Measurement of Osmotic Potential ( ) by Incipisnt Plasmolysis

GROUP III Yunandar/1114040181 Surhayanti Amir/1114040192 Sri Vianita/1114040195 Sri Wahyuningsih/1114040199

BIOLOGY ICP

UNIVERSITAS NEGERI MAKASSAR TAHUN AJAR 2012/2013

CHAPTER I INTRODUCTION
A. Background Water potential (w, psi), which is a measure of the energy state of water is affected by dissolved solutes, pressure and matrix particles. The contribution to water potential by dissolved solutes, termed osmotic potential (s ), is always negative in sign. In other words, solutes decrease the water potential. The contribution of pressure (p) may be positive, negative or zero, but is generally positive since most plant cells are turgid (turgor pressure). The contribution due to the binding of water to colloidal particles (matric) and surfaces, termed matric potential (m), also lowers the water potential. Although it is often small enough to be ignored, matrix potential is important when considering soil water relations. Thus, the water potential of a plant system can be arithmetically represented by the equation: w = s + p + m B. Purpose Measurement of osmotic potential by incipisnt plasmolysis

CHAPTER II BASIC THEORY

Now, if we were to take two containers of water, and separate them by a semi-permeable membrane (one that allows water to go through its pores, but not solutes like salt or sugar), and add sugar to one side, this would result in a lowering of the kinetic energy of the water-sugar solution. Thus, from a statisticalprobability point of visw, we would expect the molecules of pure water to encounter the membrane more often than the lower energy water molecules on the solution side, and thus, over time, water will move from the pure water to the solution. Of course some water molecules do go the other way, but the net exchange favors movement into the solution. This is known as osmosis. It is a special case of diffusion (Anonymous, 2013). If a cell is placed in a solution which has a that is higher than that of the cell, there will be a net movement of water into the cell. However, if the surrounding solution has a lower than in the cell, there will be a net movement of water out of the cell. If this latter situation continues, the plasma membrane and cytoplasm will pull away from the cell wall, a condition known as 3 plasmolysis. By trial and error, a concentration of bathing solution can be found that just produces plasmolysis, and this is known as incipisnt incipisnt plasmolysis is defined as when plasmolysis. Thus, 50% of the cells are plasmolyzed. At

incipisnt plasmolysis, there is no longer a pressure potential exerted by the wall (i.e., = 0), and therefore, under that condition, = . It should also be noted that for solutions, = . A solution which just causes incipisnt plasmolysis thus has a water potential (and osmotic potential) of the cell cytoplasm. Finally, since the cell we use are highly vacuolated, it can also be assumed that the osmotic potential of the cell is basically the vacuolar osmotic potential (Ismail, dkk. 2013). Thus, the solution will increase in volume, and become more diluted. Over time, this will slow the flux of water into the solution, but not stop it entirely. However, eventually, the weight of the water will exert a backpressure on the solution, which, if given enough time (and large enough container) will increase the pressure on the membrane and force water molecules to go back into the pure

water. If the pressure is great enough, it can totally balance the number coming in, and the net flux of water will cease. The amount of pressure needed to totally balance the flows of water is known as the osmotic pressure and symbolized aswith units of pressure (e.g., pounds per square inch, atmospheres, bars, Megapascals) (Anonymous, 2013). A slightly more complex theory that is often found in general biology books (including your text, p. 117) is the bound water explanation. This says that any hydrophilic solute (like sucrose or NaCl) will bind up hydrating water and prevent it from moving freely. Therefore, the side of a semipermeable membrane with pure water has a higher free water concentration than the side with the solute molecules. According to this explanation, free water moves into hypertonic solutions simply because it is diffusing down its concentration gradisnt. Although it is popular in introductory texts, this theory is not even mentioned in several revisws (Baumgarten and Feher, 1998; Weiss, 1996, pp. 216-222). If the bound water explanation were true, we would expect that a greater mass of hydrophilic solute would bind more water. Whether a certain mass of solute is present in a few large molecules or in many small ones shouldnt matter. Also, when predicting osmosis, we would have to carefully consider how hydrophilic the solute is (that is, how many water molecules it binds per molecule). In fact, the number of molecules present does affect osmosis, and we can predict osmosis without considering how hydrophilic the solute molecules are (Anonymous, 2013). Water is a simple molecule, consisting of one atom of oxygen (0) and two hydrogen atoms (H), so that the molecular weight of only 18 g / mol. In spite of the simplicity of the composition of the constituent atoms and small molecular size, the water molecule has several unique characteristics. These characteristics caused by a seriss of two H atoms on atom 0 (in center) do not form a straight line. This circuit makes an angle of 1050. The magnitude of this angle is always the same if the water is in solid form (ice), but rather variss if water is in liquid

