CHAPTER I PRELIMINARY

A. Background

Respiration organic food molecules are oxidized and these exergonic oxidation reactions are coupled with the synthesis of ATP, an endergonic reaction. The ATP is then used to drive the metabolic reactions necessary to maintain the organisms physical integrity and to support all its other activities. The cytoplasm of all cells contains the enzymes needed in the ancient central pathway of glycolysis, in which glucose is oxidized to pyruvate in the absence of oxygen. The energy released in this process is used to generate ATP directly by substrate level phosphorylation, in which phosphate groups are transferred directly from organic substrates to ADP. In many organisms, respiration can occur under anaerobic conditions where no oxygen is present. Many bacteria, yeast, and animals ferment glucose, producing lactate or ethanol. During fermentation reactions, hydrogens are removed from glucose, passed to the electron carrier NAD+ (forming NADH), and then on to pyruvic acid (the end product of glycolysis), converting it to lactate or ethanol. Concurrently, the NADH is oxidized to NAD+, reconstituting the NAD+ pool required for glycolysis. Fermentation allows cells to make ATP in the absence of oxygen. Cells metabolizing glucose by fermentation harvest only about 5% of the available energy in glucose, however. Most organisms use molecular oxygen in a process called cellular respiration. In this series of reactions, the glucose molecule is completely disassembled to yield CO2 and H2O. The process begins with glycolysis; the end product of glycolysis, pyruvate, enters the mitochondrion where it is further metabolized. As far as we know, there are much factors that can influenced the respiration process. That is temperate, light, the water content , the type and age of plant, O2 levels in the air, availability of substrate or other factors. In the plant physiology laboratory will be carrying out a series of experiments that will demonstrate several aspects of respiration including the release of carbon dioxide as a product of respiration and this experiment obtained to known the influence of temperature on the sprout respiration rate.

B. State the Problem From the background above, obtained state the problem :
1. How does the influence of temperature on the sprout respiration rate?

C. Objective From the state the problem above, obtained objective :

1. Known the influence of temperature on the sprout respiration rate.

D. Advantages

From the background above, obtained advantages :

1. In the plant physiology experiments on respiration we can observe and know the

influences that affect the rate of respiration in sprout. In this experiment, factors that affect the speed of respiration is temperature.

CHAPTER II LITERATURE REVIEW

In physiology, respiration (often mistaken with breathing) is defined as the transport of oxygen from the outside air to the cells within tissues, and the transport of carbon dioxide in the opposite direction. This is in contrast to the biochemical definition of respiration, which refers to cellular respiration: the metabolic process by which an organism obtains energy by reacting oxygen with glucose to give water, carbon dioxide and ATP (energy). In plants, respiration occurs in the cell cytoplasm and especially in mitochondria. Following the structure of mitochondria:

Figure 1. Structures of mitochondria. Although physiologic respiration is necessary to sustain cellular respiration and thus life in animals, the processes are distinct: cellular respiration takes place in individual cells of the animal, while physiologic respiration concerns the bulk flow and transport of metabolites between the organism and the external environment. In unicellular organisms, simple diffusion is sufficient for gas exchange: every cell is constantly bathed in the external environment, with only a short distance for gases to flow across. In contrast, complex multicellular animals such as humans have a much greater distance between the environment and their innermost cells, thus, a respiratory system is needed for effective gas exchange.

A. Respiration Cellular respiration, also known as 'oxidative metabolism', is one of the key ways useful cells get energy. This is the set of the metabolic reactions and processes that take place in

