the readings for my investigation, and some consequential problems relating to time extension, I could not in fact make these adjustments. The other obvious way of increasing the reliability of my results would be to take many repeat readings and find an average.
To extend my enquiries into the rate of photosynthesis, I could perhaps try to link in some of the other limiting factors to the same experiment, as well as investigating them in their own right. It could also be interesting to explore the effects of coloured lights on the rate of photosynthesis, which could lead to the question of whether or not other types of light, such as fluorescent lights or halogen lights, would have a different effect on the rate of photosynthesis.
Evaluation:
Overall, I would state the experiment as a success since my predictions were supported by my results. This is important in reflecting success only if my prediction was sensible and logical. Just as important is where the experiment was not a success and why. This photosynthesis investigation was probably not performed as accurately as it could have been due to some controllable and uncontrollable conditions. Some mistakes can be corrected.
While performing the experiment, the piece of pondweed did not photosynthesize at a steady rate, even when the distance from the plant to the light source was kept a constant. The second reading at 0 cm was far greater than the first reading at 0 cm. While the number of oxygen bubbles was being recorded, the rate at which the plant was photosynthesizing had increased several times. This may be due to the poor circulation of sodium hydrogen carbonate at the beginning of the experiment. Carbon dioxide may have initially limited the rate of photosynthesis. The readings at 0 cm and 5 cm were repeated many times until the rate of photosynthesis had begun to settle. From then on, there were no more similar problems during the experiment. To make sure that the there
The negative effects from this problem may be inaccurate data for some readings. These would show up on my graph. However, there seemed to be few anomalies than was expected when the experiment was being performed. Almost all readings were in correlation with each other and all of the anomalies were in the high photosynthetic rate end of the results. This was when the distance from plant to light source was 0 cm or only 5 cm.
A large factor in determining data accuracy is the amount of human error during experiments. The rate at which oxygen bubbles were being produced by my plant was so high that I found it difficult to count the amount of bubbles. I estimate a margin of error of at least 3 bubbles for each reading taken. To improve the accuracy of the results, the readings would have to be taken several more times. The entire experiment could have been performed again, and the new results could be combined if the same plant is used. But the photosynthetic rate of the same piece of pond weed would eventually decrease over time anyway. Repetitions would, however, improve the overall reliability of the results.
There are quite a few factors that could affect the results of my experiment. Some of these are variables that were mentioned earlier and could not be controlled, or they were variables that were not initially considered.
While performing the experiment, some of the oxygen produced from photosynthesis may have dissolved into the water. Some oxygen may have even been used by micro-organisms living on the pond weed. The amount of oxygen dissolved or used by microbes is probably insignificant to my results since the degree of accuracy at which I measured was not high enough. Some oxygen is also used during the respiration of the plant. But since only bubbles were counted, the volume of bubbles was not as important. But to volume of oxygen produced is important, since it was volume in terms of bubbles that were measured. As the rate of photosynthesis decreased due to a decrease in light intensity, the size of the bubbles produced also became smaller. This change in bubble size was no accounted for when the results were analyzed. For a more accurate analysis of the collected data, volume should have been measured instead of bubble quantity since the size of bubbles can vary. Using a capillary tube in place of the test tube so that the volume of each bubble could have been measured could have done this.
During the high intensities I had experienced counting difficulties of the bubbles being produced. There are also factors affecting accuracy at low light intensities. With low light intensity, the pond weed receives some light energy from background light such as sunlight seeping through curtains or the light from the lamp of another student's experiment. To eliminate most all background light, the experiment must be performed in a completely dark room. Even then, some of the light from the lamp in my experiment would reflect of the table and reach the plant though this amount of light is probably insignificant in affecting the rate of photosynthesis.
Temperature was also another factor that was controlled by the lamp being used. Even though a glass block was used in front of the lamp to prevent some heat from reaching the plant, not all the heat can be blocked. The extra heat, however, did not affect the temperature of the water, which stayed at between 290 and 300 C.
The method of the experiment could probably also be improved to obtain more reliable results. As already mentioned, the a capillary tube should be used in place of a test tube to accurately measure the volume of the oxygen produced. Due to the high rates of photosynthesis of the pond weed, readings should be taken within shorter time periods. I had originally chosen to count the number of bubbles in one minute but this produced miscounts in the readings. If during a repeated experiment, counting bubbles is still used, there is a smaller chance for human error when counting within a smaller time frame. If the capillary tube option was to be chosen, volume should be measured for a smaller time frame to reduce the overall time to complete the experiment. Also, during high rates of photosynthesis, it would still be difficult and impractical to measure the volume of oxygen produced for a long duration.
Due to the nature and convenience of the experiment, it could be easily modified to investigate another variable of photosynthesis. Since sodium hydrogen carbonate (NaHCO3) is used to provide the pondweed with carbon dioxide. Performing the experiment with different volumes of NaHCO3 could vary the amount of CO2. The plant would be kept at a constant distance from the lamp and a constant volume of water would be added to the sodium hydrogen carbonate. Another experiment using almost identical apparatus would be to vary the color of the light the plant absorbs. Using translucent color filters in front the lamps could vary this. Since light wave length has already been identified as a variable of photosynthesis, it would be interesting to actually test it. The only problem of this experiment is that there is no way to define or "measure" the color of light. Wave length would be a solution but this cannot be measured with available equipment. We only have a general idea of how to class colors. Because of this, the colored light experiment should not be taken as seriously as light intensity or carbon dioxide.