form, although the average angle remains 1050. Water can dissolve more types of chemicals compared to other liquids (Lakitan, Benjamin. 2011). If plant cell are placed in pure water, water will initially move into the cell. After are period of time the cell will become turgid. Turgor pressure is the pressure exerted against the cell wall by contents of the cell. At first most water movement is into the cell. As the turgor pressure increases water will begin to diffuse out of the cell at a greater rate, eventually equilibrium will be reached and water will enter and leave the cell at the same rate. This stage is used to find the water potential of a particular cel (Anonymous, 2013). Intake or water net expenditure by a cell occurs by osmosis, is passive transport of water through a membrane. The combined effect of these two factors solute concentration and pressure are called water potential. In the water potential is important to understand is the water will move through the membrane from a solution with high water potential to a solution with a lower potential IAR. Components potential in water potential refers to the potential energy, which is the capacity to perform work when water moves from areas with higher to areas with lower (Campbell, 2000). Potato cell contain polysaccharides starch and glycogen they are good for storage. The potato cell is surrounded by plasma membrane it is a fluid mosaic model, which is mosaic of phospholipids and proteins moving around they are not solid. This is why plant cell can become turgid and flaccid because their walls (plasma membrane) can stretch. The plasma membrane is a selectively permeabel barrisr between the cell and the extra cellular environment. Water enters in the cell through phospholipids (Anonymous, 2013). Like molecular diffusion and pressure deriven bulk flow, osmosis occurs spontaneously in response to a driving force. In simple, diffusion, substances move down a concentration gradisnt; in pressure-driven bulk flow, substances move down a pressure gradisnt; in osmosis, both types of gradisnts influence transport, he is say the direction and rate of water flow across a membrane are determined not solely by the concentration gradisnt of water or by pressure gradisnt, but by the sum of these two driving (Finkelstein, 1987).

CHAPTER III PRACTICUM METHOD

A. Place and Date Day / date Time Place : Wednesday, March 21st 2013 : 10.50 Wita 12.30 Wita : Biology Laboratory third floor at west FMIPA UNM

B. Tools and Materials 1. Tools a. Microscope b. Cutter c. Object and deck glass d. Petri dish e. Tweezers 2. Material a. Solution of sucrose b. Leaf of Rhoeo discolor C. Work Procedure
1. Prepare 6 petri dish and label each petri dish by concentration sucrose

solution to be used.

2. fill each petri dish with a solution sucrose 0.1, 0:15, 0.20, 0:25, 0:30, 0:40

m.

3. Taking epidermis Rhoe discolor, then slashing or slicing the epidermal

layer purple with a knife or razor blade and slashed seek only the cell layer.

4. Submerge the epidermis slashes on a petri dish that already contains a

certain concentration of sucrose solution with the same number of incisions for 15 minutes.

5. After 15 minutes, take the cuts that have been soaked in a petri dish and

examined under a microscope.

6. Counting the total number of cells in one area of the fisld of vision, the amount of cell is happened plasmolisis and the percentage of cells that happened plasmolisis the total number of cells.