organisms' cells to convert biochemical energy from nutrients into adenosine triphosphate (ATP), and then release waste products. Reactions involved in respiration are catabolic reactions that involve the oxidation of one molecule and the reduction of another. Nutrients commonly used by animal cells and plant in respiration include glucose, amino acids and fatty acids, and a common oxidizing agent (electron acceptor) is molecular oxygen (O2). Bacteria and archaea can also lithotrophs and these organisms may respire using a range of inorganic molecules as electron donors and acceptors, such as sulfur, metal ions, methane or hydrogen. Organisms that use oxygen as the final electron acceptor in respiration are described as aerobic, while those that are not referred to as anaerobic. The energy released in respiration is used to synthesize ATP to store this energy. The energy stored in ATP can then be used to drive processes requiring energy, including biosynthesis, movement or transport of molecules across the cell membrane. Judging from his need for oxygen, respiration can be divided into aerobic respiration is respiration that uses oxygen to get energy independent and anaerobic respiration or fermentation process usually called ie respiration does not use oxygen but his material is as carbohydrates, fatty acids, amino acids so that the results respiration in the form of carbon dioxide, water and energy in the form of ATP. Carbohydrates are the main respiratory substrate contained in the cells of higher plants. There are several other important respiratory substrates such as some types of sugars such as glucose, fructose, and sucrose; starch; organic acids, and proteins (used in certain circumstances and species). In general, the respiration of carbohydrates can be written as follows: C6H12O6 + O2 6CO2 + H2O + energy. Reaction equation above is a summary of the reactions that occur in the process of respiration. B. Benefits of respiration Respiration provides many benefits to plants. The benefits can be seen in the process of respiration in which the process of solving organic compounds, the process of solving the dihasilkanlah compounds between important as the "Building Block". Building Block is an important compounds as a body shaper. These compounds include amino acids for protein; nucleotides for nucleic acids, and carbon precursor for profirin pigments (such as chlorophyll and cytochromes), lipids, sterols, carotenoids, flavonoids such as anthocyanin pigments, and certain

other aromatic compounds, such as lignin. It is known that the end result of respiration is CO2 and H2O, this occurs when the substrate is completely oxidized, but when the various compounds in the form, initial substrate respiration was not entirely converted into CO2 and H2O. Only a few substrates respiration completely oxidized to CO2 and H2O, while the rest is used in anabolic processes, particularly in the growing cells. While the captured energy from oxidation perfect some compounds in the process of respiration can be used to synthesize other molecules needed for growth. C. Reaction of respiration In the process of respiration between CO2 produced compounds which are the basis of the process of anabolism. In the process of the fuel cell respiration is hexose sugars. Combustion requires oxygen-free, so that the overall reaction can be written as follows: C 6 h 12 O 6 + 6 CO 2 ------ 6 CO 2 + 6H 2 O + 675 cal In aerobic respiration. Hexose sugars undergo demolition with a very long process. First glucose as a base material having fosfolarisasi, namely the addition of phosphate to the molecule a molecule of glucose to fructose -1, 6 - diphosphate. On phosphorylation, ATP and ADP memgang role as phosphate filler. The conversion of fructose - 1, 6 - dipospat and finally to CO2 and H2O can be divided into four stages, namely glycolysis, the reaction between (oxidative decarboxylation), Krebs cycle, and electron transfer.

Figure 2. Respiration process D. Aerobic Respiration 1. Glycolysis

Glycolysis is a metabolic pathway that is found in the cytoplasm of cells in all living organisms and anaerobic (ie, oxygen is not required). The process converts one molecule of glucose into two molecules of pyruvate, and makes energy in the form of two net molecules of ATP. Four molecules of ATP per glucose produced real, but the two are consumed for the preparatory phase. Initial phosphorylation of glucose is required to destabilize the molecule for cleavage into two triose sugars. During the pay-off phase of glycolysis, four phosphate groups are transferred to ADP by substrate level phosphorylation to make four ATP, and two NADH are produced when the triose sugars oxidized. The whole reaction can be expressed in this way: Glucose + 2NAD + + 2Pi + 2ADP 2pyruvate + 2NADH + 2ATP + 2H + + 2H2O

