Laboratory Exercise No.

PHOTOSYNTHESIS, RESPIRATION AND TRANSPIRATION

Maria Karina P. Degoma Crop Science 1 B-1L

July 22, 2013

ABSTRACT Photosynthesis is a process used by plants and other organisms to convert the light energy captured from the sun into chemical energy that can be used to fuel the organism's activities. It also splits water to liberate O2 and fixes CO2 into sugar. Light intensity is one of the main factors affecting rate of photosynthesis, respiration, and transpiration. In effect, higher light intensities make more energy available for photosynthesis to take place. An experiment was conducted showing that under different light intensities the photosynthetic rate of plant varies. Gross photosynthesis is the sum of the dry weight and amount of photosynthates utilized for respiration. The results of the experiment showed that gross photosynthesis is higher when the plant is exposed to full sunlight compared to partial shade. The rate of transpiration was also determined under different conditions: still air and low light, still air and strong light, with fanning and low light, and with fanning and strong light. the rate of transpiration is lowest in condition with still air and low light, and highest in condition with fanning and strong light. As light intensity increases, the rate of the light-dependent reaction increases as well, therefore photosynthesis increases proportionally, as well as respiration and transpiration.

INTRODUCTION One of the two plant physiological processes which occur primarily in leaves is photosynthesis. It is responsible for the maintenance of all life on the planet, either plant or animal. Photosynthesis enables plant organisms to produce food needs. It is the process of capturing light energy converting it to sugar energy, in the presence of chlorophyll using carbon dioxide and water. The net result is where light energy (radiant energy) absorbed by pigments inside the chloroplast is converted into chemical energy in the form of fixed carbon compounds (carbohydrates). The energy absorbed becomes the fuel utilized to manufacture glucose, or sugar molecules. Photosynthesis occurs when plant leaves absorb the energy contained in light. Energy from light can cause an electron of a chlorophyll molecule to dislodge from its orbit and move to a higher orbit, getting excited. The electron may then immediately fall back to its original orbit, releasing energy which causes an emission of light (fluorescence). Plants are typically most responsive to light that falls within the range of blue and red light. Light color represents the different energy frequencies contained within the visible light portion of the electromagnetic spectrum. Red color contains the lowest energy frequencies, while those toward the blue-violet end carry the highest frequencies. In effect, light intensity can vary depending on the type of color used as well as the wattage amount contained in a bulb. The energy intensity provided by red light stimulates overall growth throughout the plant body, while blue light helps to regulate growth rates within the leaves, stems and flowers. The light intensity range for most plants falls between 1,200 and 2,000 lumens. As the light intensity rises, the number of light photons

hitting the chlorophyll pigments increases. Thus, increases the rate of photosynthesis. The whole process is so effective that in optimum conditions 90 percent of light is absorbed.

The structural unit of photosynthesis is the thylakoid . Both photosynthetic prokaryotes and eukaryotes have these flattened sacs/vesicles containing photosynthetic chemicals. Only eukaryotes have chloroplasts with a surrounding membrane. Thylakoids are stacked like pancakes in stacks known collectively as grana. The areas between grana are referred to as stroma. Stroma are the fluid matrix of the chloroplast.
(http://www.emc.maricopa.edu/faculty/farabee/biobk/BiobookglossG.html)

A pigment is any substance that absorbs light. The color of the pigment comes from the wavelengths that were reflected. Light that passes through or is reflected by a leaf cannot contribute energy to the photosynthetic process; only light that is absorbed by the leaf can contribute to photosynthesis (Schooley, J. 1997). Chlorophyll, the green pigment common to all photosynthetic cells, absorbs all wavelengths of visible light except green which is reflected, and is the color detected by our eyes. Black pigments are the one that absorbs all of the wavelengths that strike them. Pigments that reflect all or almost all of the energy striking them are the white pigments/lighter colors.. A remarkable aspect of plant metabolism is that during photosynthesis, plants use CO2 and water to make a single simple organic compound, 3-phosphoglyceraldehyde. Then by altering it chemically with various enzymes, the plants use 3phosphoglyceraldehyde to construct numerous types of carbohydrates and lipids. By adding simple compounds of nitrogen, phosphorus, sulfur, and so on, all derived from the soil, the plants use the 3-phosphoglyceraldehyde and its derivatives to make amino acids and proteins, nucleotides and nucleic acids, and every other compound in them. Actually,

