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The Effect of Boiling on the Rate of Aerobic Respiration of Germinating Peas

Abstract
Aerobic respiration is an enzyme mediated process during which glucose is used to make ATP to fuel the cells energy needs. Germination is one of such processes that make use of aerobic respiration and because certain factors can affect aerobic respiration, it would also affect germination. In this experiment, we examined the effects of boiling on the rate of aerobic respiration of germinating peas. Unboiled germinating peas were found to have a more rapid rate of aerobic respiration than boiled non-germinating peas. Boiling therefore slows down and destroys the ability of peas to germinate or undergo aerobic respiration.

Introduction
Living organisms undergo cellular respiration during which organic food molecules are oxidized to synthesize ATP used to drive the metabolic reactions necessary to maintain the organisms physical integrity and to support all its activities. This is achieved by either aerobic (in the presence of oxygen) or anaerobic (in the absence of oxygen) respiration. Aerobic respiration is a more efficient process as up to 30% of the energy in glucose makes its way to ATP whereas only about 2% of the energy available in glucose is usable by the cell through anaerobic respiration (1). The chemical reaction for aerobic respiration is: C6 H12 O6 + 6 O2 + 6 H2 O CO2 + 12 H2 O + energy to make 36 38 ATP

Germinating peas breakdown their starch stores to produce glucose which is then used to make ATP aerobically so that shoots and seedlings can be made. This high aerobic respiration rate of germinating peas can be affected when they are exposed to different conditions. In this experiment, we examine the effects of boiling on the rate of aerobic respiration of germinating peas.

Materials and Methods

A large number of peas had been previously soaked in water for 48 hours to initiate germination, after which one set of the germinating peas were boiled and while the other set unboiled. A small ball of absorbent cotton wool was added to the bottom of three labeled test tubes and then 4 drops of 15% potassium hydroxide (KOH) were added directly to the absorbent cotton in each tube. Another layer of non-absorbent cotton ball was used to cover the moistened cotton layer at the bottom of each tube. Eight regular (unboiled) germinating peas were added to one tube and 8 boiled germinating peas were added to another tube. No peas were added to the third tube which served as the thermobarometer, a control for temperature and pressure in the laboratory environment. A second ball of non-absorbent cotton was again added into all three tubes so as to cover the peas. This layer of non-absorbent cotton ball was also moistened with 2 drops 15% KOH. To each of the 3 tubes, a dry stopper-syringe/pipette assembly in was inserted to for a respirometer. By means of a 1-ml Pasteur pipette, a small drop of marker fluid was added to the end of each pipette to forma a closed system. The position of the drop was then adjusted to the zero mark by gently withdrawing the plunger on the syringe. Each respirometer was left to equilibrate for 15 and then 5 minutes, after which times, the position of the marker fluid was adjusted to the zero mark. After the last 5 minutes were over, the volume (in ml) moved by the marker drop, due to oxygen being consumed was measured every 3 minute, for 15 minutes. The measured volume of oxygen consumed as cellular respiration took place was corrected for any by atmospheric temperature or pressure by subtracting the values obtained from the thermobarometer. The rate of oxygen consumption (rate of aerobic respiration) for both boiled and unboiled germinating peas was determined by plotting a graph of volume-of-oxygen-consumed vs time and a best fit line was drawn through the data points. The mass specific rate of oxygen consumption was also obtained by dividing the slope by the mass of the peas used. These values were also corrected for standard temperature and pressure.

Results
In order to determine the effect of boiling on the rate of aerobic respiration, we measured the amount of oxygen consumed by the germinating peas as they respire and compared it with that of boiled peas. Since carbon dioxide (CO2) is also released during respiration, the use of KOH to remove CO2 (forming potassium carbonate) during the process ensured that the measured volume of gas in the respirometer would be due to the consumption of Oxygen. Figure 1 shows that the rate of oxygen consumption (rate of aerobic reaction) of the unboiled germinating seeds was higher (slope = 0.22ml/min) than that for the unboiled peas (slope = 0.05ml/min).

4 Volume of Oxygen consumed (ml) 3.5 3 2.5 2 1.5 1 0.5 0 0 2 4 6 8 Time (min) 10 12 14 16 y = 0.0514x + 0.0476 Unboiled germinating peas Boiled germinating peas y = 0.2181x + 0.1143

Figure 1. Rate of oxygen consumption of boiled and unboiled germinating peas. As cellular respiration occurs, more oxygen is consumed, with more oxygen being consumed (high rate of aerobic respiration) by the unboiled germinating seeds compared to the boiled seeds which virtually respired (lower rate of oxygen consumption compared to unboiled peas).

At standard temperature and pressure, the rate of aerobic respiration of the boiled peas is almost negligible compared to that of the germinating peas. Table1: Effect of Boiling on the rate of respiration in peas Peas Slope(ml/min) Seed Mass (g) Mass-specific rate of O2 consumption(ml/min/g) @ rtp (298K, 761 Torr) 0.040 0.008 Mass-specific rate of O2 consumption(ml/min/g) @stp (273K, 760 Torr) 0.037 0.007

Unboiled, germinating Boiled

0.22 0.05

5.48 5.58

Discussion and Conclusion

It is expected that only actively respiring/germinating peas would show an increase in respiratory rate. This was the case with the unboiled peas which had a higher rate of oxygen consumption as aerobic respiration with germination. Cellular respiration is an enzyme-mediated process involving several enzymes that catalyze the reactions of the glycolysis and the Krebs cycle. Boiling irreversibly altered the 3D structures of enzymes such that they become less functional and lose their catalytic activity (2). Thus the enzymes in the boiled peas would have lost their enzymatic activity due to boiling and these peas would not be able to undergo aerobic respiration. This suggests that peas will not germinate or even grow in harsh climatic conditions with very high temperatures. However, in our experiment, we noticed that the rate of oxygen consumption for the boiled peas was not zero as expected. This would have come about as a result of erroneous volumes-ofoxygen-consumed readings on the respirometer due to the fact that the stopper did not tightly seal the respirator and a close system was not obtained. False volume readings on the boiled peas respirator would also have resulted from disturbing the respirometer when taking readings such that the marker drop on pipette moved from the zero mark, giving a false volume reading. Nevertheless, the rate of respiration for the boiled peas were very small and negligible compared to those for the unboiled germinating peas.

Our experiment therefore shows that unboiled germinating peas carried out a more rapid process of cellular respiration than boiled non-germinating peas. Boiling therefore slows down and destroys the ability of peas to germinate or undergo aerobic respiration.

Literature Cited
1) Dolphin, W. D (2011) Biological Investigations: Form, Function, Diversity, and Process. 9th edition. WCB/McGraw-Hill, Boston, MA 2) Campbell, N. A., J. B. Reece, and L. G. Mitchell.(2012) Biology. 9th edition. Pearson Benjamin/Cummings, Menlo Park, CA, USA. Pg. 151-155.