Figure 3. Glycolysis mechanisms

2. Oxidative decarboxylation of pyruvate Pyruvate is oxidized to acetyl-CoA and CO2 by pyruvate dehydrogenase complex, a

group of enzymes-many copies of each of the three enzymes located in the mitochondria of eukaryotic cells and in the cytosol of prokaryotes. In the process one molecule of NADH is formed per pyruvate oxidized, and 3 moles of ATP formed for each mole of pyruvate. This step is also known as the link reaction, such as links glycolysis and the Krebs cycle. 3. Citric Acid Cycle (Krebs cycle) Citric Acid Cycle also called the Krebs cycle or tricarboxylic acid cycle. When oxygen is present, acetyl-CoA generated from pyruvate molecules created from glycolysis. Once acetyl-CoA is formed, two processes can occur, aerobic or anaerobic respiration. When oxygen is present, the mitochondria will undergo aerobic respiration which leads to the Krebs cycle. However, if oxygen is not present, fermentation of the pyruvate molecule will occur. In the presence of oxygen, when acetyl-CoA is produced, the molecule then enters the citric acid cycle (Krebs cycle) inside the mitochondrial matrix, and will be oxidized to CO2, while at the same time reducing NAD to NADH. NADH can be used by the electron transport chain to create further ATP as part of oxidative phosphorylation. Fully oxidize the equivalent of one glucose molecule, two acetyl-CoA must be metabolized by the Krebs cycle. Two waste products, H2O and CO2, created during this cycle. Citric acid cycle is an 8-step process that involves the enzyme from 8. Throughout the cycle, acetyl-CoA will turn into citrate, isositrat, -ketoglutarate, succinyl-CoA, succinate, Fumarate, malate, and finally, oxaloacetate. Obtain clean energy from one cycle is 3 NADH, 1 FADH, and 1 ATP. Thus, the total amount of energy the entire proceeds from one molecule of glucose (2 pyruvate molecules) is 6 NADH, 2 FADH, and 2 ATP. 4. Electron transport system In the electron transport system lasted packing energy from glucose to ATP. This reaction occurs in the mitochondria membrane, hydrogen from the Krebs cycle is incorporated in FADH2 and NADH converted into electron and protons. In this electron transport system, oxygen is the last electron acceptor, after receiving electrons, O2 reacts with H + to form H2 O. This system produced 34 ATP. E. Anaerobic Respiration If there is no oxygen, the cells do not possess the alternative electron acceptor to produce ATP, so forced electrons derived from glycolysis is transported by organic compounds, a process

called fermentation.

Figure 4. Fermentation of pyruvate to form ethanol or lactic acid Alcoholic fermentation by yeast group by releasing CO2 from pyruvate through decarboxylation and produces two molecules of carbon, acetaldehyde. Acetaldehyde then accept electrons from NADH so that turned into ethanol. Alcoholic fermentation carried out by plants. Lactic acid fermentation by animal cells by transferring electrons from NADH back to pyruvate to produce lactic acid that causes fatigue. F. Factors Affecting Enzyme Activity Regarding these factors can be distinguished, namely: 1. Factor in the cell itself (Internal). Factors affecting respiration in cells there are 4 kinds of, among others, the number in the cell plasma. Tissues of young meristematis where s e l-cells are filled with plasmas typically have a respiratory rate greater than the networks where the number of older plasma is less. Various kinds and number of respiratory enzymes present in the plasma. Number subtarat respiration in cells. 2. Factors outside the cell (External).
a) Temperature.

In general, within the increase of temperature rise also increases respiration rate. In this case, the time factor has a great influence. On the influence of the time factor that causes a reduction in the effective temperature increases respiration rate yet known with certainty. But this is the suspect for several reasons which include, inactivate the enzyme. At high

temperatures the cells can not get enough air to be able to maintain the pace of respiration. At high temperatures the possibility of accumulation of CO2 in the cells to some degree inhibit respiration rate further. When the temperature drops to below 6oC then the reaction rate will decrease until it stops. This drop in temperature among others, will also cause the enzymes occur inactive.
b) Availability of substrate.

Respiration rate of reaction would depend on the availability of substrate, ie compounds that will be parsed through separate chain. Tumbuha n fructans have a reserve starch and lower sugar content will show a low rate of respiration as well. If starvasi (deficiency reserve food) in plants severe, it can also be oxidized proteins. These proteins in hidrolosis into its constituent amino acids, which are then explained to the reactions of glycolytic and Krebs cycle. When the leaves begin to turn yellow, then most of the protein and nitrogen-containing compounds will decompose to chloroplasts. Ammonium ions are freed from the decomposition will be used in the synthesis of glutamine and asparagine. This will prevent plants from ammonium toxicity.
c) O2 levels in the air.