to build its entire body and carry out its metabolism, a green plant needs only sunlight, carbon dioxide, water, and a few mineral elements (Mauseth, J.D. 1998) . Light from plants point of view has three important properties: duration, quality, and quantity. Duration of light refers to the sunlight available in number of hours per day. Quality of light refers to the colors or wavelengths that it contains. Quantity of light refers to light intensity or brightness. Plants growing in the shade receive not only dim light, but light depleted of red and blue while plants that grows on slopes that face the sun or at the equator receives intense light. Higher light intensities make more energy available for photosynthesis to take place. As the light intensity rises, the number of light photons hitting the chlorophyll pigments increases. In return, this increases the rate of photosynthesis. In 1905, when investigating the factors affecting the rate of photosynthesis, Blackmann formulated the Law of limiting factors. This states that the rate of a physiological process will be limited by the factor which is in shortest supply. Any change in the level of a limiting factor will affect the rate of reaction. As light intensity increases, the rate of photosynthesis will increase as long as other factors are in adequate supply. As the rate increases, eventually another factor will come into short supply. Plant leaves contain chloroplasts which are light-absorbing cells. Chloroplasts are the green plastids where photosynthesis occurs. It contains a pigment material called chlorophyll. Chlorophyll responds to different intensities of light, which comes from availability of light. To power the synthesis of ATP, light energy is absorbed by chlorophyll a molecule. In the interior of the thylakoid membrane, water is split into oxygen gas that is given off as a by-product of photosynthesis and Hydrogen atoms.

+6

+ light
(glucose)

6
(oxygen)

Objectives At the end of the exercise, the student was able to: 1. Discuss and relate the role of photosynthesis and photosynthetic organs to yield formation; 2. Discuss the principles, applicability, and limitations of the different methods used in estimating photosynthesis, respiration and transpiration; 3. Estimate the net photosynthesis and respiration in corn plant; and 4. Estimate the transpiration of a herbaceous leafy shoot.

METHODOLOGY Materials The materials used in this experiment were: two Corn plants at silk stage, scissors/knives, paper bags, plastic bags, foot ruler, shipping tags, marking pens, triple beam balance(with sensitivity= 0.1 g), and drying oven. Procedure a. Photosynthesis, Respiration, and Yield The laboratory class was divided into three groups. Then, each group randomly selected six uniform corn plants at the silk or blister stage. It is necessary for the groups to select plants of the same maturity, in this experiment the corn plant chosen was on its blister stage. In this experiment, two light treatments with three plants per treatment were considered. In the first treatment, the corn plants were grown under uninterrupted sunlight (unshaded). In the second treatment, a net house was used to provide corn plants with artificial shade (shaded). Shipping tags were used to label each plant accordingly. The shipping tags contained the following information: laboratory section, group number, treatment, and the plant number. According to the original procedure, it is important to maintain only one ear per plant and that additional ears that emerged from the test plants should be removed. However, it was not followed in this experiment. Instead, all the ears were collected and the average weight of the ears for each corn was obtained.

After 4 weeks of tagging, the tagged plants were uprooted in such a way that the roots were recovered as well. The adhering soil particles from the root system were removed then the plants samples were taken to the laboratory room for preparation. The ears were removed from the plant, dehusked, arranged according to treatments and pictures of the ears were taken. The fresh weight was determined, as well as the dimensions of the ear. The ear length was taken from the base up to the tip of the ear, while the ear width was taken at the middle portion of the ear. The number of kernels was also determined, by counting the number of rows and the number of kernels in each row. Each ears were divided into smaller pieces and was then placed into a separate and properly-labeled paper bags. The samples were dried in an oven with a temperature of over 80 degrees Celsius. After one week, each sample was weighed using the triple beam balance. The data is shown in Table 3.1. b. Determination of Transpiration Leafy branch/shoot was mounted in the photometer. The end of the branch/shoot with a woody stem about the size of a pencil was inserted through a hole of the rubber stopper, making it snuggly but not compressed. The cut end of the specimen was held under water, and about one inch was cut off. The stem was then left immersed in the water. The photometer was then filled with water through the reservoir. The pinch cock was then closed. The stem was moved quickly to the well and inserted the stopper carefully. The stem was allowed to transpire with the end of the capillary tube exposed to the air. After the bubble has entered the capillary, the capillary end was immersed into the auxillary water source. The rate of transpiration was determined by determining the distance travelled by the bubble per unit time interval.

