MA 61 Precalculus for Biology Majors Lab III: Modeling Data Using Regression – Photosynthesis The primary source of energy for nearly all life is the Sun. The energy in sunlight is introduced into the biosphere by a process known as photosynthesis, which occurs in plants, algae and some types of bacteria. Photosynthesis can be defined as the physico-chemical process by which photosynthetic organisms use light energy to drive the synthesis of organic compounds. The photosynthetic process depends on a set of complex protein molecules that are located in and around a highly organized membrane. Through a series of energy transducing reactions, the photosynthetic machinery transforms light energy into a stable form that can last for hundreds of millions of years. 6CO2 + 6H2O + sunlight ---> 6O 2 + C6 H12O 6 or... carbon dioxide + water + sunlight ---> oxygen + carbohydrate (sugar) This lab focuses on showing how mathematical models can be used to study the structure of the photosynthetic machinery and the reactions essential for transforming light energy into chemical energy. To produce photosynthetic reactions plants absorb energy from sunlight. A graph of the visible spectrum of light at different wavelengths and colors is provided below: Introductory Task: Understand the Problem to be Modeled Open and view the first 20 or so slides of the Photosynthesis_self_study_exercise PowerPoint slide show presentation in the Lab 3 folder on the CD. This is a long and thorough description of photosynthesis that you may wish to view in its entirety at a later time for your biology class. Lab Assignment Part 1: Interpreting Graphs Task I: Plotting and Interpreting an Absorbance Curve Sample Absorbance versus Wavelength data is provided in an Excel spreadsheet (Photosynthesis.xls). Open the Photosynthesis.xls Excel spreadsheet and plot the Absorbance data using what you have previously learned with the Chart Wizard and then answer the following questions: 1. What wavelengths give you the higher absorbance levels? What color/colors are these wavelengths associated with? 2. What wavelengths give you the lower absorbance levels? What color/colors are these wavelengths associated with? 1 3. What are plants green in color? Lab Assignment Part 2: Constructing a Model I How do we measure photosynthesis? Well, obviously from the chemical reaction above, if we can measure the amount of oxygen produced, then we also know how much carbohydrates or sugars are also produced. A measure of how much oxygen is produced and, therefore the level of photosynthesis, can be determined from an O2 evolution curve. O2 evolution curves A light response curve is an example of something called a "second-order" graph, since the rate of photosynthesis first must be determined by plotting O2 evolution versus time at different light levels. The rates calculated from each O2 evolution curve are then plotted against light level. For example, the figure below shows O2 evolution curves when photosynthesis was allowed to occur in the presence of two different light levels. Notice that there is a time period (called the lag period) between the time the lights are turned on and when photosynthesis reaches steady state. Task II: Modeling Oxygen Evolution Open the Photosynthesis.xls spreadsheet and plot the five sets of oxygen production data at the various light intensities, i.e. 50, 100, 200, 300 and 400 m/(m2 s). Find the “best-fitting” curve and write down the associated regression curve equation. O2 evolution rates The rate of photosynthesis is calculated from the slope of the linear part of the O2 evolution curve. This figure shows regression lines drawn through the steady-state regions of the O2 evolution curves. 2 In experimental setups, O2 and light levels are monitored automatically by sensors. The O2 sensor is calibrated to measure "partial pressure" of O2-- the amount of O2 relative to other gases in the atmosphere. Since the units of partial pressure are %O2, the slope of the line will have units of %O2 / min. This value can be used to calculate a photosynthesis rate with units of 'umole O2 / m2 / min' by factoring in leaf area and using proper conversion factors. The Photosynthesis Light Response Curve When photosynthesis rates from several experiments are plotted against light intensity, the result is a photosynthesis light response curve, such as the one shown below. Interpreting the photosynthesis light response curve Different plants (even different leaves on the same plant) show differences in the shape of their light response curves, which reveals characteristics of the underlying photosynthesis processes including the light-dependent and light-independent reactions, the efficiency at which light is utilized by photosynthesis, and even the rate of O2 uptake. The response curve can be divided into two phases. Notice that under low-light levels, the rate of photosynthesis increases as the irradiance level is increased. 3 The Light Saturation Point At a particular light intensity, the so-called "light saturation point", the rate of O2 evolution levels off. Any further increase in the amount of light striking the leaf does not cause an increase in the rate of photosynthesis-the amount of light is said to be 'saturating' for the photosynthetic process. Task III: Modeling a Light Response Curve From the graphs in Task II above, you can determine the photosynthetic rate for each of the five intensities by taking the slope of each line. Once you have done this, prepare a second graph plotting photosynthetic rate versus light intensity. Use this curve to approximate the Light Saturation Point. The Light Compensation Point Notice that when extrapolated down, the light response curve does not pass through the origin of the graph. The light value on the X-axis through which the line passes is called the "light compensation point." As you can see, at light levels below this, there is no net O2 evolution. Do not misinterpret this to mean that photosynthesis does not occur at light levels below this point, there is a better explanation. 4 You should note that this line does not pass through the origin. Instead, it crosses the x-axis at what is called the photosynthetic compensation point. The photosynthetic compensation point is the intensity of light required for no net carbon assimilation, meaning that the CO2 consumed by photosynthesis is exactly the same amount produced by respiration. At light intensities above the photosynthetic compensation point, the plant is taking in more carbon that it is producing as a waste product by its metabolism. At light intensities below the photosynthetic compensation point, the plant is actually losing carbon! Task IV: Determine the Light Compensation Point In this task we will determine the photosynthetic compensation point. Use the light Response Curve from Task III above and find the appropriate equation of the line for this curve (see figure above). Then set y = 0 in the resulting equation and solve for x. The x value is the compensation point. Note: There is no such thing as a negative compensation point (this would imply that the plant is gaining energy from photosynthesis in total darkness). This lab sometimes doesn't work for one reason or another and when you solve for x in the above equation, you may find that the compensation point is negative. You should interpret this as an illogical result and simply state that you cannot determine the compensation point with your dataset and recommend that you perform the experiment again Photosynthetic Efficiency The slope of the linear phase of the response curve is a measure of "photosynthetic efficiency" -- how efficiently solar energy is converted into chemical energy. 5 The photosynthesis apparatus we use is not designed to directly measure the amount of light absorbed by the leaf. Nevertheless, the light response curves that the class produces do allow comparisons in photosynthetic efficiency between plants. Task V: Determine Photosynthetic Efficiency Use the models created above to determine the photosynthetic efficiency of the data set. 6