O2 levels in the atmosphere influence the respiration rate will vary depending on the Kinds of networks. But even so the higher levels of O2 in the atmosphere, the higher the speed respirasinya. But keep in mind that most of the variation of atmospheric O2 levels are too small to have a significant effect on the respiration rate. Usually if the changes in levels of atmospheric O2 is less than 5% of atmospheric O2 levels are usually the influence of respiration rate is small (can be ignored).
d) The type and age of plant.

Because of morphological differences between the various types of plants, then there is also the difference between plant respiration rate. Meristematic tissue also showed a higher respiration rate than older plants. Age of plants will affect the rate respirasinya. High respiration rate during germination and remained high at early vegetative growth pase (where the growth rate is high) and then falls with increasing plant age.
e) The water content

In general, with increasing water content in tissue respiration rate will also increase. It

seems clear that the seeds germinated

f) Light

The effect of light on the respiration rate is also generally indirectly. In networks that chlorophyll of plant the light can improve respiration. This is because the light effect on the availability of substrate respiration resulting from the process of photosynthesis. In addition, due to the temperature of the light organs would mennigkat than otherwise because of radiation. The rise in temperature will affect the speed of respiration.
g) Injuries

There his injuries at the plant tissues can cause increased respiration. This is because the sugar levels in the near surface of the piece will increase rather than in cells far away into so this means it will add a lot more respiratory substrate so it can go faster.
h) Mechanical influences

Mechanical actions required in the organs of plants such as by bending or wiggle the organs of plants could increase the speed of respiration in that organ. H intervening acts mechanically on the respiration rate is mainly on aerobic respiration. The increase in respiration rate this sometimes can be up to 100% or more.

CHAPTER III EXPERIMENTAL METHOD A. Kinds of Research This research using experimental method, because there are several variables in it, such as manipulated variables, control variables and the response variable. In this experiment, that will examined by research is the influence of temperature on the sprout respiration rate.

B. Variable : Manipulated variable : Temerature. Response variable Control variable

: The speed, amount of CO2 that released :- Weight of plant (been sprout) -

Kind of plant (been sprout) Volume concentration of NaOH, BaCl2 Volume of HCl titration. Materials : - Been sprout - NaOH solution 0,5 M and HCL 0,5 N - BaCl2 Solution 0,5 N - Phenolftalin solution (PP)

C. Tools and Materials:

Tools : - Erlenmeyer 250 mL (6 pieces ). - Plastic wrapper - Rope - Gauze - Balance scales - Incubator - Pippete
- Buret D. Procedure :

1. Prepared tools and materials that needed in this experiment. 2. Prepare 6 pieces erlenmayer then filled each erlenmeyer with 30 mL of 0.5 M NaOH

solution.
3. Weighing 5 grams of sprouts where provided then wrap with gauze and tied with a

rope. Two samples place in the room temperature and 2 samples for the temperature inside the incubator.
4. Inserting into the erlenmayer and draped the sprouts above NaOH solution with the

help of another rope, then sealed the bottles with plastic.

5. Saving 2 bottles containing sprouts and 1 bottle without bean sprouts (control)

respectively each bottles placed in the placed that has room temperature, the other in an incubator that the temperature is 37 C.
6. Doing titration after 24 hours to determine the amount of CO2 that released during

sprouts respiration.

7. Taking 5 mL of NaOH solution inside the bottle, put in erlenmayer. Then add 2.5 mL

of BaCl2 and drops with to 2 drops of PP so that the solution is red. Furthermore titrating the solution with HCl 0.5 N. Stop the titration after red colour right away.

CHAPTER IV RESULT, ANALYSIS, DISCUSSION A. Observation Result Influence of temperature on the sprout respiration rate can be seen in this table. Table 1. The influence of temperature on the sprout respiration rate. Volume of CO2 Temperature Treatment Volume of HCL (mL) that released (mL) Average

28oC (Room

Control there isnt sprout (A) There is sprout (B)

2,1 1 1 2,3 0,9 0,6

17,4 24 24 16,2 24,6 26,4

17,4 24 24 16,2 25,5 25,5

Temperature) There is sprout (C) Control there isnt sprout (A) 37oC There is sprout (B) (Inkubator) There is sprout (C)

The way to calculate of CO2 that released by using this calculation 5 mL NaOH Volume HCl Titration Volume : N N x 80 : mL 5

Graph 1. Histogram of the influence of temperature on the sprout respiration rate.