RESULTS AND DISCUSSION Photosynthesis, respiration and transpiration can be measured in many ways as these processes involve the production of oxygen, the uptake of carbon dioxide and an increase in biomass. Plants release oxygen bubbles during photosynthesis and so these can be collected and measured. Photosynthesis can also be measured through an increase in biomass. Light intensity, temperature, amount of water uptake and air movement are just few among the factors that affect these processes. The data collected from the experiment showed the relationship of light intensities and air movement with photosynthesis, respiration and transpiration.

Table 3.1 Characteristics of corn ears harvested from plants grown under full sunlight and partial shade.

PARAMETER Ear length(cm) Ear width(cm) No. of kernel rows No. of kernels/row Ear fresh weight Ear dry weight Other Observations

1 20.2 4.8 14 27 224 168.8

LIGHT TREATMENT Full Sunlight Partial Shade 2 3 Ave 1 2 Ave 17.2 7.1 14.83333 11.3 17 14.15 4.6 3.4 4.266667 3.5 4 3.75 14 12 13.33333 12 12 12 26 10 21 18 15 16.5 204 41 156.3333 74 112.3 93.15 167.8 12.5 116.3667 42.6 72.2 57.4 smaller ears, thinner, has few number of kernels

plant has three ears, bigger and healthier-looking

Table 3.2 Gross photosynthesis, net photosynthesis, and respiration of corn plants grown under full sunlight and partial shade.

PARAMETER Fresh weight (g) Net Photosynthesis (g) Gross photosynthesis (g) Respiration (g)

1 696.8 405.3 604.93 199.63

LIGHT TREATMENT Full Sunlight Partial Shade 2 Average 1 2 Average 558.8 627.8 339.8 446.9 393.35 220.3 312.8 170.1 161.5 165.8 328.81 466.87 253.88 241.04 247.46 108.51 154.07 83.78 79.54 81.66

To calculate the rate of photosynthesis there are few key methods to be considered: measuring the uptake of carbon dioxide, measuring the production of oxygen, and measuring the increase in dry mass. In this experiment there are only three methods used, measuring the oxygen production and measuring the dry mass. Dry mass is often monitored by the technique of 'serial harvests' where several plants are harvested, dried to constant weight and weighed - this is repeated over the duration of the experiment. Harvesting several plants and recording how much mass they have accumulated, an accurate measure of the surplus photosynthesis over and above the respiration that has taken place was obtained. As with most methods, several plants were needed to have replicate measurements and to obtain an average of the results. In the first part of the experiment, the corn ears of each plant under full sunlight and under partial shade was compared. Based on the observations in Table 3.1, the corn plant under full sunlight has three corn ears, which are bigger and healthier-looking compared to the ear of corn plant under partial shade. In the second part of the experiment, the gross photosynthesis, net photosynthesis and respiration was estimated. The apparent or net photosynthesis is approximately equal to the dry weight (weight after oven-dried) of the plant or plant organ. It is about 67% of

the gross photosynthesis, while respiration is about 33%. The gross photosynthesis is the sum of net photosynthesis and amount of photosynthates utilized for respiration. The estimated value for gross photosynthesis was obtained by first dividing the dry weight or net photosynthesis by 0.67, and then the result was multiplied to 0.33 to obtain a value for respiration. The two results were added and noted as the estimated value for gross photosynthesis. In determining the estimated gross photosynthesis and respiration, two conditions were considered: the plant exposed to full sunlight and the other under partial shade. The results showed that the plant exposed to full sunlight obtained a higher value for all the parameters. Light intensity is one of the factors that affect photosynthesis and respiration. The relationship between light intensity and these two processes is directly proportional. As the light intensity increase, the rate of photosynthesis and respiration as well, thus resulting to higher yield.
Table 3.3. Determining the effect of varying the combination of air and light on the rate of transpiration.

