Cell Water Potential Experiment 1

Submitted by: Venturina, Russell Evan L. Aragoza, Carlos Dominic P. Olanka, Nicanor Relova, Mikhail Angelo
Submitted to: Prof/s: Rey Donne Papa & Mae Lowe Diesmos Date Submitted: December 7, 2011

I.

Introduction

Water is essential for survival. It covers 70.9% of the Earth's surface,and is vital for all known life forms. It is responsible for structural support through turgor pressure in plants. The tendency of water to move across a gradient with a higher concentration is called water potential. Osmosis is the diffusion of water across a semi-permeable membrane. The presence of a solute decreases the water potential of a substance. In a cell, which has so many organelles and other large molecules, the water flow is typically into the cell.Hypertonic solutions are that which contains more solute (lower water potential) is present. Hypotonic solutions are those with less solute (higher water potential). Isotonic solutions have equal concentrations of substances. Water potentials are thus equal, although there will still be equal amounts of water movement in and out of the cell, the net flow is zero. In the process of osmosis, water moves from an area of higher energy to an area of lower energy. Water potential is the measure of free energy of water in a solution. Water potential ( )is based on s, p, and m, and can be computed mathematically by the formula: = s + p + m Water potential ( ) is pressure/unit volume, and is the measure of the tendency of water to move from a system to another and is expressed bymegapascals(MPa). When it is zero, then it is in equilibrium. Solute potential, also called osmotic potential, ( s) characterizes the effects contributed by the dissolved solutes in water; pressure potential ( p) represents the effects contributed by turgor pressure, whereas matric potential ( m) signifies the effects contributed by water-binding colloids in the cells. s is always negative in sign; p is either positive or negative, while m is also negative. Potatoes (Solanumtuberosum) are starchy edible tuber native to South America and cultivated all over the world. The tuber, the edible part of the white potato is short and thick, starchystem; with the "eyes" (or the dark spots) actually are the buds on the stem. In this experiment, we are to measure the water potential and osmotic potential in a tissue of the plant sample Solanum tuberosum, then calculate the pressure potential.

II.

Materials and Methods

Materials: y Potato Tubers y Sucrose (C12H22O11) solutions (0.1 m; y 0.2m; 0.3m; 0.4m; 0.5m; 0.6m; 0.7m) y Distilled water y No.5 Cork borer y Balances and weighing paper y Plastic cups y Funnel, 150 mm y Paper towels y Blender y Heidenhain (freezing) thermometers y Potato Peelers y Knife

y y

Ruler Cheesecloth

Methods Part 1: Determination of Water Potential, Eight different sucrose concentrations (Distilled water, 0.1m, 0.2m, 0.3m, 0.4m, 0.5m, 0.6m, 0.7m) were prepared and were dispensed separately into plastic cups. Using the cork borer, 16 cylinders were bore from a large potato and were cut into 4 cm slices. The cylinders were blotted with paper towels and were weighed in sets of two. The weight was recorded as the initial weight (Wi). One set of cylinders were placed in each of the plastic cups containing the sucrose solutions. After 30 minutes, the cylinders were removed, blotted with paper towels, and were weighed again. The weight was recorded as the final weight (Wf). The change in weight ( W) and the percent change in weight (% W) were computed using the following formula: W = Wf - Wi % W= x100 All results were tabulated. The % change in weight was plotted against sucrose concentration, the best-fit line drawn through the points. The molal concentration of sucrose that gives 0% change in weight was determined. The sin bars of that sucrose solution was computed using the following formula: s = -miRT In this equation, m is the molar concentration of sucrose that gives 0% change in weight, i is the ionization constant (1 for sucrose), R is the gas constant (equal to 8.31 J K-1 mol-1), and T is the room temperature expressed in Kelvins (C + 273). The water potential of the potato was determined under the assumption that the value of mis small and therefore negligible.

Part 2: Determination of solute potential sof extracted sap by cryoscopy The leftover potatoes were chopped and pureed in a blender. The blended potato was filtered using cheesecloth to remove cell wall and debris; its filtrate was placed in a beaker. The temperature of the crushed ice-salt bath was obtained by immersing the heidenhain thermometer. The reading was recorded. 60 mL of the sap was placed in a beaker. The thermometer was inserted into the beaker and was placed into the ice-salt bath, stirring vigorously. When the temperature read 1C, the temperature was read and recorded every 10 seconds. A plot of the temperature versus time was made.