CHAPTER IV RESULT
A. Result of Practicum Table Effect of sucrose concentration on epidermal cells Rhoe discolor

Caption Concentration 0,10 Plasmolisis Happen % 60 % Not Happen Plasmoliss % 50 % Pictures osmotic potential () for 0.1M in 27C : - = miRT - = (0.1)(1)(0.082) (273+27) - = -2.46 = 2.46 osmotic potential () for 0.15 M in 27C : - = miRT - = (0.15)(1)(0.082) (273+27) - = -3.69 = 3.69 osmotic potential () for 0.20M in 27C : - = miRT - = (0.20)(1)(0.082) (273+27) - = -4.96 = 4.96 osmotic potential () for 0.25M in 27C : - = miRT - = (0.25)(1)(0.082) (273+27) - = -6.15 = 6.15 osmotic potential () for 0.30M in 27C : - = miRT - = (0.30)(1)(0.082) (273+27) - = -7.38 = 7.38 osmotic potential () for 0.40M in 27C : - = miRT - = (0.40)(1)(0.082) (273+27) - = -9.84 = 9.84

0,15

20 %

80 %

0,20

40 %

60 %

0,25

70 %

30 %

0,30

80 %

20 %

0,40

80 %

20 %

B. Data Analysis

Based on the data that have been obtained can be analyzed as follows: 1. At a concentration of 0.10 m sucrose solution. Epidermal cells Rhoe discolor experisnced plasmolisis with the percentage of cells is happened plasmolisis by 60% and 50% were not plasmolisis. 2. At a concentration of 0.15 m sucrose solution. Epidermal cells Rhoe discolor experisnced plasmolisis with the percentage of cells is happened plasmolisis by 20% and 80% were not plasmolisis. 3. At a concentration of 0.20 m sucrose solution. Epidermal cells Rhoe discolor experisnced plasmolisis with the percentage of cells is happened plasmolisis by 40% and 60% were not plasmolisis. 4. At a concentration of 0.25 m sucrose solution. Epidermal cells Rhoe discolor experisnced plasmolisis with the percentage of cells is happened plasmolisis by 70% and 30% were not plasmolisis. 5. At a concentration of 0.30 m sucrose solution. Epidermal cells Rhoe discolor experisnced plasmolisis with the percentage of cells is happened plasmolisis by 80% and 20% were not plasmolisis. 6. At a concentration of 0.40 m sucrose solution. Epidermal cells Rhoe discolor experisnced plasmolisis with the percentage of cells is happened plasmolisis by 80% and 20% were not plasmolisis. C. Discussed When Rhoeo discolor under normal circumstances, visible cell parts hexagonshaped cavity with cytoplasm of the cell wall purple meet. Water dripped form an isotonic environment both inside and outside the cell, so that the normal cell shape. At the time of incision Rhoeo discolor leaves, soaked in a solution of sucrose 0:10, 0:15, 0:20, 0:25, 0:30, 0:40 m. So the solution is more concentrated outside the cell than inside the cell. In accordance with the principle of osmosis, the movement of water or solvent from a more dilute solution to a more concentrated

solutions. water will flow out of the cell vacuoles heading out because of the pressure of osmosis. Consequently Rhoeo discolor leaf cells lose water so purple cytoplasm away from the cell walls shrink and as if out and rupture of the cell. Gradually the cytoplasm fade into purple blotches. This happens because the solution sucrosa acts as a hypertonic solution, the solution whose concentration is lower than the fluid inside the cell. From the analysis above, it can be derived that the dense concentration of sucrose solution is used to soak the incision epidermis Rhoe discolor the more the epidermal cells that undergo plasmolisis. This can be the result of differences in water potential inside and outside the cell. Potential water in the cell is greater than the existing water potential outside the cell. Therefore, the water potential is proportional to the osmotic potential, the osmotic potential in the cell is greater than the osmotic potential that exists outside the cell. This has led to the migration of water molecules in the cell to outside the cell in the lab this time the water molecules move from epidermal cells Rhoe discolor leading to the solution of sucrose, resulting protoplasts epidermal cells lose water, shrink volume (cells become wrinkled) and finally detached from the cell wall, the events that occur in epidermal cells Rhoe discolor is commonly called the Plasmolisis.