CONDITION still air + low sunlight still air + strong sunlight with fanning + low light with fanning + strong light

Trial 1 0.1 0.2 0 0.3

TRANSPIRATION RATE Trial 2 Trial 3 Average 0 0 0.033333 0.3 0.3 0.266667 0.3 0.3 0.2 0.3 0.3 0.3

The third part of the experiment was designed to measure transpiration rates in a particular species of herbaceous plant. The plant was exposed to different conditions. In still air with low sunlight, the average transpiration rate is 0.03 ml/minute. In still air with strong sunlight the average rate was 0.27 ml/minute. The other condition where there is

low light with fanning, the rate was 0.2 ml/minute, while in strong light with fanning the transpiration rate increased to 0.3 ml/minute. The results showed that the transpiration rate is the least in conditions where there is still air and low light. As light intensity increases, the rate of transpiration also increases. This was observed in the second condition where there is still air and increased light. It was shown that from 0.03 ml/minute in low light condition, the transpiration rate increased up to 0.27 ml/minute in strong light conditions. In the third condition, the effect of air movement was also observed. As soon as the fan was introduced to the leafy shoot the average transpiration rate becomes 0.2 ml/minute. The air caused more water to evaporate from the leaf surfaces, so more water was lost and the rate of transpiration increased. Thus, compared to the still air and low light condition, the transpiration rate is relatively higher when air movement is changed given that the light intensity remains the same. The difference in transpiration rate if either light or air movement is present can be compared by looking at the results in the second and third condition. It was clear that there is a slight deviation on the results observed. The transpiration rate in the second condition is higher compared to the third condition. Accordingly, light intensity affects transpiration rate more compared to air movement. In conditions where the factors that affect the rate of transpiration were at its maximum, it was expected that during this conditions transpiration rate was the highest. This was observed in the fourth condition, with fanning and strong light. The environmental factors that greatly affect the rate of transpiration are relative humidity, air movements, temperature, solar radiation, and soil conditions. In any given situation, any one of these factors may become a limiting factor. In other words, the

factors that directly affect the rate at which transpiration can take place masking the effects of the other factors. Temperature increase brought on by strong light and air movement by using a fan is determined to be the factors that accelerated the rate of transpiration.

SUMMARY AND CONCLUSION The effect of light intensity in photosynthesis and respiration was observed by comparing the corn ears of the plants under different conditions and getting the dry weight or net photosynthesis of corn plant and determining the gross photosynthesis. The corn plants under full sunlight were observed to have healthier and bigger ears, compared to the plant under partial shade. In terms of estimated gross photosynthesis, net photosynthesis and amount of photosynthates used in respiration, the plant under the condition of full sunlight showed higher values and better results. In the next part, the experiment performed includes four conditions: still air + low light, still air + strong light, with fanning + low light and with fanning + strong light. Using a leafy shoot, time in seconds wherein a bubble along the capillary tube of a photometer travels from a measured mark to another is obtained. Since the major division of the capillary tube represents a volume of 0.01, the transpiration rate was obtained by dividing 0.01 by the time in seconds the bubble travels (see equation1). The result shows that the rate of photosynthesis in still air + low light is 0.03 ml/minute, 0.27 ml/minute in still air + strong light, 0.20 ml/minute in fanning + low light, and 0.30 ml/minute in fanning + strong light. Transpiration rate = 0.01mL/time in seconds Eq. 1

Light intensity has to do with the amount of light energy made available to a plant, which can vary according to color and the actual strength of the light. It has a very large effect on the rate of transpiration. Low rate of transpiration was observed under still air and low light while a high rate was monitored under with fanning and strong light. In

other words, the more light available the faster the transpiration. Rate of transpiration accelerates as light intensity increases, which also means increase in temperature.

REFERENCES http://www.saps.org.uk/secondary/teaching-resources/157-measuring-the-rate-ofphotosynthesis http://www.life.illinois.edu/crofts/bioph354/lect20.html http://www.emc.maricopa.edu/faculty/farabee/biobk/biobookps.html Mauseth, J.D. 1998. Botany: An Introduction to Plant Biology. pp. 262-294. Schooley, J. 1997. Introduction to Botany. pp. 89-100.