III.

Results and Discussion Initial Weight in Grams (Wi) Final Weight in Grams (Wf) Change in Weight (W=Wf-Wi) 0.7919 0.0068 -0.0441 -0.2140 -0.5274 -0.7712 -0.8690 -1.1274
%Change in Weight

Sucrose Concentration

Dist. H2O 7.0412g 7.8061g 0.1m 7.0139g 7.0207g 0.2m 7.0709g 7.0268g 0.3m 6.9455g 6.7315g 0.4m 6.7212g 6.1938g 0.5m 7.0574g 6.2862g 0.6m 6.8763g 6.0073g 0.7m 7.1182g 5.9908g Table 1. Results of Determination of Water Potential

11.2 0.096 -0.624 -3.081 -7.847 -10.9275 -12.6376 -17.8977

In the cups containing distilled water 0.1, and 0.2 molal sucrose solutions, the potatoes increased in weight because the potatoes absorbed water due to the potatos lower water potential with respect to the solution. Therefore, the percent change in weight is positive. In the cups containing 0.3 - 0.7 molal sucrose solution, the potatoes decreased in weight because water left due to the potatos higher water potential with respect to the solution. Therefore, the percent change in mass is negative.

___________________________________ Graph 1. Determination of Water Potential

Temperature (Celsius) 19.7 17.3 15.2 14.8 3.8 -3.6 -4.6 -1.4 -0.8 -0.3 -0.2 -0.2 -0.2

Time (seconds) 60s 120s 180s 240s 300s 310s 320s 330s 340s 350s 360s 370s 380s

-0.2 390s Table 2. Determination of Solute Potential of extracted sap through cryoscopy In a typical freezing point curve, there is a continuous drop in temperature as the liquid is cooled, then it remains constant (plateau) signifying that both liquid and solid phases are present. Temperature finally drops again when only solid phase is present. From the temperature vs. time curve shown below, the temperature became stable for 10 seconds at 0.2 C. This will be the designated freezing point for the sap extract.

Graph 2. Determination of Solute Potential of extracted sap through cryoscopy

However, this is merely an estimate and is called the apparent freezing point. The true freezing point (Tf) is obtained after correcting for super-cooling according to the following equation: Tf = Tf' - 0.0125 ts ts = Lowest temperature - Tf' Where Tf is the true freezing point and Tf' is the apparent freezing point. ts is the degrees of super cooling (negative in sign), while 0.0125 corresponds to the amount of water (1/80) that solidifies per degree of super cooling. Tf = Tf' - 0.0125 ts = -0.2 0.0125 (-0.3- -0.2) = -0.2 (-0.00125) = -0.20125 -0.2 Correction was made for the zero point of the thermometer to determine the solute potential. Tf = Tf' - 0.0125 ts = 0 0.0125 (-0.2 0) = -0.0025 The solute potential at 0C can now be determined: s = (1.22 MPa deg-1) Tf x (temp in K/273 K) = (1.22) (-0.2) C x [(0+273)/273 K] = -0.244 MPa At room temperature however, s = (1.22 MPa deg-1) Tf x (temp in K/273 K) = (1.22) (-0.2) C x [(25+273)/273 K] = -0.2663 MPa Finally, the pressure potential ( p) of the cells of the potato can be solved by the formula for water potential: = s + p ( m is neglected) Under equilibrium conditions, the water potential of the potato is equal to the water potential of the 0.23 molal sucrose solutions. Therefore, the water potential ( ) of the potato is -0.57 MPa. The computed solute potential of the potato sap extract at room temperature is -0.0025 MPa. = s+ p -0.57 MPa = -0.0025 MPa + p p = 0.0025 MPa - -0.57 MPa p = .5725 MPa

Sources: Purves et al. Life: The Science of Biology 4th Edition. USA: Sinauer Associates &WH Freeman Salisbury, F. B. and C. W. Ross. 1992. Plant Physiology, 4th Ed. Wadsworth Publishing Co., Belmont, CA. Saupe, S. G. (2009, January 7). Determining Osmotic Potential by the Freezing Point Depression Method. Retrieved December 6, 2011, from Plant Physiology (Biology 327) Home Page: http://employees.csbsju.edu/SSAUPE/biol327/Lab/water/water-labfreez.htm