DEPARTMENT OF BIOLOGICAL SCIENCES

College of Science
University of Santo Tomas
España, Manila 1008, Philippines

Experiment 1

CELL WATER POTENTIAL

The tendency of water to move into or within a system, such as a plant or animal
tissue, the soil or the atmosphere can be measured as the amount of energy per unit
volume (or pressure). Such values, expressed in units of bars or megapascals (MPa) is
referred to as the water potential of the cell and may be computed mathematically as:

Equation (1 –1):  =  s + p + m

where:  = water potential of a cell;  = 0 at equilibrium;

s = solute or osmotic potential; always negative in sign;
p = pressure potential; turgor pressure; may be (-), 0, or (+);
m = matric potential; due to water-binding colloids in the cell; negative in
sign.
Plant cells usually have a negative water potential when they are in equilibrium with
pure water (turgid). Dissolved solutes contribute to the water potential.
The objective of this experiment is to measure the water potential and osmotic
potential in a tissue, then calculate the pressure potential. For practical reasons, a
plant tissue, instead of animal tissue, is used in this experiment.

Materials:
Potato tubers (Large)
Sucrose solutions (0.1m; 0.2m; 0.3m; 0.4m; 0.5m; 0.6m; 0.7m)
Distilled water
No. 5 cork borer
Balances and weighing paper
Plastic cups
Basin
Salt
Funnel, 150mm
Erlenmeyer flasks, 250 ml
Paper towels/filter paper
Blender
Heidenhain (freezing) thermometer / Radioshack Infrared Thermometer
Magnetic stirrers with stirring bars
Potato peelers
Parafilm
Knife
Ruler
Cheesecloth
 DEPARTMENT OF BIOLOGICAL SCIENCES
College of Science
University of Santo Tomas
España, Manila 1008, Philippines

Part I. Determination of Water Potential, 

When a tissue is placed in a sugar solution, there will be a net movement of water
into or out of the tissue depending on the relative water potentials: into the tissue with a
hypotonic solution; out of the tissue in a hypertonic solution; or no net movement in an
isotonic solution when the tissue and solution are at equilibrium. If a series of solutions
of different concentrations of sucrose is used, the sucrose concentration that would
cause no change in weight may be determined by plotting the percent change in weight
against sucrose concentration. Such solution may be assumed to have a water
potential equal to that of the tissue.
Procedure:
1. Prepare eight (8) different sucrose concentrations (dH2O, 0.1m, 0.2m, 0.3m, 0.4m,
0.5m, 0.6m, 0.7m) and dispense 75ml of each separately into 150ml beakers /
plastic cups.
2. Bore 16 cylinders from a large potato using a No. 5 cork borer. Place cylinders in
covered beakers to prevent them from drying out.
3. Cut each cylinder into approximately 4 cm slices. Save remaining potato for Part II.
4. Blot cylinders with paper towels and weigh (to 0.01g) in sets of two. Record the
weight.
5. Put one set of cylinders in each of the beakers with sucrose solutions prepared in
Step 1.
6. After 1.5hrs, remove the cylinders, blot with paper towels and weigh again. Record
results as before.
7. Compute for the change in weight (W) and the percent change in weight (%W)
using the following formulae.
Equation (1–2): W  W f  Wi

W
Equation (1–3): % W  x 100
Wi
where: Wi = initial weight of the cylinders;
Wf = final weight of the cylinders.
8. Tabulate all results. Plot % change in weight vs. sucrose concentration. Draw the
best-fit straight line through the points.

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 DEPARTMENT OF BIOLOGICAL SCIENCES
College of Science
University of Santo Tomas
España, Manila 1008, Philippines

9. Determine molal concentration of sucrose that gives 0% change in weight.

Compute for s in bars of that sucrose solution using the formula:
Equation (1–4): s = -miRT
where: m = molality; (NOTE: 1 molal = 1 x 103 mol m-3 H2O)
i = ionization constant = 1 for sucrose
R = gas constant = 8.31 J K-1 mol-1
T = room temperature in K (oC + 273)

10. Determine the water potential of the potato assuming that m is very small and
therefore negligible. Convert unit of J m-3 (energy per unit volume, pressure which
is equivalent to Pa) to MPa by dividing computed value by 106.
NOTE: In a free standing solution, p = 0; so  = s.

Part II. Determination of solute potential (s) of extracted sap by cryoscopy

Cryoscopy (cry·os·co·py) is the examination of liquids, based on the principle
that the freezing point of solutions varies according to the amount and the nature of the
solute. This provides a relatively easy means of arriving at the osmotic potential of a
solution. The freezing point of an aqueous solution decreases with an increase in
dissolved solutes. Once the freezing point of a solution has been determined, its solute
potential may be calculated, thus:

Equation (1–5): s = 1.22Tƒ

where: s = solute potential in MPa at 0oC
Tƒ = freezing point of the solution in oC
The equation holds true for freezing temperature. The solute potential of the
extracted sap at room temperature can then be computed:
Equation (1–6): s = 1.22Tƒc
where: c is the correction factor; (Rm temp in K/273K)

Procedure:
Sap extraction
11. Peel 2 – 4 potatoes, chop and place in a blender. Puree the tissue.
12. Filter blended potato with cheesecloth to remove cell wall and debris. Store in
covered beaker / plastic cup.
Freezing point determination
13. Immerse heidenhain thermometer in crushed ice – salt bath. Record reading.

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 DEPARTMENT OF BIOLOGICAL SCIENCES
College of Science
University of Santo Tomas
España, Manila 1008, Philippines

14. Pour 60ml sap in 250ml Erlenmeyer flask with magnetic stirring bar and insert
thermometer. Surround flask with ice–salt bath (see Figure 1–1). Stir vigorously.
15. When temperature reads about 1oC, read temperature every 10 sec.
NOTE: There should be a continuous drop in temperature, followed by a short
plateau which corresponds to the freezing point, is followed by an increase
16. Plot temperature vs. time.

Heidenhain thermometer

Erlenmeyer flask Rubber stopper

Beaker

Ice-salt bath
Stir bar
Sap extract
Magnetic stirrer 4
5 6
7 4
5 6
7

3 8 3 8

2 9 2 9
1 11 1 10

Figure 1 – 1. Setup for freezing point determination

17. Determine the true freezing point, Tƒ.

Equation (1–7): Tƒ = Tƒ’ – 0.0125ts
where: Tƒ’ = apparent freezing point
ts = degrees of supercooling; (-) in sign
ts = lowest temp - Tƒ’
0.0125 = amount of water (1/80) solidifying per degree of
supercooling
18. Make another correction for the zero point of the thermometer.
19. Determine the solute potential using Equation (1–5) at 0oC.
20. Calculate the pressure potential (p) of the cells of potato.

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