B. Analysis

Based on the result observation graph or histogram, it shows that there is different respirationrate between Erlenmeyer that shows as contolled and the Erlenmeyer that filled by mung been sprout at a temperature of 37oC inside the incubator. At first as a control erlenmeyer that contained 0.5 M NaOH(colourless) obtained to white turbid solution (+) when added with 0.5 M BaCl2 (no color), after adding phenolftelin (PP) are obtained colored solution turbid red (+), that colour be able to change into the pink solution then became colorless solution by conduct titration of 2,3 mL HCl 0.5 M (no color). Addition of HCl indicate the number of CO2 bound by NaOH. The volume of CO2 that released in that erlenmeyer is 16,2 mL.

In the second erlenmeyer where placed in the incubator there are sprouts that gained above 0.5 M NaOH solution (colorless), when added by 0.5 M BaCl2(no color) the colour change to be white turbid solution (+ +), then after adding phenolftelin (PP) are obtained a colour solution cloudy pink (+ +), that colour be able to change into the pink solution then became colorless solution by conduct titration of 0,9 mL HCl 0.5 M (no color). Addition of HCl indicate the number of CO2 bound by NaOH. The volume of CO2 that released in that erlenmeyer is 24,6 mL. In the third erlemeyer where placed in the incubator there are sprouts that gained above 0.5 M NaOH solution (colorless), when added by 0.5 M BaCl2 added (no color) the colour change to be white turbid solution (+ +), then after adding phenolftelin (PP) are obtained a solution cloudy pink (+ +), that colour be able to change into the pink solution then became colorless solution by conduvt titration of 0,6 mL HCl 0.5 M (no color). Addition of HCl indicate the number of CO2 bound by NaOH. The volume of CO2 that released in that erlenmeyer is 26,4 mL. On the erlenmeyer where placed in the room temperature (28oC) at first as a control erlenmeyer that contained 0.5 M NaOH(colourless) obtained to white turbid solution (+) when added with 0.5 M BaCl2 (no color), after adding phenolftelin (PP) are obtained colored solution pink turbid (+), that colour be able became colorless solution by conduct titration of 2,1 mL HCl 0.5 M (no color). Addition of HCl may indicate the number of CO2 bound by NaOH. Addition of HCl indicate the number of CO2 bound by NaOH. The volume of CO2 that released in that erlenmeyer is 17,4 mL. At the second erlenmeyer where placed in the room temperature there are sprouts that gained above 0.5 M NaOH solution (colorless), when added by 0.5 M BaCl 2(no color) the colour change to be white turbid solution (+ +), then after adding phenolftelin (PP) are obtained a colour solution pink turbid (+ +), that colour be able to change into the pink solution then became colorless solution by conduct titration of 1 mL HCl 0.5 M (no color). Addition of HCl indicate the number of CO2 bound by NaOH. The volume of CO2 that released in that erlenmeyer is 24 mL. In the third erlemeyer where placed in the room temperature there are sprouts that gained above 0.5 M NaOH solution (colorless), when added by 0.5 M BaCl2 added (no color) the colour change to be white turbid solution (+ +), then after adding phenolftelin (PP) are

obtained a solution pink turbid (+ +), that colour be able to change into the pink solution then became colorless solution by conduvt titration of 1 mL HCl 0.5 M (no color). Addition of HCl indicate the number of CO2 bound by NaOH. The volume of CO2 that released in that erlenmeyer is 24 mL. The amount of CO2 that released in erlenmeyer C same with in erlenmeyer B where placed in the room temperature . C. Discussion Based on the analysis above, it can be seen that temperature affects the amount of CO 2 released from respiration process sprouts, where the incubator temperature (370C) the results obtained by the volume of CO2 respiration is greater than at room temperature. This is because in the incubator temperature, the temperature is kept constant state (stable), where the temperature constant (stable) the enzyme would be optimal without damage. As we know that the process of respiration involves the action of various enzymes. Because the enzyme does not damage the enzyme will accelerate the conversion of glucose to carbon dioxide. Therefore, the CO2 released from sprouts respiration larger. In addition, at higher temperatures the volume of CO2 will be more bound by NaOH so that the levels of CO2 are released bigger. Sprouts were placed in gauze hanging in Erlenmeyer which there NaOH. NaOH solution serves to bind the CO2 gas which produced by the process of respiration. Then a solution of NaOH is added in BaCl2, it will caused the turning solution to be cloudy white colored solution, the more turbid the solution, in the higher respiration process. Then when add by phenolftelin (PP), and doing titration with HCl solution until the pink muddy color was turns into a colorless, it serves to determine amount of CO2 were released. The chemical reaction that takes place is as follows: When NaOH taken from erlenmeyer get the reaction bellow