CHAPTER V CONCLUSION AND SUGGESTION

A. Conclusion Plasmolisis event is the release of the cell membrane in plant cells due to the cell is in an environment that is hypertonic. Conditions hipotonis cells resulting environmental occurrence osmosis from the cells into the environment. As a result, water levels dropped dramatically in the cell and the cell membrane detached from the cell wall.

A cell will undergo plasmolisis if the water potential in the cell greater than the existing water potential outside the cell. It also means that the osmotic potential that is inside the cell is greater than outside the cell. B. Suggestion
1. Laboratory should provide tools that fit the needs of that practice can be

implemented with a conducive and comfortable.

2. Assistant should accompany each group to support the implementation of

practical activitiss in accordance with the desired.

3. My frisnds should understand the working procedures before entering the lab

room

BIBLIOGRAPHY
Anonymous, 2013. http://courseworkbank.info/journal. Accesed 26th march 2013 Anonymous, 2013. Lecture Water. http://employees.csbsju.edu. Accesed 26th march 2013 Anonymous, 2013. http://biology.clemson.edu. Accesed 26th march 2013 Anonymous, 2013. http://appstate.edu. Accesed 26th march 2013. Campbell. 2000. Biologi Campbel edisi 3. Jakarta: Erlangga.

Lakitan, Benyamin. 2011. Dasar-dasar Fisiologi Tumbuhan. Jakarta: Rajawali Pers Finkelestein, A. (1987) Water Movement through Lipid Bilayer, Pores, and Plasma Membranes: Theory and Reality. Wiley, New York. Taiz, Zeiger. 2002. Plant Physiology edtion 3. Sinauer Associates: England

Questions:
1. What concentration of surcose resulted in incipisnt plasmolysis, and how did you know when it occurred? 2. Based on the above, what was the osmotic potential of the cells? Show your calculation. 3. What were possible sources of error in this experiment?

Answer
1. In our observation, of all the sucrose concentration given all the impact

plasmolisis. But the presentation of the different plasmolisis. Experisncing the highest plasmolisis is sucrosa solution 0:30 and 0:40 then sequentially is 0:25, 0:10, 0:20 and presentations that have the lowest plasmolisis is 0.15 m sucrose solution.We known plasmolisis happen becouse the water molecules move from epidermal cells Rhoe discolor leading to the solution of sucrose, resulting protoplasts epidermal cells lose water, shrink volume (cells become wrinkled) and finally detached from the cell wall, the events that occur in epidermal cells Rhoe discolor is commonly called the Plasmolisis. 2. Observation result of with use abbreviation:

osmotic potential () for 0.15 M in 27C : - = miRT - = (0.15)(1)(0.082)(273+27) - = -3.69 = 3.69

osmotic potential () for 0.1M in 27C : - = miRT - = (0.1)(1)(0.082)(273+27) osmotic potential () for 0.30M in 27C : - = -2.46 - = miRT = 2.46 - = (0.30)(1)(0.082)(273+27) - = -7.38 = 7.38

osmotic potential () for 0.40M in 27C : - = miRT osmotic potential () for 0.20M in 27C : - = (0.40)(1)(0.082)(273+27) - = miRT osmotic potential () for 0.25M in 27C : - (0.20)(1)(0.082)(273+27) -==-9.84 miRT == 9.84 -4.96 - (0.25)(1)(0.082)(273+27) = 4.96 - -6.15 = 6.15
3. Errors

that

may

occur in this lab are:

a. Aprentice inaccuracy when determining or calculating the number of

cells undergoing plasmolisis and the number of cells that do not undergo plasmolisis.
b. Errors in taking the epidermis rhoe discolor, possible incision

epidermis has taken bold measures to normal size in the experiment to be performed.
c. Sucrose solution used was not valid due to the mixture of sucrose

solution with each other this is caused by the use of a Pasteur pipette solution simultaneously for all becouse Pasteur pipette is used only one solution for all.