When NOH added with BaCl2 get the reaction bellow

When added by PP solution and conduct titration using HCl get the reaction bellow

Although NaOH binding CO2 which respiration results. But not at all CO2 could be bound by NaOH. NaOH is which not binding CO2 is cant be reacted with BaCl2 and produce Ba(OH)2 which colorless. Then Ba(OH)2 was tested with PP solution, changing color into the red colour. The red color indicates that the Ba(OH)2 alkaline. When Ba(OH)2 5 ml was titrated by HCl will produces BaCl2 with changing colour indication which originally Ba(OH)2 is red turned into a translucent red (red color right is lost). At the time of the red color that's missing right can be calculated as much as the volume of HCl required to penetrate Ba(OH)2. The volume of HCl is proportional to the volume of NaOH which not binding CO2, so the volume of NaOH that can binding CO2can be known by the volume of HCL. At room temperature (280C) the volume of CO2 sprouts respiration results lower than in incubation temperature (370C). This is because in the lower temperature, the enzyme reaction was not optimal, resulting in the conversion of glucose to CO2 more slowly so that the volume of CO2 released from respiration process respiration process smaller. Controls in this experiment is the only Erlenmeyer filled with NaOH without sprouts, it shows a lower value of respiration. In Erlenmeyer without sprouts is suspected microorganisms perform respiration, because during practice all the tools that are used are not sterilized. Another reason why there is sprouts respiration in NaOH where filled by sprouts faster and produced more CO2 than the NaOH which not filled by sprouts, this is because the respiration is also affected by the substrate for oxidation in the metabolism respiratoris. Generally the substrate for respiration is the substance buried in the relative amounts of many metabolic processes and involves a series of enzymatic reactions that involve the enzyme, the rate of respiration in the existing Erlenmeyer also affected by the enzymes contained in the sprouts and the enzyme will increase when the temperature is high but if the temperature is too high will also damage the enzyme. While Erlenmeyer tubes containing only NaOH that respiration is slow and only produced just a little CO2. This is because there arent enzymatic process there. Based on the analysis above, the experiments on the plant physiology laboratory of just a little. In addition, at lower temperature, the volume of CO2 will be less bound by NaOH so that the CO2 released from

plant respiration can be discussed, the sprouts are used in this experiment from mung bean sprouts 2 days old, this is because the sprouts are as young as 2 days is still active in metabolism, so the energy that get can be used in the growth process. In addition, the sprouts at the age of 2 days has cotyledons which the food reserve storage form of carbohydrates. Carbohydrates are then used for the process of respiration, so most of the carbohydrate is lost as long as the respiration process.

CHAPTER V CLOSING A. Conclussion Based on the experiment above, obtained conclussions that
Temperature affects the rate of respiration At higher temperatures, respiration rate will run faster. But at smaller temperatures,

respiration rate will not run faster. Because in respiration there is enzymatic reaction that involves many enzymes work, then there is an optimum temperature so that the enzyme works optimally. Above the optimum temperature respiration rate will not run faster because the enzyme is damaged. Respiration on germination occurs faster at higher temperatures. The more

CO2 is released, the faster the process of respiration.

BIBLIOGRAPH Bennett, T. P., and Frieden, E.: Modern Topics in Biochemistry, pg. 43-45, Macmillan, London (1969). Campbell, Neil A.; Brad Williamson; Robin J. Heyden (2006). Biology: Exploring Life. Boston, Massachusetts: Pearson Prentice Hall. ISBN 0-13-250882-6. Martinek, R.: Practical Clinical Enzymology: J. Am. Med. Tech., 31, 162 (1969). Salisbury, F. B. & Ross, C. W. 1992. Plant Physiology. Wadsworth Publishing co, California.

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